RU2841609C2 - Cd19-targeted chimeric antigen receptors and ways of use thereof in immunotherapy - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to a polypeptide which binds CD19 containing a CD19-directed chimeric antigen receptor (CAR), as well as cells expressing it. Also disclosed is a polynucleotide coding said CD19-directed chimeric antigen receptor (CAR), as well as a method of producing a population of natural killer (NK) cells expressing a CD19-directed chimeric antigen receptor (CAR). Invention also relates to a pharmaceutical composition containing a plurality of natural killer (NK) cells expressing a CD19-targeted chimeric antigen receptor (CAR).

EFFECT: invention is effective for treating a subject with cancer in which CD19 is expressed.

30 cl, 39 dwg, 11 ex

Description

RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application Nos. 62/814,180, filed March 5, 2019, 62/895,910, filed September 4, 2019, and 62/932,165, filed November 7, 2019, the entire contents of each of which are incorporated herein by reference.

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

[0002] Some embodiments of the methods and compositions provided herein relate to CD19-directed receptors. In some embodiments, the receptors are chimeric. Some embodiments include methods of using chimeric receptors in immunotherapy.

PREREQUISITES FOR THE CREATION OF THE INVENTION

[0003] As more is learned about the different types of cancer and the characteristics of a cancer cell that can be used, in particular, to distinguish such a cell from a healthy cell, therapeutic agents are being developed that exploit the distinctive features of the cancer cell. Immunotherapies that use engineered immune cells represent one approach to treating cancers.

INCLUSION OF MATERIAL BY LINK IN THE FORM OF AN ASCII TEXT FILE

[0004] This application incorporates by reference the sequence listing contained in the following ASCII text file, which was filed concurrently with this application. File name: NKT033WO_ST25.txt; created February 8, 2020, size 434 KB.

SUMMARY OF THE INVENTION

[0005] Immunotherapy is a new technological advance in the treatment of disease in which immune cells are engineered to express specific targeting and/or effector molecules that specifically identify and respond to diseased or damaged cells. This represents a promising improvement, due at least in part to the ability to specifically target diseased or damaged cells, as opposed to more traditional approaches such as chemotherapy, where all cells are targeted and the desired outcome is that sufficient healthy cells survive to allow the patient to remain alive. One immunotherapy approach is the recombinant expression of chimeric receptors in immune cells to achieve targeted recognition and destruction of aberrant cells of interest.

[0006] In several embodiments, provided herein is an immune cell, as well as populations of immune cells, that express a CD19-directed chimeric receptor, wherein the chimeric receptor comprises an extracellular CD19 binding moiety, a hinge and/or transmembrane domain, and an intracellular signaling domain. Also provided herein are polynucleotides (as well as vectors for transfecting cells therewith) encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, a hinge and/or transmembrane domain, and an intracellular signaling domain.

[0007] In several embodiments, a polynucleotide encoding a CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) of a single chain variable fragment (scFv) and a light chain variable domain (VL) of an scFv, a hinge, a transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises an OX40 subdomain, a CD3-zeta subdomain.

[0008] In several embodiments, a polynucleotide encoding a CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) of a single chain variable fragment (scFv) and a light chain variable domain (VL) of an scFv, wherein the encoded VH domain comprises at least one heavy chain complementarity determining region (CDR) selected from the group consisting of SEQ ID NO: 133, SEQ ID NO: 134, and SEQ ID NO: 135, wherein the encoded VL domain comprises at least one light chain complementarity determining region (CDR) selected from the group consisting of SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO: 129, a hinge domain, a transmembrane domain, an intracellular signaling domain, and wherein the intracellular signaling domain comprises an OX40 subdomain and a CD3-zeta subdomain.

[0009] In some embodiments, the polynucleotide further encodes membrane-bound interleukin-15 (mbIL15). However, in some embodiments, a separate polynucleotide is used to encode mbIL15. In some embodiments, the transmembrane domain is derived from or comprises a CD8-alpha transmembrane domain. In some embodiments, the CD8-alpha transmembrane domain is encoded by the sequence of SEQ ID NO: 3. In some embodiments, the hinge is derived from or comprises a CD8-alpha hinge. In some embodiments, the CD8 alpha hinge is encoded by the sequence of SEQ ID NO: 1. In some embodiments, the OX40 subdomain is encoded by a sequence having at least 90% (e.g., 90-95%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 5. In some embodiments, the CD3 zeta subdomain is encoded by a sequence having at least 90% (e.g., 90-95%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 7. In some embodiments, mbIL15 is encoded by a sequence having at least 90% (e.g., 90-95%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 11. In several embodiments, the OX40 domain is encoded by the sequence of SEQ ID NO: 5, the CD3-zeta subdomain is encoded by the sequence of SEQ ID NO: 7, and/or mbIL15 (whether encoded separately or bicistronic) is encoded by the sequence of SEQ ID NO: 11. In several embodiments, the encoded OX40 subdomain comprises the amino acid sequence of SEQ ID NO: 6, the encoded CD3-zeta subdomain comprises the amino acid sequence of SEQ ID NO: 8, and/or the encoded mbIL15 (whether encoded separately or bicistronic) comprises the amino acid sequence of SEQ ID NO: 12.

[0010] In some embodiments, the VH domain comprises a VH domain selected from SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123, and wherein the VL domain comprises a VL domain selected from SEQ ID NO: 117, SEQ ID NO: 118, and SEQ ID NO: 119. In some embodiments, the polynucleotide encodes a VL domain comprising at least one light chain complementarity determining region (CDR) selected from the group consisting of SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO: 129. In some embodiments, the polynucleotide encodes a VH domain comprising at least one heavy chain complementarity determining region (CDR) selected from the group consisting of SEQ ID NO: 133, SEQ ID NO: 134, and SEQ ID NO: 135. In several embodiments, the polynucleotide is designed (e.g., engineered) to reduce the potential antigenicity of the encoded protein and/or enhance one or more characteristics of the encoded protein (e.g., target recognition and/or binding characteristics). As a result, in accordance with several embodiments, the CD19 binding fragment does not comprise certain sequences. For example, in several embodiments, the polynucleotide does not encode one or more of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, or SEQ ID NO: 55. In several embodiments, the encoded VH domain comprises an amino acid sequence having at least 90% (e.g., 90-95%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 120. In some embodiments, the encoded VH domain comprises an amino acid sequence of SEQ ID NO: 120. In some embodiments, the encoded VL domain comprises an amino acid sequence having at least 90% (e.g., 90-95%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 118. In some embodiments, the encoded VL domain comprises an amino acid sequence of SEQ ID NO: 118. In some embodiments, the VH domain is derived from a parent amino acid sequence and modified from the parent sequence. For example, mutations, truncations, extensions, conservative substitutions, or other modifications are introduced to increase the affinity of the domain for its target, increase the avidity for the target, and/or decrease the antigenicity potential of the sequence. In some embodiments, the VH domain is derived from a humanization of the VH domain amino acid sequence set forth in SEQ ID NO: 33. Similarly, in some embodiments, the VL domain is derived from a humanization of the VL domain amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the polynucleotide encodes a CD19-targeted chimeric antigen receptor that comprises a sequence having at least 90% (e.g., 90-95%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence set forth in SEQ ID NO: 187. In some embodiments, the polynucleotide encodes a CD19-targeted chimeric antigen receptor comprising the amino acid sequence set forth in SEQ ID NO: 187.

[0011] In several embodiments, the polynucleotide does not encode or otherwise comprise a DAP10 domain. In several embodiments, the polynucleotide does not encode or otherwise comprise a DAP12 domain. In several embodiments, the intracellular signaling domain comprises additional subdomains that advantageously enhance the generation of cytotoxic signals by cells expressing the constructs. In several embodiments, the polynucleotide further encodes one or more of CD44 and CD27 as signaling subdomains. In several embodiments, the polynucleotide optionally further encodes a detection label or other moiety (e.g., a marker) that allows for detection of expression of the protein(s) encoded by the polynucleotide by host cells.

[0012] This document also provides methods for using the disclosed polynucleotides in the manufacture of a medicament for enhancing NK cell cytotoxicity in a mammal in need thereof, in the manufacture of a medicament for the treatment of cancer in a mammal in need thereof, and/or in the treatment of cancer in a mammal in need thereof.

[0013] Also provided herein are engineered immune cells that express CD19-directed chimeric antigen receptors encoded by the polynucleotides disclosed herein. In some embodiments, the engineered immune cells are natural killer (NK) cells. In some embodiments, the engineered cells are T cells, although in some embodiments, combinations of NK cells and T cells (and optionally other types of immune cells) are used. In some embodiments, the immune cells are allogeneic to the subject receiving the cells. Also provided herein is the use of immune cells that express CD19-directed chimeric antigen receptors encoded by the polynucleotides disclosed herein for the treatment of cancer in a mammal in need thereof. This document also provides the use of immune cells that express CD19-directed chimeric antigen receptors encoded by the polynucleotides disclosed herein for the manufacture of a medicament for the treatment of cancer in a mammal in need thereof.

[0014] In several embodiments, methods of treating cancer are provided using the polynucleotides disclosed herein. For example, in several embodiments, the methods comprise administering to a subject having a cancer a composition comprising a population of immune cells expressing CD19-directed chimeric antigen receptors as disclosed herein. In several embodiments, the CAR comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) of a single chain variable fragment (scFv) and a light chain variable domain (VL) of an scFv, a hinge, such as a CD8alpha hinge, a transmembrane domain, such as a CD8alpha transmembrane domain; and an intracellular signaling domain comprising an OX40 subdomain and a CD3zeta subdomain, and wherein the cell also expresses membrane-bound interleukin-15 (mbIL15). In some embodiments, the OX40 subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO: 5, the CD3zeta subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO: 7, and/or mbIL15 is encoded by a sequence having at least 95% sequence identity to SEQ ID NO: 11. In some embodiments, the encoded VH domain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 120, wherein the encoded VL domain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 118. In some embodiments, the polynucleotide encodes a CD19-directed chimeric antigen receptor having at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 187. As discussed above, in several embodiments, the immune cells expressing such anti-CD19 CAR constructs are natural killer (NK) cells. In some embodiments, the immune cells are T cells. In several embodiments, combinations of NK and T cells (and optionally other immune cells) are used. In several embodiments, the cells are allogeneic cells derived from a donor that is not the subject. In several embodiments, the cells are autologous cells derived from the subject. In some embodiments, mixtures of allogeneic and autologous cells can also be used. In several embodiments, the population administered comprises about 2 x 10 ⁶ cells per kilogram of body weight of the subject. In several embodiments, the administration is intravenous administration. In several embodiments, the method further comprises administering to the subject one or more doses of interleukin 2. In several embodiments, the methods also comprise administering to the subject another therapy. For example, in several embodiments, the methods comprise administering to the subject a chemotherapeutic treatment prior to the administration of the cells. In some embodiments, the chemotherapeutic treatment causes lymphodepletion in the subject. In some embodiments, the subject is administered a combination of cyclophosphamide and fludarabine prior to administration of the cells. In some embodiments, cyclophosphamide is administered at a dose of about 400 to about 600 mg/ ^m2 . In some embodiments, fludarabine is administered at a dose of about 25 to 25 mg/ ^m2 . In some embodiments, the chemotherapeutic lymphodepletion is performed several days prior to administration of the engineered immune cells, and optionally, is performed multiple times. For example, in some embodiments, the chemotherapeutic lymphodepletion is performed at least the fifth, fourth, and/or third day prior to administration of the engineered immune cells disclosed herein.

[0015] In some embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises a humanized CD19 binding moiety, a costimulatory domain, and a signaling domain. In some embodiments, the costimulatory domain comprises OX40. In some embodiments, the humanized CD19 binding moiety comprises a humanized scFv, wherein one or more of the heavy and light chains are humanized. In some embodiments, one or more of the CDRs of the heavy and/or light chains are humanized. For example, in several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH domain comprises a VH domain selected from SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123, and the VL domain comprises a VL domain selected from SEQ ID NO: 117, SEQ ID NO: 118, and SEQ ID NO: 119, a hinge and/or transmembrane domain, an intracellular signaling domain.

[0016] In several embodiments, a polynucleotide encoding a humanized chimeric antigen receptor (CAR) is provided, wherein the CAR comprises a single chain antibody or single chain antibody fragment that comprises a humanized CD19 binding domain, a transmembrane domain, a primary intracellular signaling domain comprising a native intracellular signaling domain of CD3-zeta or a functional fragment thereof, and a costimulatory domain comprising a native intracellular signaling domain of a protein selected from the group consisting of OX40, CD27, CD28, ICOS, and 4-1BB, or a functional fragment thereof, wherein said CD19 binding domain comprises a light chain complementarity determining region 1 (CDR1 LC) of SEQ ID NO: 124, 127, or 130, a light chain complementarity determining region 2 (CDR2 LC) of SEQ ID NO: 125, 128 or 131, and a light chain complementarity determining region 3 (CDR3 LC) of SEQ ID NO: 126, 129 or 132, and a heavy chain complementarity determining region 1 (CDR1 HC) of SEQ ID NO: 133, 136, 139 or 142, a heavy chain complementarity determining region 2 (CDR2 HC) of SEQ ID NO: 134, 137, 140 or 143 and a heavy chain complementarity determining region 3 (CDR3 HC) of SEQ ID NO: 135, 138, 141 or 144.

[0017] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a light chain variable domain (VL) of SEQ ID NO: 117, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 161, SEQ ID NO: 167, SEQ ID NO: 173, SEQ ID NO: 179, SEQ ID NO: 185, SEQ ID NO: 191, SEQ ID NO: 197, or SEQ ID NO: 203.

[0018] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a light chain variable domain (VL) of SEQ ID NO: 118, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 163, SEQ ID NO: 169, SEQ ID NO: 175, SEQ ID NO: 181, SEQ ID NO: 187, SEQ ID NO: 193, SEQ ID NO: 199, or SEQ ID NO: 205.

[0019] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a light chain variable domain (VL) of SEQ ID NO: 119, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 165, SEQ ID NO: 171, SEQ ID NO: 177, SEQ ID NO: 183, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 201, or SEQ ID NO: 207.

[0020] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 120, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 185, SEQ ID NO: 187, or SEQ ID NO: 189.

[0021] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 121, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171, SEQ ID NO: 191, SEQ ID NO: 193, or SEQ ID NO: 195.

[0022] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 122, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 173, SEQ ID NO: 175, SEQ ID NO: 177, SEQ ID NO: 197, SEQ ID NO: 199, or SEQ ID NO: 201.

[0023] In several embodiments, a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 123, a hinge and/or transmembrane domain, and an intracellular signaling domain. In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 179, SEQ ID NO: 181, SEQ ID NO: 183, SEQ ID NO: 203, SEQ ID NO: 205, or SEQ ID NO: 207.

[0024] In several embodiments, the provided polynucleotides also encode membrane-associated interleukin-15 (mbIL15).

[0025] In some embodiments, the intracellular signaling domain comprises an OX40 subdomain. However, in some embodiments, the intracellular signaling domain comprises one or more of an OX40 subdomain, a CD28 subdomain, an iCOS subdomain, a CD28-41BB subdomain, a CD27 subdomain, a CD44 subdomain, or combinations thereof.

[0026] In several embodiments, the chimeric antigen receptor comprises a hinge and a transmembrane domain, wherein the hinge is a CD8-alpha hinge, wherein the transmembrane domain is either a CD8-alpha transmembrane domain or an NKG2D transmembrane domain. In several embodiments, the intracellular signaling domain comprises a CD3-zeta domain.

[0027] In several embodiments, the polynucleotide does not encode a sequence of SEQ ID NO: 112, 113, or 114. In several embodiments, the polynucleotide does not encode a sequence of SEQ ID NO: 116.

[0028] In several embodiments, engineered NK cells, engineered T cells, and/or mixed populations of NK cells and T cells are provided that express one or more of the humanized CD19-directed chimeric antigen receptors provided herein.

[0029] Also provided are methods of treating cancer in a subject comprising administering to a subject having cancer engineered NK and/or T cells expressing the chimeric antigen receptors disclosed herein. Also provided are the use of the polynucleotides provided herein for treating cancer, as well as the use of the polynucleotides provided herein in the manufacture of a medicament for treating cancer.

[0030] Also provided herein in several embodiments is a polynucleotide encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises an scFv, a transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises a CD28 costimulatory domain and a CD3-zeta signaling domain.

[0031] Also provided herein in several embodiments is a polynucleotide encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises an scFv, a hinge, wherein the hinge is a CD8-alpha hinge, a transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises a CD3-zeta ITAM.

[0032] Also provided herein in several embodiments is a polynucleotide encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable region from an scFv or a light chain variable region from an scFv, a hinge, wherein the hinge is a CD8-alpha hinge, a transmembrane domain, wherein the transmembrane domain comprises a CD8-alpha transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises a CD3-zeta ITAM.

[0033] In some embodiments, the transmembrane domain comprises a CD8 alpha transmembrane domain. In some embodiments, the transmembrane domain comprises an NKG2D transmembrane domain. In some embodiments, the transmembrane domain comprises a CD28 transmembrane domain.

[0034] In several embodiments, the intracellular signaling domain comprises or further comprises a CD28 signaling domain. In several embodiments, the intracellular signaling domain comprises or further comprises a 4-1BB signaling domain. In several embodiments, the intracellular signaling domain comprises or further comprises an OX40 domain. In several embodiments, the intracellular signaling domain comprises or further comprises a 4-1BB signaling domain. In several embodiments, the intracellular signaling domain comprises or further comprises a domain selected from ICOS, CD70, CD161, CD40L, CD44, and combinations thereof.

[0035] In some embodiments, the polynucleotide further encodes a truncated epidermal growth factor receptor (EGFRt). In some embodiments, EGFRt is expressed in a soluble factor in the cell. In some embodiments, EGFRt is expressed in a membrane-bound form. In some embodiments, EGFRt functions to provide a "suicide switch" in the engineered NK cells. In some embodiments, the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). Also provided herein are engineered immune cells (e.g., NK or T cells, or mixtures thereof) that express a CD19-directed chimeric antigen receptor encoded by a polynucleotide disclosed herein. Further provided are methods of treating cancer in a subject comprising administering to a subject having a cancer engineered immune cells expressing the chimeric antigen receptors disclosed herein. Several embodiments provide for the use of the polynucleotides disclosed herein in the treatment of cancer and/or in the manufacture of a medicament for the treatment of cancer.

[0036] In some embodiments, the CD19 binding fragment comprises a heavy chain variable domain (VH) and a light chain variable domain (VL). In some embodiments, the VH domain has at least 95% (e.g., 95, 96, 97, 98, or 99%) identity to the VH domain amino acid sequence set forth in SEQ ID NO: 33. In some embodiments, the VL domain has at least 95% (e.g., 95, 96, 97, 98, or 99%) identity to the VL domain amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the CD19 binding moiety is derived from the VH and/or VL sequences of SEQ ID NO: 33 or 32. For example, in some embodiments, the VH and VL sequences of SEQ ID NO: 33 and/or 32 have been humanized and are therefore more readily expressed and/or are less immunogenic when administered to human subjects. Thus, in several embodiments, the CD19 binding moiety does not comprise SEQ ID NO: 32 and/or SEQ ID NO: 33. In several embodiments, the CD19 binding moiety comprises an scFv that targets CD19, wherein the scFv comprises a heavy chain variable region comprising the sequence of SEQ ID NO: 35 or a sequence that is at least 95% (e.g., 95, 96, 97, 98, or 99%) identical to SEQ ID NO: 35. In several embodiments, the CD19 binding moiety comprises an scFv that targets CD19, comprises a light chain variable region comprising the sequence of SEQ ID NO: 36 or a sequence that is at least 95% (e.g., 95, 96, 97, 98, or 99%) identical to SEQ ID NO: 36. In several embodiments, the CD19 binding moiety comprises a light chain CDR comprising a first, second and third complementarity determining region (respectively CDR1 LC, CDR2 LC and CDR3 LC) and/or a heavy chain CDR comprising a first, second and third complementarity determining region (respectively CDR1 HC, CDR2 HC and CDR3 HC). Depending on the embodiment, various combinations of CDR LC and CDR HC are used. For example, in one embodiment, the CD19 binding moiety comprises CDR1 LC, CDR3 LC, CD2 HC and CDR3 HC. In some embodiments, other combinations are used. In several embodiments, CDR1 LC comprises a sequence of SEQ ID NO: 37 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 37. In several embodiments, CDR2 LC comprises a sequence of SEQ ID NO: 38 or a sequence that is at least about 95% (e.g., 96, 97, 98, or 99%) homologous to the sequence of SEQ NO: 38. In several embodiments, CDR3 LC comprises a sequence of SEQ ID NO: 39 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 39. In several embodiments, CDR1 HC comprises a sequence of SEQ ID NO: 40 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 40. In several embodiments, CDR2 HC comprises a sequence of SEQ ID NO: 41, 42, or 43 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 41, 42, or 43. In several embodiments, HC CDR3 comprises a sequence of SEQ ID NO: 44 or a sequence that is at least about 95% (e.g., 96, 97, 98, 99, or 99%) homologous to the sequence of SEQ NO: 44.

[0037] In several embodiments, a CD19 binding fragment is also provided that comprises a light chain variable region (VL) and a heavy chain variable region (HL), wherein the VL region comprises first, second, and third complementarity determining regions (VL CDR1, VL CDR2, and VL CDR3, respectively), and the VH region comprises first, second, and third complementarity determining regions (VH CDR1, VH CDR2, and VH CDR3, respectively). In some embodiments, the VL region comprises a sequence of SEQ ID NO: 45, 46, 47, or 48, or a sequence that is at least about 95% (e.g., 96, 97, 98, 99, or 99%) homologous to a sequence of SEQ NO: 45, 46, 47, or 48. In some embodiments, the VH region comprises a sequence of SEQ ID NO: 49, 50, 51, or 52, or a sequence that is at least about 95% (e.g., 96, 97, 98, 99, or 99%) homologous to a sequence of SEQ NO: 49, 50, 51, or 52.

[0038] In some embodiments, a CD19 binding moiety is also provided that comprises a light chain CDR comprising a first, second, and third complementarity determining region (CDR1 LC, CDR2 LC, and CDR3 LC, respectively). In some embodiments, the CD19 binding moiety further comprises a heavy chain CDR comprising a first, second, and third complementarity determining region (CDR1 HC, CDR2 HC, and CDR3 HC, respectively). In several embodiments, CDR1 LC comprises a sequence of SEQ ID NO: 53 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 53. In several embodiments, CDR2 LC comprises a sequence of SEQ ID NO: 54 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 54. In several embodiments, CDR3 LC comprises a sequence of SEQ ID NO: 55 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 55. In several embodiments, CDR1 HC comprises a sequence of SEQ ID NO: 56 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 56. In several embodiments, CDR2 HC comprises a sequence of SEQ ID NO: 57 or a sequence that is at least about 95% homologous to the sequence of SEQ NO: 57. In several embodiments, the HC CDR3 comprises a sequence of SEQ ID NO: 58 or a sequence that is at least about 95% homologous to the sequence of SEQ ID NO: 58. In several embodiments, the CD19 binding moiety (and thus the resulting CAR) is engineered to not include certain sequences, such as, for example, those that may confer an increased risk of immunogenicity and/or side effects such as cytokine release syndrome. Thus, in several embodiments, the CD19 binding moiety does not comprise one or more of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 32 or SEQ ID NO: 33.

[0039] In some embodiments, the intracellular signaling domain of the chimeric receptor comprises an OX40 subdomain. In some embodiments, the intracellular signaling domain further comprises a CD3-zeta subdomain. In some embodiments, the OX40 subdomain comprises the amino acid sequence of SEQ ID NO: 16 (or a sequence that is at least about 95% homologous to the sequence of SEQ ID NO: 16), and the CD3-zeta subdomain comprises the amino acid sequence of SEQ ID NO: 8 (or a sequence that is at least about 95% homologous to the sequence of SEQ ID NO: 8).

[0040] In several embodiments, the hinge domain comprises a CD8a hinge domain. In several embodiments, the CD8a hinge domain comprises the amino acid sequence of SEQ ID NO: 2 or a sequence that is at least about 95% homologous to the sequence of SEQ ID NO: 2).

[0041] In several embodiments, the immune cell also expresses membrane-bound interleukin-15 (mbIL15). In several embodiments, mbIL15 comprises the amino acid sequence of SEQ ID NO: 12 or a sequence that is at least about 95% homologous to the sequence of SEQ ID NO: 12.

[0042] In some embodiments, the chimeric receptor further comprises an extracellular domain of an NKG2D receptor. In some embodiments, the immune cell expresses a second chimeric receptor comprising an extracellular domain of an NKG2D receptor, a transmembrane domain, a cytotoxic signaling complex, and optionally mbIL15. In some embodiments, the extracellular domain of the NKG2D receptor comprises a functional fragment of NKG2D comprising the amino acid sequence of SEQ ID NO: 26 or a sequence that is at least about 95% homologous to the sequence of SEQ ID NO: 26. In various embodiments, the immune cell engineered to express the chimeric antigen receptor and/or chimeric receptors disclosed herein is an NK cell. In some embodiments, T cells are used. In some embodiments, combinations of NK and T cells (and/or other immune cells) are used.

[0043] In several embodiments, provided herein are methods of treating a cancer in a subject comprising administering to the subject having the cancer engineered immune cells that target CD19 as disclosed herein. Also provided herein is the use of the immune cells that target CD19 as disclosed herein for treating the cancer. Similarly, also provided herein is the use of the immune cells that target CD19 as disclosed herein in the manufacture of a medicament for treating the cancer. In several embodiments, the cancer being treated is acute lymphocytic leukemia.

[0044] Some embodiments of the methods and compositions described herein relate to an immune cell. In some embodiments, the immune cell expresses a CD19-directed chimeric receptor comprising an extracellular fragment of CD19, a hinge and/or transmembrane domain, and/or an intracellular signaling domain. In some embodiments, the immune cell is a natural killer (NK) cell. In some embodiments, the immune cell is a T cell.

[0045] In some embodiments, the hinge domain comprises a CD8a hinge domain. In some embodiments, the hinge domain comprises an Ig4 SH domain.

[0046] In some embodiments, the transmembrane domain comprises a CD8a transmembrane domain. In some embodiments, the transmembrane domain comprises a CD28 transmembrane domain. In some embodiments, the transmembrane domain comprises a CD3 transmembrane domain.

[0047] In some embodiments, the signaling domain comprises an OX40 signaling domain. In some embodiments, the signaling domain comprises a 4-1BB signaling domain. In some embodiments, the signaling domain comprises a CD28 signaling domain. In some embodiments, the signaling domain comprises an NKp80 signaling domain. In some embodiments, the signaling domain comprises a CD16 IC signaling domain. In some embodiments, the signaling domain comprises a CD3-zeta or CD3ζ ITAM signaling domain. In some embodiments, the signaling domain comprises a mbIL-15 signaling domain. In some embodiments, the signaling domain comprises a 2A cleavage domain. In some embodiments, the mIL-15 signaling domain is separated from the remainder or other portion of the CD19-directed chimeric receptor by a 2A cleavage domain.

[0048] Some embodiments relate to a method comprising administering an immune cell described herein to a subject in need thereof. In some embodiments, the subject has cancer. In some embodiments, the administration causes treatment, inhibition, or prevention of progression of the cancer.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

[0049] Figure 1A shows images of non-limiting examples of CD19-directed chimeric receptors.

[0050] Figure 1B provides images of additional non-limiting examples of CD19-directed chimeric receptors.

[0051] Figure 2 also provides images of non-limiting examples of CD19-directed chimeric receptors.

[0052] Figure 3A also provides images of non-limiting examples of CD19-directed chimeric receptors.

[0053] Figure 3B also provides images of non-limiting examples of CD19-directed chimeric receptors.

[0054] Figure 3C also provides images of non-limiting examples of CD19-targeted chimeric receptors.

[0055] Figure 3D also provides images of non-limiting examples of CD19-targeted chimeric receptors comprising humanized CD19 binding domains.

[0056] Figure 3E also provides images of non-limiting examples of CD19-targeted chimeric receptors comprising humanized CD19 binding domains.

[0057] Figure 3F also provides images of non-limiting examples of CD19-targeted chimeric receptors comprising humanized CD19 binding domains.

[0058] Figure 3G also provides images of non-limiting examples of CD19-targeted chimeric receptors comprising humanized CD19 binding domains without a marker sequence.

[0059] Figure 3H also provides images of non-limiting examples of CD19-targeted chimeric receptors comprising humanized CD19 binding domains without a marker sequence.

[0060] Figure 3I also provides images of non-limiting examples of CD19-targeted chimeric receptors comprising humanized CD19 binding domains without a marker sequence.

[0061] Figure 4 shows a schematic of a non-limiting NK cell expansion protocol.

[0062] Figure 5 shows a schematic of an experimental protocol for evaluating the efficacy of a CD19-directed chimeric antigen receptor in accordance with several embodiments disclosed herein.

[0063] Figure 6A shows in vivo data related to the antitumor effect of various non-restrictive CD19-directed chimeric receptors.

[0064] Figure 6B provides a summary of the antitumor effects of various non-restrictive CD19-targeted chimeric receptors.

[0065] Figure 7 shows data relating to the expression of various CD19-directed chimeric receptor constructs by NK cells.

[0066] Figure 8A shows raw fluorescence data (mean fluorescence intensity, MFI) related to the expression of selected CD19-targeted chimeric receptors.

[0067] Figure 8B depicts data relating to the expression of selected CD19-targeted chimeric receptors, displayed as the percentage of NK cells expressing the indicated receptor.

[0068] Figures 9A-9D provide data relating to cytotoxicity (against Nalm6 or Raji cells) for the designated CD19-directed chimeric receptors.

[0069] Figure 9E shows summary data relating to cytotoxicity (against Nalm6 cells) for the designated CD19-directed chimeric receptors at different effector:target ratios.

[0070] Figure 9F shows summary data relating to cytotoxicity (against Raji cells) for the designated CD19-directed chimeric receptors at different effector:target ratios.

[0071] Figure 10A shows data relating to the degree of increase in cytotoxicity (against Nalm6 cells) for the designated CD19-directed chimeric receptors 7 days after transduction.

[0072] Figure 10B shows data relating to the degree of increase in cytotoxicity (against Raji cells) for the designated CD19-directed chimeric receptors 7 days after transduction.

[0073] Figure 10C shows data relating to the degree of increase in cytotoxicity (against Nalm6 cells) for the indicated CD19-directed chimeric receptors 14 days after transduction.

[0074] Figure 10D shows data relating to the degree of increase in cytotoxicity (against Raji cells) for the designated CD19-targeted chimeric receptors 14 days after transduction.

[0075] Figures 11A-11E depict data relating to cytokine release by NK cells expressing various CD19-directed chimeric receptors when co-cultured with Nalm6 cells. Figure 11A depicts granzyme B release. Figure 11B depicts perforin release. Figure 11C depicts TNF-alpha release. Figure 11D depicts GM-CSF release. Figure 11E depicts interferon-gamma release.

[0076] Figures 12A-12E depict data relating to cytokine release by NK cells expressing various CD19-directed chimeric receptors when co-cultured with Raji cells. Figure 12A depicts granzyme B release. Figure 12B depicts perforin release. Figure 12C depicts TNF-alpha release. Figure 12D depicts GM-CSF release. Figure 12E depicts interferon-gamma release.

[0077] Figure 13 shows in vivo imaging data relating to tumor burden over time in mice treated with PBS, untransduced NK cells, or NK cells expressing the indicated CD19-targeted chimeric receptors, with the indicated number of engineered NK cells administered (M = million cells).

[0078] Figure 14A shows the raw data relating to the detectable fluorescent signal from the in vivo data shown in Figure 13, with the data monitored for 25 days after administration of Nalm6 cells.

[0079] Figure 14B shows the data shown in Figure 14A on a logarithmic scale.

[0080] Figure 14C shows data relating to CD3 expression by said cells expressing said constructs.

[0081] Figure 14D shows data relating to CD56 expression by designated cells expressing designated constructs.

[0082] Figure 14E shows data relating to tumor cells expressing GFP when contacted with said cells expressing said constructs.

[0083] Figure 14F shows data relating to CD19 expressing tumor cells when contacted with said cells expressing said constructs.

[0084] Figure 15 presents data relating to selected functional characteristics of the selected humanized CD19 CAR constructs.

[0085] Figure 16 provides data relating to the expression of various humanized anti-CD19 CAR constructs disclosed in embodiments herein.

[0086] Figures 17A-17E show cytotoxicity data. Figure 17A shows data relating to the cytotoxicity of NK cells from a first donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells. Figure 17A shows data relating to the cytotoxicity of NK cells from a first donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Raji cells. Figure 17C shows data relating to the cytotoxicity of NK cells from a second donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells. Figure 17D shows data relating to the cytotoxicity of NK cells from a third donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Raji cells. Figure 17E shows data relating to the cytotoxicity of NK cells from a third donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells.

[0087] Figure 18 shows summary (from 3 donors) expression data for NK cells expressing the indicated non-restrictive anti-CD19 CAR constructs 3 days post-transduction.

[0088] Figures 19A-19D show cytotoxicity data upon rechallenge. Figure 19A shows data pertaining to the cytotoxicity of NK cells from a first donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Raji cells when Raji cells were added to the NK cell culture on day 7 and again on day 14. Figure 19B shows data pertaining to the cytotoxicity of NK cells from a first donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells when Nalm6 cells were added to the NK cell culture on day 7 and again on day 14. Figure 19C shows data pertaining to the cytotoxicity of NK cells from a second donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Raji cells when Raji cells were added to the NK cell culture on day 7 and again on day 14. again on day 14. Figure 19D shows data relating to the cytotoxicity of NK cells from a second donor engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells when Nalm6 cells were added to the NK cell culture on day 7 and again on day 14.

[0089] Figures 20A-20B show expression data. Figure 20A shows fluorescence data from the expression of non-limiting examples of humanized anti-CD19 CAR constructs from two donors 10 days after transduction. Figure 20B shows data relating to the percentage of cells expressing CD19 (indicating the expression efficiency of a given anti-CD19 CAR.

[0090] Figures 21A-21B show cytotoxicity data. Figure 21A shows data pertaining to the cytotoxicity of NK cells (from the first of the two donors identified in Figure 20) engineered to express select humanized anti-CD19 CAR constructs disclosed herein against Raji cells when the Raji cells were added to the NK cell culture on day 7 and again on day 14. Figure 21B shows data pertaining to the cytotoxicity of NK cells (from the second of the two donors identified in Figure 20) engineered to express select humanized anti-CD19 CAR constructs disclosed herein against Raji cells when the Raji cells were added to the NK cell culture on day 7 and again on day 14.

[0091] Figures 22A-22E show data relating to cytokine release by NK cells expressing various CD19-directed chimeric receptors when co-cultured with Raji cells. Figure 22A shows the release of gamma interferon. Figure 22B shows the release of GM-CSF. Figure 22C shows the release of tumor necrosis factor. Figure 22D shows the release of perforin. Figure 22E shows the release of granzyme.

[0092] Figures 23A-23D show data relating to NK cell survival. Figure 23A shows a survival analysis of NK cells from a first donor engineered to express the disclosed non-limiting embodiments of the anti-CD19 CAR constructs at days 7, 13, 19, and 26 post-transduction. Figure 23B shows a survival analysis of NK cells from a second donor engineered to express the disclosed non-limiting embodiments of the anti-CD19 CAR constructs at days 7, 13, 19, and 26 post-transduction. Figure 23C shows a survival analysis of NK cells from a third donor engineered to express the disclosed non-limiting embodiments of the anti-CD19 CAR constructs at days 11, 19, and 26 post-transduction. Figure 23D shows a survival analysis of NK cells from a fourth donor engineered to express the indicated non-limiting embodiments of the anti-CD19 CAR constructs at days 11, 19, and 26 post-transduction. For each experiment, the medium was changed twice a week.

[0093] Figure 24 shows data relating to the percentage of NK cells expressing the designated non-limiting embodiments of the anti-CD19 CAR constructs 11 days after transduction.

[0094] Figures 25A-25I show data relating to the expression of CD19 by NK cells transduced with various non-limiting embodiments of the anti-CD19 CAR constructs disclosed herein. Figure 25A shows NK cells transduced with GFP as a control. Figure 25B shows the expression of CD19 in NK cells transduced with NK19-1. Figure 25C shows the expression of CD19 in NK cells transduced with NK19H-1. Figure 25D shows the expression of CD19 in NK cells transduced with NK19H-2. Figure 25E shows the expression of CD19 in NK cells transduced with NK19H-3. Figure 25F shows the expression of CD19 in NK cells transduced with NK19H-4. Figure 25G shows the expression of CD19 in NK cells transduced with NK19H-5. Figure 25H shows the expression of CD19 in NK cells transduced with NK19H-11. Figure 25I shows the expression of CD19 in NK cells transduced with NK19H-12.

[0095] Figures 26A-26D show cytotoxicity data. Figure 26A shows data pertaining to the cytotoxicity of NK cells (from a first donor) engineered to express select humanized anti-CD19 CAR constructs disclosed herein against Raji cells when Raji cells were added to the NK cell culture on day 7 and again on day 14. Figure 26B shows data pertaining to the cytotoxicity of NK cells (from a first donor) engineered to express select humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells when Nalm6 cells were added to the NK cell culture on day 7 and again on day 14. Figure 26C shows data pertaining to the cytotoxicity of NK cells (from a second donor) engineered to express select humanized anti-CD19 CAR constructs disclosed herein against Raji cells when Raji cells were added to the NK cell culture on day 7 and again on day 14. Figure 26D shows data relating to the cytotoxicity of NK cells (from a second donor) engineered to express selected humanized anti-CD19 CAR constructs disclosed herein against Nalm6 cells when Nalm6 cells were added to the NK cell culture on day 7 and again on day 14.

[0096] Figures 27A-27B show data relating to the expression of CD19 by engineered NK cells. Figure 27A shows flow cytometry data for NK cells from a donor that were engineered to express various non-restrictive anti-CD19 CAR constructs disclosed herein. Figure 27B shows corresponding data for NK cells isolated from an additional donor.

[0097] Figures 28A-28B show cytotoxicity data. Figure 28A shows the number of Raji cells over time when exposed (at an E:T ratio of 1:1) to NK cells (from the first donor indicated in Figure 27) expressing non-limiting examples of the disclosed anti-CD19 CAR constructs. Figure 28B shows the number of Raji cells over time when exposed (at an E:T ratio of 1:1) to NK cells (from the second donor indicated in Figure 27) expressing non-limiting examples of the disclosed anti-CD19 CAR constructs.

[0098] Figures 29A-29E show data related to various humanized CD19-targeted CAR constructs and their efficacy in an in vivo model. Figure 29A shows a schematic representation of the experimental protocol for evaluating the efficacy of the CD19-targeted CAR constructs in vivo . Figure 29B shows an in vivo bioluminescence image of mice injected with NALM6 tumor cells and treated with the construct. Figure 29C shows a line graph of the bioluminescence data collected based on Figure 29B. Figure 29D shows a survival curve showing the days that animals in each treatment group survived. Figure 29E shows the relative expression level of the constructs by NK cells as measured by the MFI of the label included in the CD19 CAR construct.

[0099] Figures 30A-30F depict data relating to the expression of the disclosed CD19-directed CAR constructs. Figure 30A depicts data relating to the expression of the disclosed constructs as a percentage of all CD56-positive cells in a blood sample from mice 15 days after NK cell administration (e.g., according to the scheme depicted in Figure 29A). Figure 30B depicts data relating to the expression of the disclosed constructs as a percentage of cells that are both CD56-positive and express the Flag tag (as part of the anti-CD19 CAR construct), also 15 days after NK cell administration. Figure 30C depicts data relating to the detection of GFP+ tumor cells in samples from animals treated with the disclosed constructs, also 15 days after NK cell administration. Figures 30D, 30E and 30F show the corresponding data 32 days after NK cell administration.

[00100] Figures 31A-31B show data relating to the expression of untagged anti-CD19 CAR constructs. As discussed herein, in several embodiments, the anti-CD19 CAR constructs comprise a Flag or other tag for detection purposes. However, all constructs disclosed herein as comprising a tag are also included herein as being untagged. Figure 31A shows data relating to the expression of selected humanized anti-CD19 CAR constructs without a flag tag over time. Figure 31B shows corresponding data regarding the detected mean fluorescence intensity for the constructs.

[00101] Figures 32A-32D show data from an in vitro rechallenge experiment in which a first population of tumor cells was co-cultured with NK cells expressing the constructs and the NK cells were "rechallenged" with an additional bolus dose of tumor cells. Figure 32A shows data obtained 10 days after the start of co-culture with Raji cells. Figure 32B shows data for Raji cells at the final time point, 14 days. Figure 32C shows data for Nalm6 cells at 10 days. Figure 32D shows data for Nalm6 cells at 14 days.

[00102] Figures 33A-33J depict data pertaining to the assessment of cytokine production by NK cells expressing the constructs. Figure 33A shows the expression of gamma interferon by NK cells expressing the constructs following co-culture with Raji cells. Figure 33B shows the expression of GM-CSF by NK cells expressing the constructs following co-culture with Raji cells. Figure 33C shows the expression of tumor necrosis factor alpha by NK cells expressing the constructs following co-culture with Raji cells. Figure 33D shows the expression of perforin by NK cells expressing the constructs following co-culture with Raji cells. Figure 33E shows granzyme B expression by NK cells expressing the indicated constructs following co-culture with Raji cells. Figure 33F shows interferon-gamma expression by NK cells expressing the indicated constructs following co-culture with Nalm6 cells. Figure 33G shows GM-CSF expression by NK cells expressing the indicated constructs following co-culture with Nalm6 cells. Figure 33H shows tumor necrosis factor-alpha expression by NK cells expressing the indicated constructs following co-culture with Nalm6 cells. Figure 33I shows perforin expression by NK cells expressing the indicated constructs following co-culture with Nalm6 cells. Figure 33J shows granzyme B expression by NK cells expressing the indicated constructs after co-culture with Nalm6 cells.

[00103] Figure 34 shows data relating to the viability of NK cells expressing humanized untagged anti-CD19 CAR constructs for four weeks after transduction.

[00104] Figures 35A-35D show data related to various humanized unlabeled CD19-targeted CAR constructs and their efficacy in an in vivo model. Figure 35A shows a schematic representation of the experimental protocol for evaluating the efficacy of the humanized unlabeled CD19-targeted CAR constructs in vivo . Figure 35B shows an in vivo bioluminescence image of mice injected with NALM6 tumor cells and treated with the construct. Figure 35C shows a line graph of the bioluminescence data collected from Figure 35B. Figure 35D shows the relative expression level of the constructs by NK cells as measured by detection of the CD19-Fc fusion protein that binds to the anti-CD19 CAR construct.

[00105] Figures 36A-36F depict data relating to the expression of the disclosed CD19-directed CAR constructs. Figure 36A depicts data relating to the expression of the disclosed constructs as a percentage of all CD56 positive cells in a blood sample from mice 13 days after NK cell administration (e.g., according to the scheme depicted in Figure 35A). Figure 36B depicts data relating to the detection of CD19+ tumor cells in samples from animals treated with the disclosed constructs, also 13 days after NK cell administration. Figure 36C depicts data relating to the detection of GFP+ tumor cells in samples from animals treated with the disclosed constructs, also 13 days after NK cell administration. Figures 36D, 36E, and 36F depict corresponding data 27 days after NK cell administration.

[00106] Figures 37A-37C provide data relating to the in vivo efficacy of various CD19-targeted CARs according to embodiments disclosed herein. Figure 37A shows a schematic diagram of an experimental protocol for evaluating the in vivo efficacy of humanized NK cells expressing various CD19-targeted CAR constructs. The various experimental groups tested are indicated. For cells designated "IL12/IL18," the cells were expanded in the presence of soluble IL12 and/or IL18 as described in U.S. Provisional Patent Application No. 62/881,311, filed July 31, 2019, and U.S. Patent Application No. 62/932,342, filed November 7, 2019, each of which is incorporated herein by reference in its entirety. Figures 37B and 37C show bioluminescence data from animals dosed with Nalm6 tumor cells and treated with the construct.

[00107] Figures 38A-38J are graphical representations of the bioluminescence data based on Figure 37B. Figure 38A shows the bioluminescence (as photon flux per second) from animals receiving untransduced NK cells. Figure 38B shows the flux measured in animals receiving PBS as the vehicle. Figure 38C shows the flux measured in animals receiving pre-frozen NK cells expressing the NK19 NF2 CAR (as a non-limiting example of a CAR). Figure 38D shows the flux measured in animals receiving pre-frozen NK cells expressing the NK19 NF2 CAR (as a non-limiting example of a CAR) expanded with IL12 and/or IL18. Figure 38E and Figure 38F show the flux measured in animals receiving fresh NK cells expressing the NK19 NF2 CAR (as a non-limiting example of a CAR). Figure 38G and Figure 38H show the flux measured in animals previously receiving fresh NK cells expressing the NK19 NF2 CAR (as a non-limiting example of a CAR) expanded with IL12 and/or IL18. Figure 38I shows a line graph depicting the bioluminescence measured in different groups during the first 30 days after tumor inoculation. Figure 38J shows a line graph depicting the bioluminescence measured in different groups during the first 56 days after tumor inoculation.

[00108] Figure 39 shows data relating to body weight of mice over time while receiving the indicated therapy.

DETAILED DESCRIPTION

[00109] Some embodiments of the methods and compositions provided herein relate to CD19-directed chimeric receptors. In some embodiments, the receptors are expressed in a cell described herein. Some embodiments include methods of using the compositions or cells in immunotherapy.

[00110] The term "anti-cancer effect" refers to a biological effect that may be manifested in a variety of ways, including, but not limited to, reducing tumor volume, reducing the number of cancer cells, reducing the number of metastases, increasing life expectancy, reducing cancer cell proliferation, reducing cancer cell survival, or reducing the intensity of various physiological symptoms associated with the cancerous condition. An "anti-cancer effect" may also be manifested in the ability of SIR to prevent cancer from occurring in the first place.

Cell types

[00111] Some embodiments of the methods and compositions provided herein relate to a cell, such as an immune cell. For example, an immune cell can be engineered to include a chimeric receptor, such as a CD19-directed chimeric receptor, or can be engineered to include a nucleic acid encoding the chimeric receptor described herein.

[00112] Traditional cancer therapies rely on surgery, radiation therapy, chemotherapy, or a combination of these methods. As research has led to a better understanding of some of the mechanisms of certain cancers, this knowledge has been used to develop targeted cancer therapies. Targeted therapy is a cancer treatment that uses specific drugs to specifically target specific genes or proteins found in cancer cells or cells that support cancer growth (such as blood vessel cells) to reduce or stop the growth of cancer cells. More recently, genetic engineering has made it possible to develop approaches that use certain aspects of the immune system to combat cancers. In some cases, a patient's own immune cells are modified to specifically kill a type of cancer in that patient. Various types of immune cells can be used, such as T cells or natural killer (NK) cells, which are described in more detail below.

[00113] To provide cancer immunotherapies, provided herein are polynucleotides, polypeptides, and vectors that encode chimeric antigen receptors (CARs) that comprise a target binding moiety (e.g., an extracellular binding portion of a ligand or a CD19-directed chimeric receptor expressed under the influence of a cancer cell) and a cytotoxic signaling complex. For example, some embodiments include a polynucleotide, polypeptide, or vector that encodes a CD19-directed chimeric receptor to provide for the targeting of an immune cell to a cancer and the exertion of cytotoxic effects on a cancer cell. Also provided are engineered immune cells (e.g., T cells or NK cells) expressing such CARs. In several embodiments, provided herein are also polynucleotides, polypeptides, and vectors that encode a construct comprising an extracellular domain comprising two or more subdomains, such as a first subdomain targeting CD19 comprising a CD19 binding moiety disclosed herein and a second subdomain comprising a C-type lectin-like receptor and a cytotoxic signaling complex. Also provided are engineered immune cells (e.g., T cells or NK cells) expressing such bispecific constructs. Also provided are engineered immune cells (e.g., T cells or NK cells) expressing multispecific constructs and/or having the ability to bind multiple target markers. Also provided herein are methods of treating cancer and other uses of such cells for cancer immunotherapy.

Engineered cells for immunotherapy

[00114] In several embodiments, immune system cells are engineered to have enhanced cytotoxic effects on target cells, such as tumor cells. For example, an immune system cell can be engineered to contain a CD19-directed chimeric receptor as described herein. In several embodiments, white blood cells or leukocytes are used because their natural function is to protect the body from abnormal cell growth and infectious diseases. There are many types of white blood cells that serve specific functions in the human immune system and are therefore a preferred starting point for engineering the cells described herein. White blood cells include granulocytes and agranulocytes (based on the presence or absence of granules in the cytoplasm, respectively). Granulocytes include basophils, eosinophils, neutrophils, and mast cells. Agranulocytes include lymphocytes and monocytes. Cells such as those described below or otherwise herein can be engineered to contain a chimeric receptor, such as a CD19-directed chimeric receptor, or a nucleic acid encoding a chimeric receptor, and/or engineered to co-express the costimulatory domain of membrane-bound interleukin 15 (mbIL15).

Monocytes for immunotherapy

[00115] Monocytes are a subtype of leukocytes. Monocytes can differentiate into macrophages and dendritic cells of myeloid origin. Monocytes are associated with the adaptive immune system and perform the essential functions of phagocytosis, antigen presentation, and cytokine production. Phagocytosis is the process of engulfing cellular material or entire cells, followed by digestion and destruction of the engulfed cellular material. In some embodiments, monocytes are used in conjunction with one or more additional engineered cells disclosed herein. Some embodiments of the methods and compositions described herein relate to a monocyte that comprises a CD19-targeted chimeric receptor or a nucleic acid encoding a CD19-targeted chimeric receptor. Several embodiments of the methods and compositions disclosed herein relate to monocytes engineered to express a CD19-directed chimeric receptor and the costimulatory domain of membrane-bound interleukin 15 (mbIL15).

Lymphocytes for immunotherapy

[00116] Lymphocytes, another primary subtype of leukocytes, include T cells (cell-mediated, cytotoxic adaptive immunity), natural killer cells (cell-mediated, cytotoxic innate immunity), and B cells (humoral, antibody-regulated adaptive immunity). Although B cells are engineered according to several embodiments disclosed herein, several embodiments also relate to engineered T cells or engineered NK cells (in some embodiments, mixtures of T cells and NK cells are used). Some embodiments of the methods and compositions described herein relate to a lymphocyte that comprises a CD19-directed chimeric receptor or a nucleic acid encoding a CD19-directed chimeric receptor. Several embodiments of the methods and compositions disclosed herein relate to lymphocytes engineered to express a CD19-directed chimeric receptor and the costimulatory domain of membrane-bound interleukin 15 (mbIL15).

T cells for immunotherapy

[00117] T cells are distinguished from other lymphocyte subtypes (e.g., B cells or NK cells) by the presence of a T cell receptor on the cell surface. T cells can be divided into various subtypes, including effector T cells, helper T cells, cytotoxic T cells, memory T cells, regulatory T cells, natural killer T cells, mucosal-associated invariant T cells, and gamma/delta T cells. In some embodiments, a specific T cell subtype is engineered. In some embodiments, a mixed pool of T cell subtypes is engineered. In some embodiments, there is no specific selection of the type of T cell to be engineered to express the cytotoxic receptor complexes disclosed herein. In some embodiments, specific techniques such as cytokine stimulation are used to enhance the expansion/harvesting of T cells with a specific marker profile. For example, in some embodiments, activation of certain human T cells, such as CD4+ T cells, CD8+ T cells, is achieved by using CD3 and/or CD28 as stimulatory molecules. In some embodiments, a method of treating or preventing cancer or an infectious disease is provided, comprising administering a therapeutically effective amount of T cells expressing a cytotoxic receptor complex and/or a homing fragment as described herein. In some embodiments, the engineered T cells are autologous cells, while in some embodiments, the T cells are allogeneic cells. Some embodiments of the methods and compositions described herein relate to a T cell that comprises a CD19-directed chimeric receptor or a nucleic acid encoding a CD19-directed chimeric receptor. Several embodiments of the methods and compositions disclosed herein relate to T cells engineered to express a CD19-directed chimeric receptor and the costimulatory domain of membrane-bound interleukin 15 (mbIL15).

NK cells for immunotherapy

[00118] In several embodiments, a method of treating or preventing cancer or an infectious disease is provided, comprising administering a therapeutically effective amount of natural killer (NK) cells expressing a cytotoxic receptor complex and/or a homing fragment as described herein. In several embodiments, the engineered NK cells are autologous cells, while in some embodiments, the NK cells are allogeneic cells. In several embodiments, NK cells are preferred because the natural cytotoxic potential of NK cells is relatively high. In several embodiments, it is surprisingly useful that the engineered cells disclosed herein can further enhance the cytotoxic activity of NK cells, resulting in even more effective activity against target cells (e.g., tumor or other diseased cells). Some embodiments of the methods and compositions described herein relate to NK that comprises a CD19-directed chimeric receptor or a nucleic acid encoding a CD19-directed chimeric receptor. Several embodiments of the methods and compositions disclosed herein relate to NK cells engineered to express a CD19-directed chimeric receptor and a costimulatory domain of membrane-bound interleukin 15 (mbIL15).

Hematopoietic stem cells for cancer immunotherapy

[00119] In some embodiments, the immunotherapy methods disclosed herein utilize hematopoietic stem cells (HSCs). In some embodiments, the cells are engineered to express a homing fragment and/or a cytotoxic receptor complex. In some embodiments, the HSCs are harnessed to utilize their engraftment capacity for long-term production of blood cells, which can provide a sustained source of targeted anti-cancer effector cells, such as to combat cancer remissions. In some embodiments, such continued production helps to compensate for immunological tolerance or exhaustion of other cell types, such as that associated with the tumor microenvironment. In some embodiments, allogeneic HSCs are used, while in some embodiments, autologous HSCs are used. In some embodiments, the HSCs are used in combination with one or more additional types of engineered cells disclosed herein. Some embodiments of the methods and compositions described herein relate to a stem cell, such as a hematopoietic stem cell, that comprises a CD19-targeted chimeric receptor or a nucleic acid encoding a CD19-targeted chimeric receptor. Several embodiments of the methods and compositions disclosed herein relate to stem cells, such as hematopoietic stem cells, that are engineered to express a CD19-targeted chimeric receptor and a costimulatory domain of membrane-bound interleukin 15 (mbIL15).

Extracellular domains (tumor-binding domains)

[00120] Some embodiments of the compositions and methods described herein relate to a chimeric receptor, such as a CD19-directed chimeric receptor, that comprises an extracellular domain. In some embodiments, the extracellular domain comprises a tumor-binding domain (also referred to as an antigen-binding protein or antigen-binding domain), as described herein. In some embodiments, the antigen-binding domain is derived from or comprises a wild-type or non-wild-type antibody, antibody fragment, scFv, Fv, Fab, (Fab')2, single-domain antibody (SDAB), vH or vL domain, camelid VHH domain, or a non-immunoglobulin backbone such as DARPIN, affibody, affilin, adnectin, affitin, repebody protein, finomer, alpha-body, avimer, atrimer, centirin, pronectin, anticalin, Kunitz domain, Armadillo repeat protein, self-antigen, receptor, or ligand. In some embodiments, the tumor-binding domain comprises more than one antigen-binding domain. In embodiments, the antigen binding domain is operably linked directly or via an optional linker to the NH2-terminal region of a TCR domain (e.g., the constant chains of TCR alpha, TCR beta, TCR beta2, pre-TCR alpha, pre-TCR alpha-Del48, TCR gamma, or TCR delta).

Antigen binding proteins

[00121] In several embodiments, antigen-binding proteins are provided. As used herein, the term "antigen-binding protein" shall have its ordinary meaning and shall also refer to a protein comprising an antigen-binding moiety that binds an antigen, and, optionally, a backbone or framework portion that allows the antigen-binding moiety to adopt a conformation that facilitates binding of the antigen-binding protein to the antigen. In some embodiments, the antigen is a cancer antigen (e.g., CD19) or a fragment thereof. In some embodiments, the antigen-binding moiety comprises at least one CDR from an antibody that binds the antigen. In some embodiments, the antigen-binding moiety comprises all three CDRs from the heavy chain of the antibody that binds the antigen or from the light chain of the antibody that binds the antigen. In yet other embodiments, the antigen-binding moiety comprises all six CDRs from the antibody that binds the antigen (three from the heavy chain and three from the light chain). In some embodiments, the antigen-binding fragment comprises one, two, three, four, five, or six CDRs from an antibody that binds to the antigen, and in some embodiments, the CDRs can be any combination of heavy and/or light chain CDRs. In some embodiments, the antigen-binding fragment is an antibody fragment.

[00122] Non-limiting examples of antigen-binding proteins include antibodies, antibody fragments (e.g., an antigen-binding fragment of an antibody), antibody derivatives, and antibody analogs. Additional specific examples include, but are not limited to, a single-chain variable fragment (scFv), a nanobody (e.g., a VH domain of camelid antibodies consisting of a heavy chain; a VHH fragment), a Fab fragment, a Fab' fragment, an F(ab')2 fragment, an Fv fragment, an Fd fragment, and a complementarity determining region (CDR) fragment. These molecules can be obtained from any mammalian source, such as a human, mouse, rat, rabbit, or pig, a dog, or a camelid. Antibody fragments can compete for binding to a target antigen with an intact (e.g., native) antibody, and the fragments can be produced by modifying intact antibodies (e.g., by enzymatic or chemical cleavage) or synthesized de novo using recombinant DNA or peptide synthesis technologies. The antigen-binding protein can comprise, for example, an alternative protein backbone or an artificial backbone with grafted CDRs or CDR derivatives. Such backbones include, but are not limited to, antibody-derived backbones that contain mutations introduced, for example, to stabilize the three-dimensional structure of the antigen-binding protein, as well as fully synthetic backbones comprising, for example, a biocompatible polymer. In addition, peptide antibody mimetics ("PAMs") can be used, as well as antibody mimetic-based backbones using fibronectin components as a backbone.

[00123] In some embodiments, an antigen-binding protein comprises one or more antibody fragments included in a single polypeptide chain or in multiple polypeptide chains. For example, antigen-binding proteins can include, but are not limited to, a diabody; an intrabody; a domain antibody (one VL or VH domain or two or more VH domains connected by a peptide linker); a maxibody (2 scFv fused to an Fc region); a triabody; a tetrabody; a minibody (an scFv fused to a CH3 domain); a peptide-associated antibody (one or more peptides attached to an Fc region); a linear antibody (a pair of tandem Fd segments (VH-CH1-VH-CH1) that, together with complementary light chain polypeptides, form a pair of antigen-binding regions); small modular protein-based immunopharmaceutical and immunoglobulin-based fusion proteins (e.g. IgG-scFv, IgG-Fab, 2scFv-IgG, 4scFv-IgG, VH-IgG, IgG-VH and Fab-scFv-Fc).

[00124] In some embodiments, the antigen-binding protein has an immunoglobulin structure. As used herein, the term "immunoglobulin" shall have its ordinary meaning and shall also refer to a tetrameric molecule, wherein each tetramer comprises two identical pairs of polypeptide chains, wherein each pair has one "light" (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100-110 or more amino acids, primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region, primarily responsible for effector function.

[00125] In the light and heavy chains, the variable (V) and constant (C) regions are connected by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 10 additional amino acids. The variable regions of each light/heavy chain pair form an antibody binding site such that an intact immunoglobulin contains two binding sites.

[00126] Immunoglobulin chains exhibit a similar overall structure of relatively conserved framework regions (FRs) connected by three hypervariable regions, also called complementarity determining regions or CDRs. From N-terminus to C-terminus, both light and heavy chains contain FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 domains.

[00127] Human light chains are classified as kappa and lambda light chains. The term "light chain" of an antibody refers to the smaller of the two types of polypeptide chains that are present in antibody molecules in their naturally occurring conformations. Kappa (K) and lambda (λ) light chains refer to the two major antibody light chain isotypes. A light chain may include a polypeptide comprising, from the amino terminus to the carboxyl terminus, one immunoglobulin light chain variable region (VL) and one immunoglobulin light chain constant domain (CL).

[00128] Heavy chains are classified as mu (μ), delta (Δ), gamma (γ), alpha (α), and epsilon (ε), and the isotype of an antibody is defined as IgM, IgD, IgG, IgA, and IgE, respectively. The term "heavy chain" of an antibody refers to the larger of the two types of polypeptide chains that are present in antibody molecules in their naturally occurring conformations and that generally defines the class to which the antibody belongs. A heavy chain may include a polypeptide comprising, from the amino terminus to the carboxyl terminus, one immunoglobulin heavy chain variable region (VH), an immunoglobulin heavy chain constant domain 1 (CH1), an immunoglobulin hinge region, an immunoglobulin heavy chain constant domain 2 (CH2), an immunoglobulin heavy chain constant domain 3 (CH3), and optionally an immunoglobulin heavy chain constant domain 4 (CH4).

[00129] The IgG class is further divided into subclasses, namely IgG1, IgG2, IgG3, and IgG4. The IgA class is further divided into subclasses, namely IgA1 and IgA2. The IgM class includes subclasses, including, but not limited to, IgM1 and IgM2. The heavy chains in IgG, IgA, and IgD antibodies contain three domains (CH1, CH2, and CH3), while the heavy chains in IgM and IgE antibodies contain four domains (CH1, CH2, CH3, and CH4). The constant domains of an immunoglobulin heavy chain can be of any immunoglobulin isotype, including subtypes. The antibody chains are linked to each other by interpolypeptide disulfide bonds between the CL domain and the CH1 domain (e.g., between the light and heavy chains) and between the hinge regions of the antibody heavy chains.

[00130] In some embodiments, the antigen-binding protein is an antibody. The term "antibody" as used herein refers to a protein or polypeptide sequence derived from an immunoglobulin molecule that specifically binds to an antigen. Antibodies may be monoclonal or polyclonal, multi- or single-chain, or intact immunoglobulins, and may be from natural sources or from recombinant sources. Antibodies may be tetramers of immunoglobulin molecules. An antibody may be "humanized," "chimeric," or non-human. An antibody may include intact immunoglobulin of any isotype, and includes, for example, chimeric, humanized, human, and bispecific antibodies. An intact antibody typically comprises at least two full-length heavy chains and two full-length light chains. Antibody sequences may be exclusively from one species or may be "chimeric," meaning different portions of the antibody may be from two different species, as described further below. Unless otherwise specified, the term "antibody" also includes antibodies comprising two substantially full-length heavy chains and two substantially full-length light chains, so long as the antibodies retain the same or similar binding properties and/or function as an antibody comprising two full-length light chains and heavy chains. For example, antibodies comprising substitutions, insertions, or deletions of 1, 2, 3, 4, or 5 amino acid residues at the N-terminus and/or C-terminus of the heavy and/or light chains are included within the definition, so long as the antibodies retain the same or similar binding properties and/or function as an antibody comprising two full-length heavy chains and two full-length light chains. Examples of antibodies include monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, bispecific antibodies, and synthetic antibodies. In some embodiments, monoclonal and polyclonal antibodies are provided. As used herein, the term "polyclonal antibody" shall have its ordinary meaning and shall also refer to a population of antibodies that typically vary widely in composition and binding specificity. As used herein, the term "monoclonal antibody" ("mAb") shall have its ordinary meaning and shall also refer to one or more of a population of antibodies that have identical sequences. Monoclonal antibodies bind to an antigen at a specific epitope of the antigen.

[00131] In some embodiments, the antigen-binding protein is a fragment or antigen-binding fragment of an antibody. The term "antibody fragment" refers to at least one portion of an antibody that retains the ability to specifically interact (e.g., by binding, steric isolation, stabilization/destabilization, spatial distribution) with an epitope of an antigen. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, scFv antibody fragments, disulfide-linked Fv (sdFv), an Fd fragment consisting of VH and CHI domains, linear antibodies, single-domain antibodies such as sdAbs (either vL or vH), camelid vHH domains, multispecific antibodies formed from antibody fragments such as a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and an isolated CDR or other epitope-binding fragments of an antibody. The antigen-binding moiety can also be included in single-domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR, and bis-scFv (see, e.g., Hollinger and Hudson, Nature Biotechnology 23: 1126-1136, 2005). Antigen-binding moieties can also be grafted to polypeptide backbones such as fibronectin type III (Fn3) (see U.S. Patent No. 6,703,199, which describes fibronectin polypeptide minibodies). The antibody moiety can include a Fab, Fab', F(ab')2, and/or Fv moiety that contains at least one immunoglobulin CDR that is sufficient to provide specific antigen binding to a cancer antigen (e.g., CD19). Antibody fragments can be produced using recombinant DNA technologies or by enzymatic or chemical cleavage of intact antibodies.

[00132] In some embodiments, Fab fragments are provided. The Fab fragment is a monovalent fragment characterized by VL, VH, CL, and CH1 domains; the F(ab')2 fragment is a bivalent fragment having two Fab fragments linked by a disulfide bridge at the hinge region; the Fd fragment comprises VH and CH1 domains; the Fv fragment comprises the VL and VH domains of one arm of an antibody; and the dAb fragment comprises a VH domain, a VL domain, or an antigen-binding fragment of a VH or VL domain. In some embodiments, these antibody fragments can be included in single-domain antibodies, single-chain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR, and bis-scFv. In some embodiments, the antibodies comprise at least one CDR described herein.

[00133] In several embodiments, single-chain variable fragments are also provided herein. As used herein, the term "single-chain variable fragment" ("scFv") shall have its ordinary meaning and shall also refer to a fusion protein in which a VL region and a VH region are joined by a linker (e.g., a synthetic sequence of amino acid residues) to form a continuous protein chain, wherein the linker is of sufficient length to allow the protein chain to fold onto itself and form a monovalent antigen-binding site). For the avoidance of doubt, unless otherwise indicated, a "single-chain variable fragment" is not an antibody or antibody fragment as defined herein. Diabodies are bivalent antibodies comprising two polypeptide chains, wherein each polypeptide chain comprises VH and VL domains connected by a linker configured to reduce or prevent pairing between the two domains on one chain, thereby allowing each domain to pair with a complementary domain on another polypeptide chain. According to several embodiments, if the two polypeptide chains of a diabody are identical, the diabody resulting from their pairing will have two identical antigen-binding sites. Polypeptide chains comprising different sequences can be used to create a diabody with two different antigen-binding sites. Similarly, triabodies and tetrabodies are antibodies comprising three and four polypeptide chains, respectively, and forming three and four antigen-binding sites, respectively, which may be the same or different.

[00134] In several embodiments, the antigen-binding protein comprises one or more CDRs. As used herein, the term "CDR" will have its ordinary meaning and will also refer to the complementarity determining region (also referred to as "minimal recognition units" or "hypervariable region") in the variable sequences of an antibody. CDRs allow the antigen-binding protein to specifically bind to a particular antigen of interest. There are three CDRs of the variable region of the heavy chain (CDRH1, CDRH2, and CDRH3) and three CDRs of the variable region of the light chain (CDRL1, CDRL2, and CDRL3). The CDRs in each of the two chains are typically aligned within framework regions to form a structure that specifically binds to a specific epitope or domain on a target protein. Naturally occurring variable regions of the light and heavy chains are typically arranged in the following order from N-terminus to C-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. A numbering system has been developed to assign numbers to the amino acids that occupy positions in each of these domains. This numbering system is defined in Kabat Sequences of Proteins of Immunological Interest (1987 and 1991, NIH, Bethesda, MD) or Chothia & Lesk, 1987, J. Mol. Biol. 196:901–917; Chothia et al., 1989, Nature 342:878–883. This system can be used to identify the complementarity determining regions (CDRs) and framework regions (FRs) of a given antibody. Other systems for numbering amino acids in immunoglobulin chains include IMGT® (International Immunogenetics Information System; Lefranc et al, Dev. Comp. Immunol. 29:185-203; 2005) and AHo (Honegger and Pluckthun, J. Mol. Biol. 309(3):657-670; 2001). One or more CDRs can be included in the molecule either covalently or non-covalently to make it an antigen-binding protein. In some embodiments, the antigen-binding proteins provided herein comprise a heavy chain variable region selected from SEQ ID NO: 104 and SEQ ID NO: 106. In some embodiments, the antigen-binding proteins provided herein comprise a light chain variable region selected from SEQ ID NO: 105 and SEQ ID NO: 107.

[00135] In several embodiments, the antigen-binding protein is modified compared to its base sequence, such as to improve expression, function, or reduce a potential host immune response to the antigen-binding protein. In several embodiments, the antigen-binding protein comprises a light chain variable region selected from SEQ ID NO: 117, SEQ ID NO: 118, and SEQ ID NO: 119 and/or sequences having at least 90% identity and/or homology (e.g., 90-95%, 95%, 96%, 97%, 98%, 99%). In some embodiments, the light chain variable region differs in sequence from SEQ ID NO: 117, SEQ ID NO: 118, or SEQ ID NO: 119 by more than 5% (e.g., 5-7%, 5-10%, 10-20%, or more), wherein the ligand binding function (or other functionality) is similar, substantially similar, or identical to SEQ ID NO: 117, SEQ ID NO: 118, or SEQ ID NO: 119. In some embodiments, the antigen-binding protein comprises a heavy chain variable region selected from SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123 and/or sequences having at least 90% identity and/or homology (e.g., 90-95%, 95%, 96%, 97%, 98%, 99%). In several embodiments, the heavy chain variable region differs in sequence from SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, or SEQ ID NO: 123 by more than 5% (e.g., 5-7%, 5-10%, 10-20%, or more), while the ligand binding function (or other functionality) is similar, substantially similar, or identical to SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, or SEQ ID NO: 123. Depending on the embodiment, any combination of heavy and light chain regions can be used (e.g., when assembling an scFv). In several embodiments, the antigen-binding protein comprises one or more CDRs selected from SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, and SEQ ID NO: 144.

[00136] In further embodiments, the CDRs are selected from SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, and SEQ ID NO: 115 in any combination. In one embodiment, the CDRs are assembled to form a CAR targeting CD19 and comprising SEQ ID NO: 116.

[00137] In some embodiments, the antigen-binding protein comprises a heavy chain having the sequence of SEQ ID NO: 88. In some embodiments, the heavy chain is linked (e.g., as an scFv) to one of the light chains of SEQ ID NO: 89, SEQ ID NO: 90, and/or SEQ ID NO: 91. In some embodiments, the antigen-binding protein comprises one or more CDRs selected from SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, and SEQ ID NO: 100.

[00138] In some embodiments, the antigen-binding protein comprises a light chain region of the FMC63 antibody that has the sequence of SEQ ID NO: 150. In some embodiments, the antigen-binding protein comprises a light chain region of the FMC63 antibody that has the sequence of SEQ ID NO: 148. In some embodiments, linkers are used between the heavy and light chains, and in some embodiments, the linker comprises the sequence of SEQ ID NO: 149. In some embodiments, such heavy and light chains are used in combination with a CD28 costimulatory domain, such as the domain of SEQ ID NO: 153. Often, a spacer is used to separate the constituent parts of the CAR. For example, in some embodiments, a spacer comprising the sequence of SEQ ID NO: 151 is used. In some embodiments, a transmembrane domain having the sequence of SEQ ID NO: 152 is used. In some embodiments, a CAR comprises a nucleic acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 147.

[00139] In some embodiments, the antigen-binding proteins provided herein comprise one or more CDRs as part of a larger polypeptide chain. In some embodiments, the antigen-binding proteins covalently link one or more CDRs to another polypeptide chain. In some embodiments, the antigen-binding proteins non-covalently include one or more CDRs. In some embodiments, the antigen-binding proteins may comprise at least one of the CDRs described herein included in a biocompatible framework. In some embodiments, the biocompatible framework comprises a polypeptide or a portion thereof sufficient to form a conformationally stable structural support or scaffold or backbone capable of reproducing one or more amino acid sequences that bind an antigen (e.g., a CDR, a variable region, etc.) in a localized surface region. Such structures may be a naturally occurring polypeptide or polypeptide "fold" (structural motif) or may include one or more modifications, such as additions, deletions, and/or substitutions of amino acids, compared to a naturally occurring polypeptide or fold. Depending on the embodiment, the scaffolds may be derived from polypeptides of multiple different species (or from more than one species), such as human, non-human primate or other mammal, other vertebrate, invertebrate, plant, bacteria, or virus.

[00140] Depending on the embodiment, the biocompatible scaffolds are based on protein backbones or skeletons other than immunoglobulin domains. In some such embodiments, the scaffolds are based on fibronectin, ankyrin, lipocalin, neocarzinostein, cytochrome b, zinc finger CP1, PST1, coiled coil, LACI-D1, Z domain, and/or tendamistat domains.

[00141] In some embodiments, antigen-binding proteins with more than one binding site are also provided. In some embodiments, the binding sites are identical to each other, while in some embodiments, the binding sites are different from each other. For example, an antibody typically contains two identical binding sites, while a "bispecific" or "bifunctional" antibody contains two different binding sites. The two binding sites of a bispecific antigen-binding protein or antibody will bind to two different epitopes, which may be on the same or different target proteins. In some embodiments, this is particularly advantageous because the bispecific chimeric antigen receptor can confer on the engineered cell the ability to target multiple tumor markers. For example, CD19 and an additional tumor marker such as CD123, NKG2D or any other marker disclosed herein or recognized in the art as a tumor-specific antigen or tumor-associated antigen.

[00142] As used herein, the term "chimeric antibody" shall have its ordinary meaning and shall also refer to an antibody that comprises one or more regions from one antibody and one or more regions from one or more other antibodies. In some embodiments, one or more CDRs are derived from an antibody to a cancer antigen (e.g., CD19). In some embodiments, all of the CDRs are derived from an antibody to a cancer antigen (such as an antibody to CD19). In some embodiments, CDRs from more than one antibody to a cancer antigen are mixed and matched in a chimeric antibody. For example, a chimeric antibody may comprise CDR1 from a light chain of a first antibody to a cancer antigen, CDR2 and CDR3 from a light chain of a second antibody to a cancer antigen, and a CDR from a heavy chain of a third antibody to a cancer antigen. Additionally, the framework regions of the antigen-binding proteins disclosed herein can be derived from one of the same antibodies to a cancer antigen (e.g., CD19), from one or more different antibodies, such as a human antibody, or from a humanized antibody. In one example of a chimeric antibody, a portion of the heavy and/or light chain is identical to, homologous to, or derived from an antibody from a particular species, or belongs to a particular class or subclass of antibodies, while the remainder of the chain(s) is identical to, homologous to, or derived from an antibody or antibodies from another species, or belongs to a different class or subclass of antibodies. Also provided herein are fragments of such antibodies that exhibit the desired biological activity.

[00143] In some embodiments, an antigen-binding protein is provided that comprises a heavy chain variable domain having at least 90% identity to the amino acid sequence of a VH domain set forth in SEQ ID NO: 33. In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 95% identity to the amino acid sequence of a VH domain set forth in SEQ ID NO: 33. In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 96%, 97%, 98%, or 99% identity to the amino acid sequence of a VH domain set forth in SEQ ID NO: 33. In some embodiments, the heavy chain variable domain may have one or more additional mutations (e.g., for humanization) in the amino acid sequence of the VH domain set forth in SEQ ID NO: 33, but retains specific binding to a cancer antigen (e.g., CD19). In some embodiments, the heavy chain variable domain may comprise one or more additional mutations in the VH domain amino acid sequence shown as SEQ ID NO: 33, but is characterized by improved specific binding to a cancer antigen (e.g., CD19).

[00144] In some embodiments, the antigen-binding protein comprises a light chain variable domain having at least 90% identity to the VL domain amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antigen-binding protein comprises a light chain variable domain having at least 95% identity to the VL domain amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antigen-binding protein comprises a light chain variable domain having at least 96%, 97%, 98%, or 99% identity to the VL domain amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the light chain variable domain may have one or more additional mutations (e.g., for humanization) in the VL domain amino acid sequence set forth in SEQ ID NO: 32, but retains specific binding to a cancer antigen (e.g., CD19). In some embodiments, the light chain variable domain may comprise one or more additional mutations in the VL domain amino acid sequence shown as SEQ ID NO: 32, but is characterized by improved specific binding to a cancer antigen (e.g., CD19).

[00145] In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 90% identity to the amino acid sequence of the VH domain set forth in SEQ ID NO: 33 and a light chain variable domain having at least 90% identity to the amino acid sequence of the VL domain set forth in SEQ ID NO: 32. In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 95% identity to the amino acid sequence of the VH domain set forth in SEQ ID NO: 33 and a light chain variable domain having at least 95% identity to the amino acid sequence of the VL domain set forth in SEQ ID NO: 32. In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 96, 97, 98, or 99% identity to the amino acid sequence of the VH domain presented under SEQ ID NO: 33, and a light chain variable domain characterized by at least 96, 97, 98 or 99% identity to the amino acid sequence of the VL domain presented under SEQ ID NO: 32.

[00146] In some embodiments, the antigen-binding protein comprises a heavy chain variable domain characterized by the VH domain amino acid sequence set forth in SEQ ID NO: 33 and a light chain variable domain characterized by the VL domain amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the light chain variable domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of the light chain variable domain of SEQ ID NO: 32. In some embodiments, the light chain variable domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the heavy chain variable domain sequence according to SEQ ID NO: 33.

[00147] In some embodiments, the light chain variable domain comprises an amino acid sequence that is encoded by a nucleotide sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the polynucleotide sequence of SEQ ID NO: 32. In some embodiments, the light chain variable domain comprises an amino acid sequence that is encoded by a polynucleotide that hybridizes under moderately stringent conditions to the complementary sequence of a polynucleotide that encodes the light chain variable domain of SEQ ID NO: 32. In some embodiments, the light chain variable domain comprises an amino acid sequence that is encoded by a polynucleotide that hybridizes under stringent conditions to the complementary sequence of a polynucleotide that encodes the light chain variable domain of SEQ ID NO: 32. a polynucleotide that encodes a light chain variable domain according to the sequence of SEQ ID NO: 32.

[00148] In some embodiments, the heavy chain variable domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of the heavy chain variable domain of SEQ ID NO: 33. In some embodiments, the heavy chain variable domain comprises an amino acid sequence that is encoded by a polynucleotide that hybridizes under moderately stringent conditions to the complementary sequence of a polynucleotide that encodes the heavy chain variable domain of SEQ ID NO: 33. In some embodiments, the heavy chain variable domain comprises an amino acid sequence that is encoded by a polynucleotide that hybridizes under moderately stringent conditions to the complementary a polynucleotide sequence that encodes a heavy chain variable domain according to the sequence of SEQ ID NO: 33.

[00149] In several embodiments, additional CD19 binding constructs are provided. For example, in some embodiments, an scFv is provided that targets CD19, wherein the scFv comprises a heavy chain variable region comprising the sequence of SEQ ID NO: 35. In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 95% identity to the amino acid sequence of the HCV domain set forth in SEQ ID NO: 35. In some embodiments, the antigen-binding protein comprises a heavy chain variable domain having at least 96, 97, 98, or 99% identity to the amino acid sequence of the HCV domain set forth in SEQ ID NO: 35. In some embodiments, the heavy chain variable domain may have one or more additional mutations (e.g., for humanization) in the amino acid sequence of the HCV domain set forth in SEQ ID NO: 35, but retains specific binding to a cancer antigen (e.g., CD19). In some embodiments, the heavy chain variable domain may comprise one or more additional mutations in the amino acid sequence of the HCV domain set forth in SEQ ID NO: 35, but is characterized by improved specific binding to a cancer antigen (e.g., CD19).

[00150] Additionally, in some embodiments, an scFv that targets CD19 comprises a light chain variable region comprising the sequence of SEQ ID NO: 36. In some embodiments, the antigen-binding protein comprises a light chain variable domain having at least 95% identity to the amino acid sequence of the LCV domain set forth in SEQ ID NO: 36. In some embodiments, the antigen-binding protein comprises a light chain variable domain having at least 96, 97, 98, or 99% identity to the amino acid sequence of the LCV domain set forth in SEQ ID NO: 36. In some embodiments, the light chain variable domain may have one or more additional mutations (e.g., for humanization) in the amino acid sequence of the LCV domain set forth in SEQ ID NO: 36, but retains specific binding to a cancer antigen (e.g., CD19). In some embodiments, the light chain variable domain may comprise one or more additional mutations in the amino acid sequence of the LCV domain as set forth in SEQ ID NO: 36, but is characterized by improved specific binding to a cancer antigen (e.g., CD19).

[00151] In some embodiments, a CD19 binding moiety is also provided that comprises a light chain CDR comprising a first, second, and third complementarity determining region (CDR1 LC, CDR2 LC, and CDR3 LC, respectively). In some embodiments, the CD19 binding moiety further comprises a heavy chain CDR comprising a first, second, and third complementarity determining region (CDR1 HC, CDR2 HC, and CDR3 HC, respectively). In some embodiments, CDR1 LC comprises the sequence of SEQ ID NO: 37. In some embodiments, CDR1 LC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 37. In some embodiments, CDR2 LC comprises the sequence of SEQ ID NO: 38. In some embodiments, CDR2 LC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 38. In some embodiments, CDR3 LC comprises the sequence of SEQ ID NO: 39. In some embodiments, CDR3 LC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 39. In some embodiments, CDR1 of HC comprises the sequence of SEQ ID NO: 40. In several embodiments, CDR1 of HC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 40. In several embodiments, CDR2 of HC comprises the sequence of SEQ ID NO: 41, 42, or 43. In several embodiments, CDR2 of HC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ ID NO: 41, 42, or 43. In several embodiments, CDR3 of HC comprises the sequence of SEQ ID NO: 44. In several embodiments, CDR3 of HC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ ID NO: 44.

[00152] In several embodiments, a CD19 binding fragment is also provided that comprises a light chain variable region (VL) and a heavy chain variable region (HL), wherein the VL region comprises first, second, and third complementarity determining regions (VL CDR1, VL CDR2, and VL CDR3, respectively), and the VH region comprises first, second, and third complementarity determining regions (VH CDR1, VH CDR2, and VH CDR3, respectively). In some embodiments, the VL region comprises a sequence of SEQ ID NO: 45, 46, 47, or 48. In some embodiments, the VL region comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 45, 46, 47, or 48. In some embodiments, the VH region comprises a sequence of SEQ ID NO: 49, 50, 51, or 52. In some embodiments, the VH region comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 49, 50, 51, or 52.

[00153] In some embodiments, a CD19 binding moiety is also provided that comprises a light chain CDR comprising a first, second, and third complementarity determining region (CDR1 LC, CDR2 LC, and CDR3 LC, respectively). In some embodiments, the CD19 binding moiety further comprises a heavy chain CDR comprising a first, second, and third complementarity determining region (CDR1 HC, CDR2 HC, and CDR3 HC, respectively). In some embodiments, CDR1 LC comprises the sequence of SEQ ID NO: 53. In some embodiments, CDR1 LC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 53. In some embodiments, CDR2 LC comprises the sequence of SEQ ID NO: 54. In some embodiments, CDR2 LC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 54. In some embodiments, CDR3 LC comprises the sequence of SEQ ID NO: 55. In some embodiments, CDR3 LC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 55. In some embodiments, CDR1 of HC comprises the sequence of SEQ ID NO: 56. In several embodiments, CDR1 of HC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 56. In several embodiments, CDR2 of HC comprises the sequence of SEQ ID NO: 57. In several embodiments, CDR2 of HC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 57. In several embodiments, CDR3 of HC comprises the sequence of SEQ ID NO: 58. In several embodiments, CDR3 of HC comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, or about 98% homologous to the sequence of SEQ NO: 58.

[00154] Additional CD19 binding moieties are known in the art, such as those disclosed, for example, in U.S. Patent No. 8,399,645, U.S. Patent Publication No. 2018/0153977, U.S. Patent Publication No. 2014/0271635, U.S. Patent Publication No. 2018/0251514, and U.S. Patent Publication No. 2018/0312588, each of which is incorporated herein by reference in its entirety.

Domains of natural killer receptor groups that bind tumor ligands

[00155] In several embodiments, engineered immune cells, such as NK cells, are used for their ability to recognize and kill tumor cells. For example, an engineered NK cell may include a CD19-directed chimeric receptor or a nucleic acid encoding the chimeric receptor. NK cells express both inhibitory and activating receptors on the cell surface. Inhibitory receptors bind self-molecules expressed on the surface of healthy cells (thereby preventing immune responses against "self" cells), while activating receptors bind ligands expressed on abnormal cells, such as tumor cells. When the balance between activation of inhibitory and activating receptors shifts in favor of activating receptors, NK cells are activated and target cells (e.g., tumor cells) are lysed.

[00156] Natural killer receptor group 2 D (NKG2D) is an activating NK cell receptor that recognizes a variety of ligands expressed on cells. Surface expression of various NKG2D ligands is typically low in healthy cells, but is upregulated, for example, during malignant transformation. Non-limiting examples of ligands recognized by NKG2D include, but are not limited to, MICA, MICB, ULBP1, ULBP2, ULBP3, ULBP4, ULBP5, and ULBP6, as well as other molecules expressed on target cells that control the cytolytic or cytotoxic function of NK cells. In some embodiments, T cells are engineered to express an extracellular domain that binds to one or more tumor ligands and activates T lymphocytes. For example, in some embodiments, T cells are engineered to express the NKG2D receptor as a binding/activating moiety. In some embodiments, the engineered cells disclosed herein are engineered to express another member of the NKG2 family, such as NKG2A, NKG2C, and/or NKG2E. In some embodiments, combinations of such receptors are engineered. Moreover, in some embodiments, other receptors are expressed, such as killer cell immunoglobulin-like receptors (KIRs).

[00157] In several embodiments, the cells are engineered to express a cytotoxic receptor complex comprising full-length NKG2D as an extracellular component for recognizing ligands on the surface of tumor cells (e.g., liver cells). In one embodiment, the full-length NKG2D is characterized by the nucleic acid sequence of SEQ ID NO: 27. In several embodiments, the full-length NKG2D or a functional fragment thereof is human NKG2D.

[00158] In some embodiments, the cells are engineered to express a cytotoxic receptor complex comprising a functional fragment of NKG2D as an extracellular component for recognizing ligands on the surface of tumor cells or other diseased cells. In one embodiment, the functional fragment of NKG2D is characterized by the nucleic acid sequence of SEQ ID NO: 25. In some embodiments, the NKG2D fragment is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% homologous to full-length wild-type NKG2D. In some embodiments, the fragment may have one or more additional mutations in SEQ ID NO: 25, but retains or, in some embodiments, has enhanced ligand-binding function. In several embodiments, a functional fragment of NKG2D comprises the amino acid sequence of SEQ ID NO: 26. In several embodiments, the NKG2D fragment is provided as a dimer, trimer, or other concatameric format, wherein such embodiments provide for enhanced ligand-binding activity. In several embodiments, the sequence encoding the NKG2D fragment is optionally fully or partially codon-optimized. In one embodiment, the sequence encoding the codon-optimized fragment of NKG2D comprises the sequence of SEQ ID NO: 28. Preferably, according to several embodiments, the functional fragment lacks its natural transmembrane or intracellular domains, but retains its ability to bind NKG2D ligands, as well as to transduce activation signals upon ligand binding. An additional advantage of such fragments is that expression of DAP10 is not required for localization of NKG2D to the cell membrane. Thus, in several embodiments, the cytotoxic receptor complex encoded by the polypeptides disclosed herein does not comprise DAP10. In several embodiments, immune cells, such as NK or T cells, are engineered to express one or more chimeric receptors that target CD19 and NGG2D ligand. Such cells, in several embodiments, also co-express mbIL15.

[00159] In several embodiments, the cytotoxic receptor complexes are configured to dimerize. The dimerization may include homodimers or heterodimers, depending on the embodiment. In several embodiments, the dimerization results in improved ligand recognition by the cytotoxic receptor complexes (and therefore NK cells expressing the receptor), resulting in reduced (or absent) unwanted toxic effects. In several embodiments, the cytotoxic receptor complexes utilize internal dimers or repeats of one or more component subunits. For example, in several embodiments, the cytotoxic receptor complexes may optionally comprise a first NKG2D extracellular domain associated with a second NKG2D extracellular domain and a transmembrane/signaling region (or a separate transmembrane region together with a separate signaling region).

[00160] In several embodiments, the various domains/subdomains are separated by a linker, such as a GS3 linker (SEQ ID NOs: 15 and 16, a nucleotide and a protein, respectively) (or a GSn linker) is used. Other linkers used in accordance with various embodiments disclosed herein include, but are not limited to, linkers encoded by sequences of SEQ ID NOs: 17, 19, 21, or 23. This allows for the separation of various constituent parts of the receptor complex along a polynucleotide, which can enhance the expression, stability, and/or functionality of the receptor complex.

Cytotoxic signaling complex

[00161] Some embodiments of the compositions and methods described herein relate to a chimeric receptor, such as a CD19-targeted chimeric receptor, that comprises a cytotoxic signaling complex. As disclosed herein, in accordance with several embodiments, the provided cytotoxic receptor complexes comprise one or more transmembrane and/or intracellular domains that initiate cytotoxic signaling cascades upon binding of the extracellular domain(s) to ligands on the surface of target cells. Certain embodiments disclosed herein relate to chimeric antigen receptor constructs wherein the tumor targeting domain (or CD19-targeting domain) is linked to a cytotoxic signaling complex.

[00162] In some embodiments, the cytotoxic signaling complex comprises at least one transmembrane domain, at least one costimulatory domain, and/or at least one signaling domain. In some embodiments, more than one component makes up the domain, such as a costimulatory domain may comprise two subdomains. Moreover, in some embodiments, a domain may perform multiple functions, such as a transmembrane domain may also perform a signaling function.

Transmembrane domains

[00163] Some embodiments of the compositions and methods described herein relate to a chimeric receptor, such as a CD19-directed chimeric receptor, that comprises a transmembrane domain. Some embodiments include a transmembrane domain from NKG2D or another transmembrane protein. In some embodiments in which a transmembrane domain is used, the portion of the transmembrane protein used retains at least a portion of its normal transmembrane domain.

[00164] However, in some embodiments, the transmembrane domain comprises at least a portion of CD8, a transmembrane glycoprotein typically expressed on both T cells and NK cells. In some embodiments, the transmembrane domain comprises CD8α. In some embodiments, the transmembrane domain is referred to as a "hinge." In some embodiments, the CD8α hinge has a nucleic acid sequence of SEQ ID NO: 1. In some embodiments, the CD8α hinge is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to a CD8α having a sequence of SEQ ID NO: 1. In some embodiments, the CD8α hinge comprises an amino acid sequence of SEQ ID NO: 2. In some embodiments, the CD8α may be truncated or modified such that it is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the sequence of SEQ ID NO: 2.

[00165] In some embodiments, the transmembrane domain comprises a CD8α transmembrane region. In some embodiments, the transmembrane domain of CD8α is characterized by the nucleic acid sequence of SEQ ID NO: 3. In some embodiments, the CD8α hinge is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to a CD8α characterized by the sequence of SEQ ID NO: 3. In some embodiments, the transmembrane domain of CD8α comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the CD8α hinge is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to a CD8α characterized by the sequence of SEQ ID NO: 4.

[00166] Taken together, in several embodiments, the CD8 hinge/transmembrane complex is encoded by the nucleic acid sequence of SEQ ID NO: 13. In several embodiments, the CD8 hinge/transmembrane complex is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the CD8 hinge/transmembrane complex characterized by the sequence of SEQ ID NO: 13. In several embodiments, the CD8 hinge/transmembrane complex comprises the amino acid sequence of SEQ ID NO: 14. In several embodiments, the hinge of the CD8 hinge/transmembrane complex is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the CD8 hinge/transmembrane complex characterized by the sequence of SEQ ID NO: 14.

[00167] In some embodiments, the transmembrane domain comprises a CD28 transmembrane domain or a fragment thereof. In some embodiments, the CD28 transmembrane domain comprises the amino acid sequence of SEQ ID NO: 30. In some embodiments, the hinge of the CD28 transmembrane domain complex is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to a CD28 transmembrane domain characterized by the sequence of SEQ ID NO: 30.

Costimulatory domains

[00168] Some embodiments of the compositions and methods described herein relate to a chimeric receptor, such as a CD19-directed chimeric receptor, that comprises a costimulatory domain. In addition to the various transmembrane domains and the signaling domain (and combination of transmembrane/signaling domains), additional coactivating molecules may be provided in some embodiments. These may be certain molecules that, for example, further enhance the activity of immune cells. In some embodiments, cytokines may be used. For example, but not limited to, certain interleukins such as IL-2 and/or IL-15 are used. In some embodiments, the immune cells for therapy are engineered to express such molecules in a secreted form. In further embodiments, such costimulatory domains are engineered to be membrane-bound, acting as autocrine stimulatory molecules (or even as paracrine stimulators of neighboring cells being delivered). In some embodiments, NK cells are engineered to express membrane-associated interleukin 15 (mbIL15). In such embodiments, expression of mbIL15 on NK enhances the cytotoxic effects of the engineered NK cell by increasing proliferation and/or survival time of the NK cells. In some embodiments, mbIL15 has the nucleic acid sequence of SEQ ID NO: 11. In some embodiments, mbIL15 may be truncated or modified such that it is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the sequence of SEQ ID NO: 11. In some embodiments, mbIL15 comprises the amino acid sequence of SEQ ID NO: 12. In some embodiments, mbIL15 is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to mbIL15 having the sequence of SEQ ID NO: 12.

[00169] In some embodiments, the CD19-directed chimeric receptor or engineered cytotoxic receptor complex is encoded by a polynucleotide that includes one or more cytosolic protease cleavage sites, such as a T2A cleavage site, a P2A cleavage site, an E2A cleavage site, and/or an F2A cleavage site. Such sites are recognized and cleaved by a cytosolic protease, which can result in the separation (and separate expression) of the various constituent parts of the receptor encoded by the polynucleotide. As a result, depending on the embodiment, the various constituent parts of the CD19-directed chimeric receptor or engineered cytotoxic receptor complex can be delivered to an NK cell or T cell in a single vector or using multiple vectors. Thus, as schematically shown in the figures, the construct may be encoded by a single polynucleotide, but also include a cleavage site such that the downstream elements of the constructs are expressed by cells as a separate protein (as is the case with IL-15 in some embodiments). In some embodiments, a T2A cleavage site is used. In some embodiments, the T2A cleavage site is characterized by the nucleic acid sequence of SEQ ID NO: 9. In some embodiments, the T2A cleavage site may be truncated or modified such that it is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the sequence of SEQ ID NO: 9. In some embodiments, the T2A cleavage site comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the T2A cleavage site is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the T2A cleavage site having the sequence of SEQ ID NO: 10.

Signaling domains

[00170] Some embodiments of the compositions and methods described herein relate to a chimeric receptor, such as a CD19-directed chimeric receptor, that comprises a signaling domain. For example, immune cells engineered in accordance with several embodiments disclosed herein can comprise at least one subunit of the CD3 T cell receptor complex (or a fragment thereof). In several embodiments, the signaling domain comprises the CD3 zeta subunit. In some embodiments, CD3-zeta is encoded by the nucleic acid sequence of SEQ ID NO: 7. In some embodiments, CD3-zeta may be truncated or modified such that it is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the CD3-zeta domain characterized by the sequence of SEQ ID NO: 7. In some embodiments, the CD3-zeta domain comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the CD3-zeta domain is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the CD3-zeta domain characterized by the sequence of SEQ ID NO: 8.

[00171] In some embodiments, unexpectedly, the enhancement of signaling is achieved by using multiple signaling domains whose activities are synergistic. For example, in some embodiments, the signaling domain further comprises an OX40 domain. In some embodiments, the OX40 domain is an intracellular signaling domain. In some embodiments, the OX40 intracellular signaling domain is characterized by the nucleic acid sequence of SEQ ID NO: 5. In some embodiments, the OX40 intracellular signaling domain may be truncated or modified such that it is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to OX40 characterized by the sequence of SEQ ID NO: 5. In some embodiments, the OX40 intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 16. In some embodiments, the OX40 intracellular signaling domain is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the intracellular signaling domain of OX40, characterized by the sequence of SEQ ID NO: 6. In some embodiments, OX40 is used as the only transmembrane/signaling domain in the construct, however, in some embodiments, OX40 can be used with one or more other domains. For example, in some embodiments, combinations of OX40 and CD3-zeta are used. As a further example, in some embodiments, combinations of CD28, OX40, 4-1BB and/or CD3-zeta are used.

[00172] In some embodiments, the signaling domain comprises a 4-1BB domain. In some embodiments, the 4-1BB domain is an intracellular signaling domain. In some embodiments, the 4-1BB intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 29. In some embodiments, the 4-1BB intracellular signaling domain is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the 4-1BB intracellular signaling domain having the sequence of SEQ ID NO: 29. In some embodiments, 4-1BB is used as the only transmembrane/signaling domain in the construct, however, in some embodiments, 4-1BB can be used with one or more other domains. For example, in some embodiments, combinations of 4-1BB and CD3-zeta are used. As a further example, in some embodiments, combinations of CD28, OX40, 4-1BB and/or CD3-zeta are used.

[00173] In some embodiments, the signaling domain comprises a CD28 domain. In some embodiments, the CD28 domain is an intracellular signaling domain. In some embodiments, the CD28 intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 31. In some embodiments, the CD28 intracellular signaling domain is truncated or modified and is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% homologous to the CD28 intracellular signaling domain having the sequence of SEQ ID NO: 31. In some embodiments, CD28 is used as the only transmembrane/signaling domain in the construct, however, in some embodiments, CD28 can be used with one or more other domains. For example, in some embodiments, combinations of CD28 and CD3-zeta are used. As a further example, in some embodiments, combinations of CD28, OX40, 4-1BB and/or CD3-zeta are used.

Constructs of the cytotoxic receptor complex

[00174] Some embodiments of the compositions and methods described herein relate to a chimeric receptor, such as a CD19-directed chimeric receptor, that comprises a cytotoxic receptor complex or a cytotoxic receptor complex construct. As described above, various cytotoxic receptor complexes (also referred to as cytotoxic receptors) are provided herein. Expression of these complexes in immune cells, such as T cells and/or NK cells, allows for the targeting and destruction of specific target cells, such as cancer cells. Non-limiting examples of such cytotoxic receptor complexes are discussed in more detail below.

Constructs of the chimeric antigen receptor cytotoxic receptor complex

[00175] In several embodiments, provided herein are various cytotoxic receptor complexes (also referred to as cytotoxic receptors) with the general structure of a chimeric antigen receptor. Figures 1A, 1B, and 2 are schematic diagrams of non-limiting constructs that include a moiety of CD19 that binds to tumor antigens or tumor-associated antigens expressed on the surface of cancer cells and activates engineered cells expressing the chimeric antigen receptor. As shown in the figures, several embodiments of the chimeric receptor include an anti-CD19 moiety, a CD8a hinge domain, an Ig4 SH domain (or hinge), a CD8a transmembrane domain, a CD28 transmembrane domain, an OX40 domain, a 4-1BB domain, a CD28 domain, a CD3ζ ITAM domain or subdomain, a CD3-zeta domain, an NKp80 domain, a CD16 IC domain, a 2A cleavage site, and a membrane-bound IL-15 domain (although, as noted above, several embodiments use soluble IL-15). In several embodiments, the binding and activation functions are engineered to be performed by separate domains. Several embodiments relate to complexes with more than one anti-CD19 moiety or other binding/activating moiety. In some embodiments, the binding/activating moiety targets markers other than CD19, such as a cancer antigen target described herein. In some embodiments, the overall structure of the chimeric antigen receptor construct includes a hinge and/or transmembrane domain. In some embodiments, these may be accomplished by a single domain, or in some embodiments, multiple subdomains may be used. The receptor complex further comprises a signaling domain that transduces signals upon binding of the homing moiety to the target cell, ultimately resulting in cytotoxic effects on the target cell. In some embodiments, the complex further comprises a costimulatory domain that acts synergistically, in some embodiments, to enhance the function of the signaling domain. Expression of these complexes in immune cells such as T cells and/or NK cells allows for the targeting and destruction of specific target cells such as cancer cells that express CD19. Some of these receptor complexes contain an extracellular domain containing an anti-CD19 moiety or a CD19-binding moiety that binds CD19 on the surface of target cells and activates the engineered cells. The CD3zeta ITAM subdomain can act cooperatively as a signaling domain. An IL-15 domain, such as that of mbIL-15, can act as a costimulatory domain. An IL-15 domain, such as that of mbIL-15, can render immune cells (e.g., NK or T cells) expressing it particularly effective against tumor target cells. It should be appreciated that the IL-15 domain, such as the mbIL-15 domain, may, in accordance with several embodiments, be encoded in a separate construct. Additionally, each of the components may be encoded in one or more separate constructs. In some embodiments, the cytotoxic receptor or CD19-directed receptor comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or within a range defined by any two of the aforementioned percentages, identical to the sequence of SEQ ID NO: 34.

[00176] In one embodiment, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an κCD19/CD8 hinge-CD8TM/OX40/CD3-zeta fragment (see Figure 1A, CD19-1a). The polynucleotide comprises or consists of the κCD19 fragment, the CD8a hinge, the CD8a transmembrane domain, the OX40 domain, and the CD3-zeta domain described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more of the SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00177] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge-CD8TM/OX40/CD3-zeta/2A/mIL-15 (see Figure 1A, CD19-1b). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD8a transmembrane domain, an OX40 domain, a CD3-zeta domain, a cleavage site 2A, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00178] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/Ig4SH-CD8TM/4-1BB/CD3-zeta fragment (see Figure 1A, CD19-2a). The polynucleotide comprises or consists of an α-CD19 fragment, an Ig4 SH domain, a CD8a transmembrane domain, a 4-1BB domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00179] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/Ig4SH-CD8TM/4-1BB/CD3-zeta/2A/mIL-15 fragment (see Figure 1A, CD19-2b). The polynucleotide comprises or consists of an anti-CD19 fragment, an Ig4 SH domain, a CD8a transmembrane domain, a 4-1BB domain, a CD3-zeta domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00180] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19 fragment/CD8-CD28TM/CD28/CD3-zeta hinge (see Figure 1A, CD19-3a). The polynucleotide comprises or consists of an α-CD19 fragment, a CD8a hinge, a CD28 transmembrane domain, a CD28 domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more of the SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00181] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8-CD28TM/CD28/CD3-zeta/2A/mIL-15 hinge (see Figure 1A, CD19-3b). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD28 transmembrane domain, a CD28 domain, a CD3-zeta domain, a cleavage site 2A, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00182] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/Ig4SH-CD28TM/CD28/CD3-zeta fragment (see Figure 1A, CD19-4a). The polynucleotide comprises or consists of an α-CD19 fragment, an Ig4 SH domain, a CD28 transmembrane domain, a CD28 domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00183] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/Ig4SH-CD28TM/CD28/CD3-zeta/2A/mIL-15 fragment (see Figure 1A, CD19-4b). The polynucleotide comprises or consists of an α-CD19 fragment, an Ig4 SH domain, a CD28 transmembrane domain, a CD28 domain, a CD3-zeta domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00184] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/Ig4SH-CD8TM/OX40/CD3-zeta fragment (see Figure 2A, CD19-5a). The polynucleotide comprises or consists of an α-CD19 fragment, an Ig4 SH domain, a CD8a transmembrane domain, an OX40 domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more of the SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00185] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/Ig4SH-CD8TM/OX40/CD3-zeta/2A/mIL-15 fragment (see Figure 2A, CD19-5b). The polynucleotide comprises or consists of an α-CD19 fragment, an Ig4 SH domain, a CD8a transmembrane domain, an OX40 domain, a CD3-zeta domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00186] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19 fragment/CD8-CD3αTM/CD28/CD3-zeta hinge (see Figure 1B, CD19-6a). The polynucleotide comprises or consists of an α-CD19 fragment, a CD8a hinge, a CD3α transmembrane domain, a CD28 domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more of the sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00187] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8-CD3αTM hinge/CD28/CD3-zeta/2A/mIL-15 (see Figure 1B, CD19-6b). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD3α transmembrane domain, a CD28 domain, a CD3-zeta domain, a cleavage site 2A, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00188] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge-CD28TM/CD28/4-1BB/CD3-zeta (see Figure 1B, CD19-7a). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD28 transmembrane domain, a CD28 domain, a 4-1BB domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00189] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge-CD28TM/CD28/4-1BB/CD3-zeta/2A/mIL-15 (see Figure 1B, CD19-7b). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD28 transmembrane domain, a CD28 domain, a 4-1BB domain, a CD3-zeta domain, a cleavage site 2A, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00190] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an κCD19 fragment/CD8alpha hinge/TM CD8alpha/4-1BB/CD3zeta (see Figure 2, CD19-8a). The polynucleotide comprises or consists of an κCD19 fragment, a CD8a hinge, a CD8a transmembrane domain, a 4-1BB domain, and a CD3zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00191] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/CD8alpha hinge/TM CD8alpha/4-1BB/CD3zeta/2A/mIL-15 fragment (see Figure 2, CD19-8b). The polynucleotide comprises or consists of an α-CD19 fragment, a CD8a hinge, a CD8a transmembrane domain, a 4-1BB domain, a CD3zeta domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00192] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an κCD19 fragment/CD8alpha hinge/TM CD3/4-1BB/CD3zeta (see Figure 2, CD19-39_5a). The polynucleotide comprises or consists of an κCD19 fragment, a CD8a hinge, a CD3 transmembrane domain, a 4-1BB domain, and a CD3zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00193] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19/CD8alpha hinge/TM CD3/4-1BB/CD3-zeta/2A/mIL-15 fragment (see Figure 2, CD19-39_5b). The polynucleotide comprises or consists of an α-CD19 fragment, a CD8a hinge, a CD8a transmembrane domain, a 4-1BB domain, a CD3-zeta domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00194] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19 fragment/CD8alpha hinge/TM CD3/4-1BB/NKp80 (see Figure 2, CD19-39_6a). The polynucleotide comprises or consists of an α-CD19 fragment, a CD8a hinge, a CD3 transmembrane domain, a 4-1BB domain, and an NKp80 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In several embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more of the SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more of the sequences of SEQ ID NOs described herein.

[00195] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8alpha hinge/TM CD3/4-1BB/NKp80/2A/mIL-15 (see Figure 2, CD19-39_6b). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD8a transmembrane domain, a 4-1BB domain, an NKp80 domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00196] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an α-CD19 fragment/CD8alpha hinge/CD3TM/CD16 intracellular domain/4-1BB (see Figure 2, CD19-39_10a). The polynucleotide comprises or consists of an α-CD19 fragment, a CD8a hinge, a CD3 transmembrane domain, a CD16 intracellular domain, and a 4-1BB domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00197] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8alpha hinge/TM CD3/CD16/4-1BB/2A/mIL-15 (see Figure 2, CD19-39_10b). The polynucleotide comprises or consists of an anti-CD19 fragment, a CD8a hinge, a CD8a transmembrane domain, a CD16 intracellular domain, a 4-1BB domain, a 2A cleavage site, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00198] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/NKG2D extracellular domain/CD8-CD8TM hinge/OX40/CD3-zeta (see Figure 2, CD19/NKG2D-1a). The polynucleotide comprises or consists of an anti-CD19 fragment, an NKG2D extracellular domain (either full length or a fragment), a CD8a hinge, a CD8a transmembrane domain, an OX40 domain, and a CD3-zeta domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00199] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/NKG2D EC domain/CD8-CD8TM hinge/OX40/CD3-zeta/2A/mIL-15 (see Figure 2, CD19/NKG2D-1b). The polynucleotide comprises or consists of an anti-CD19 fragment, an NKG2D extracellular domain (either full length or a fragment), a CD8a hinge, a CD8a transmembrane domain, an OX40 domain, a CD3-zeta domain, a cleavage site 2A, and an mIL-15 domain as described herein. In several embodiments, the receptor complex is encoded by a nucleic acid molecule comprising a sequence derived from a combination of the sequences disclosed herein or comprises an amino acid sequence derived from a combination of the sequences disclosed herein. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence according to one or more sequences of SEQ ID NOs described herein, such as those included herein as exemplary constituent parts. In some embodiments, the coding nucleic acid sequence or amino acid sequence comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to a sequence resulting from a combination of one or more sequences of SEQ ID NOs described herein.

[00200] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8TM/4-1BB/CD3-zeta/mbIL15 fragment (see Figure 3A, NK19). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a 4-1BB domain, and a CD3-zeta domain. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 85. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 85. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 86. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 86. A schematically depicted and useful NK19 construct lacking the mbIL15 domain is provided (Figure 3A, NK19 variant)

[00201] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3A, NK19-1a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, an OX40 domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3A, NK19-1b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, an OX40 domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 59. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 59. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 60. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 60. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00202] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD28TM/CD28/CD3-zeta fragment (see Figure 3A, NK19-2a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD28 transmembrane domain, a CD28 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3A, NK19-2b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD28 transmembrane domain, a CD28 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 61. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 61. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 62. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 62. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00203] In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge/CD8aTM/ICOS/CD3-zeta (see Figure 3A, NK19-3a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, an inducible costimulator (ICOS) signaling domain, and a CD3-zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3A, NK19-3b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, an inducible costimulator signaling (ICOS) domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 63. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 63. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 64. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 64. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00204] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/CD28/4-1BB/CD3-zeta fragment (see Figure 3A, NK19-4a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD28 signaling domain, a 4-1BB signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3A, NK19-4b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD28 signaling domain, a 4-1BB signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 65. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 65. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 66. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 66. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00205] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/NKG2DTM/OX40/CD3-zeta fragment (see Figure 3B, NK19-5a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, an NKG2D transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3B, NK19-5b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, an NKG2D transmembrane domain, an OX40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 67. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 67. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 68. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 68. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00206] In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge/CD8aTM/CD40/CD3-zeta (see Figure 3B, NK19-6a). The polynucleotide comprises or consists of an anti-CD19 scFv heavy chain variable region, a CD8a hinge, a CD8a transmembrane domain, a CD40 signaling domain, and a CD3-zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3B, NK19-6b). In such embodiments, the polynucleotide comprises or consists of the heavy chain variable region of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 69. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 69. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 70. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 70. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00207] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/OX40/CD3-zeta/2A/EGFRt fragment (see Figure 3B, NK19-7a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and a truncated version of the epidermal growth factor receptor (EGFRt). In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3B, NK19-7b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, a CD3-zeta domain, a cleavage site 2A, a truncated version of the epidermal growth factor receptor (EGFRt), an additional cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 71. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 71. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 72. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 72. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00208] In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge/CD8aTM/CD40/CD3-zeta (see Figure 3B, NK19-8a). The polynucleotide comprises or consists of an anti-CD19 light chain variable region of a CD8a hinge, a CD8a transmembrane domain, a CD40 signaling domain, and a CD3-zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3B, NK19-7b). In such embodiments, the polynucleotide comprises or consists of the light chain variable region of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 73. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 73. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 74. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 74. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00209] In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19 fragment/CD8 hinge/CD8aTM/CD40/CD3-zeta (see Figure 3B, NK19-8a). The polynucleotide comprises or consists of an anti-CD19 light chain variable region of a CD8a hinge, a CD8a transmembrane domain, a CD40 signaling domain, and a CD3-zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3B, NK19-7b). In such embodiments, the polynucleotide comprises or consists of the light chain variable region of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 73. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 73. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 74. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 74. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00210] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/CD27/CD3-zeta fragment (see Figure 3C, NK19-9a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD27 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3C, NK19-9b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD27 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 75. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 75. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 76. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 76. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00211] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/CD70/CD3-zeta fragment (see Figure 3C, NK19-10a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD70 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3C, NK19-10b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD70 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 77. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 77. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 78. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 78. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00212] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/CD161/CD3-zeta fragment (see Figure 3C, NK19-11a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD161 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3C, NK19-11b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD161 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 79. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 79. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 80. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 80. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00213] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/CD40L/CD3-zeta fragment (see Figure 3C, NK19-12a). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD40L signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3C, NK19-12b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD40L signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 81. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 81. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 82. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 82. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00214] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is an anti-CD19/CD8 hinge/CD8aTM/CD44/CD3-zeta fragment (see Figure 3C, NK19-13). The polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD44 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3C, NK19-13b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a CD44 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 83. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 83. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 84. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 84. In several embodiments, the anti-CD19 scFv does not comprise a Flag tag.

[00215] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3D, NK19H-1a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a first humanized heavy chain (L1/H1), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3D, NK19H-1b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a first humanized heavy chain (L1/H1), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 160. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 160. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 161. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 161.

[00216] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3D, NK19H-2a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a first humanized heavy chain (L2/H1), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3D, NK19H-2b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized, comprises a second humanized light chain and a first humanized heavy chain (L2/H1), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 162. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 162. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 163. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 162.

[00217] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3D, NK19H-3a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a first humanized heavy chain (L3/H1), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3D, NK19H-3b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a first humanized heavy chain (L3/H1), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 164. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 164. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 165. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 165.

[00218] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3D, NK19H-4a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a second humanized heavy chain (L1/H2), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3D, NK19H-4b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a second humanized heavy chain (L1/H2), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 166. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 166. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 167. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 167.

[00219] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3E, NK19H-5a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a second humanized heavy chain (L2/H2), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3E, NK19H-5b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a second humanized heavy chain (L2/H2), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 168. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 168. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 169. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 169.

[00220] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3E, NK19H-6a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a second humanized heavy chain (L3/H2), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3E, NK19H-6b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a second humanized heavy chain (L3/H2), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 170. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 170. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 171. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 171.

[00221] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3E, NK19H-7a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a third humanized heavy chain (L1/H3), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3E, NK19H-7b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a third humanized heavy chain (L1/H3), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, a truncated version of the epidermal growth factor receptor (EGFRt), an additional cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 172. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 172. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 173. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 174.

[00222] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3E, NK19H-8a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a third humanized heavy chain (L2/H3), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3E, NKH19-8b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a third humanized heavy chain (L2/H3), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 174. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 174. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 175. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 175.

[00223] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3E, NK19H-9a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a third humanized heavy chain (L3/H3), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3E, NKH19-9b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a third humanized heavy chain (L3/H3), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 176. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 176. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 177. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 177.

[00224] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3E, NKH19-10a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a fourth humanized heavy chain (L1/H4), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3E, NK19H-10b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a first humanized light chain and a fourth humanized heavy chain (L1/H4), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 178. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 178. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 179. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 179.

[00225] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3F, NK19H-11a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a fourth humanized heavy chain (L2/H4), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3F, NK19H-11b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a fourth humanized heavy chain (L2/H4), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 180. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 180. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 181. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 181.

[00226] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8TM/OX40/CD3-zeta (see Figure 3F, NK19H-12a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a fourth humanized heavy chain (L3/H4), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3F, NK19H-12b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a fourth humanized heavy chain (L3/H4), and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a 2A cleavage site, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 182. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 182. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 183. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 183.

[00227] In several embodiments, a polynucleotide encoding a chimeric antigen receptor is provided that comprises a humanized Flag-tagged fragment of CD19 and multiple costimulatory domains. For example, the schematic structure is a fragment of CD19/transmembrane domain/costimulatory domain 1/costimulatory domain 2/costimulatory domain 3/signaling domain. The costimulatory domains vary in order depending on the embodiment. For example, in some embodiments, the costimulatory domains ("CSDs") can be arranged as: CSD1/CSD2, CSD2/CSD1, CSD1/CSD2/CSD3, CSD1/CSD2/CSD3, CSD3/CSD2/CSD1, etc. In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8aTM/CD44/OX40/CD27/CD3-zeta (see Figure 3F, NK19H-13a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, a CD44 costimulatory domain, an OX40 costimulatory domain, a CD27 costimulatory domain, and a CD3-zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3F, NK19H-13b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv that has been humanized and comprises a Flag tag, a CD8a hinge, a CD8a transmembrane domain, a CD44 costimulatory domain, an OX40 costimulatory domain, a CD27 costimulatory domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein.

[00228] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3G, NK19H-NF-1a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a first humanized heavy chain (L1/H1), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3G, NK19H-NF-1b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a first humanized heavy chain (L1/H1), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 184. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 184. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 185. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 185.

[00229] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3G, NK19H-NF-2a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a second humanized light chain and a first humanized heavy chain (L2/H1), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3G, NK19H-NF-2b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a first humanized heavy chain (L2/H1), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 186. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 186. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 187. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 187.

[00230] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3G, NK19H-NF-3a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a first humanized heavy chain (L3/H1), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3G, NK19H-NF-3b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a first humanized heavy chain (L3/H1), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 188. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 188. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 189. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 189.

[00231] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3G, NK19H-NF-4a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a second humanized heavy chain (L1/H2), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3G, NK19H-NF-4b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a second humanized heavy chain (L1/H2), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 190. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 190. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 191. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 191.

[00232] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3H, NK19H-NF-5a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a second humanized light chain and a second humanized heavy chain (L2/H2), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3H, NK19H-NF-5b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a second humanized light chain and a second humanized heavy chain (L2/H2), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 192. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 192. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 193. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 193.

[00233] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3H, NK19H-NF-6a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a second humanized heavy chain (L3/H2), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3H, NK19H-NF-6b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv that has been humanized and comprises a third humanized light chain and a second humanized heavy chain (L3/H2), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 194. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 194. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 195. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 195.

[00234] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3G, NK19H-NF-7a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a third humanized heavy chain (L1/H3), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3H, NK19H-NF-7b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a third humanized heavy chain (L1/H3), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 196. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 196. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 197. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 197.

[00235] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3H, NK19H-NF-8a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a second humanized light chain and a third humanized heavy chain (L2/H3), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3H, NKH19-NF-8b). In such embodiments, the polynucleotide comprises a light chain variable region of an anti-CD19 scFv that has been humanized and comprises or consists of a second humanized light chain and a third humanized heavy chain (L2/H3), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 198. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 198. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 199. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 199.

[00236] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3H, NK19H-NF-9a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a third humanized heavy chain (L3/H3), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3H, NKH19-NF-9b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a third humanized heavy chain (L3/H3), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 200. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 200. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 201. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 201.

[00237] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3H, NKH19-NF-10a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a fourth humanized heavy chain (L1/H4), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3H, NK19H-NF-10b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a first humanized light chain and a fourth humanized heavy chain (L1/H4), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 202. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 202. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 203. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 203.

[00238] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3I, NK19H-NF-11a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a second humanized light chain and a fourth humanized heavy chain (L2/H4), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3I, NK19H-NF-11b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv that has been humanized and comprises a second humanized light chain and a fourth humanized heavy chain (L2/H4), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 204. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 204. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 205. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 205.

[00239] In several embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized anti-CD19/CD8 hinge/CD8TM/OX40/CD3-zeta fragment (see Figure 3I NK19H-12a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a fourth humanized heavy chain (L3/H4), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain. In several embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3I, NK19H-NF-12b). In such embodiments, the polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a third humanized light chain and a fourth humanized heavy chain (L3/H4), a CD8a hinge, a CD8a transmembrane domain, an OX40 signaling domain, and a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein. In some embodiments, the receptor complex is encoded by a nucleic acid molecule having the sequence of SEQ ID NO: 206. In some embodiments, the nucleic acid sequence encoding the chimeric NK19 antigen receptor comprises a sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 206. In some embodiments, the chimeric receptor comprises the amino acid sequence of SEQ ID NO: 207. In some embodiments, the chimeric NK19 antigen receptor comprises an amino acid sequence that has at least about 90%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity, homology, and/or functional equivalence to the sequence of SEQ ID NO: 207.

[00240] In several embodiments, a polynucleotide encoding a chimeric antigen receptor is provided that comprises a humanized Flag-tagged fragment of CD19 and multiple costimulatory domains. For example, the schematic structure is a fragment of CD19/transmembrane domain/costimulatory domain 1/costimulatory domain 2/costimulatory domain 3/signaling domain. The costimulatory domains vary in order depending on the embodiment. For example, in some embodiments, the costimulatory domains ("CSDs") can be arranged as: CSD1/CSD2, CSD2/CSD1, CSD1/CSD2/CSD3, CSD1/CSD2/CSD3, CSD3/CSD2/CSD1, etc. In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8aTM/CD44/OX40/CD27/CD3zeta (see Figure 3I, NK19H-NF-13a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, a CD44 costimulatory domain, an OX40 costimulatory domain, a CD27 costimulatory domain, and a CD3zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3I, NK19H-NF-13b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv that has been humanized and comprises a Flag tag, a CD8a hinge, a CD8a transmembrane domain, a CD44 costimulatory domain, an OX40 costimulatory domain, a CD27 costimulatory domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein.

[00241] In several embodiments, a polynucleotide encoding a chimeric antigen receptor is provided that comprises a humanized Flag-tagged fragment of CD19 and multiple costimulatory domains. For example, the schematic structure is a fragment of CD19/transmembrane domain/costimulatory domain 1/costimulatory domain 2/costimulatory domain 3/signaling domain. The costimulatory domains vary in order depending on the embodiment. For example, in some embodiments, the costimulatory domains ("CSDs") can be arranged as: CSD1/CSD2, CSD2/CSD1, CSD1/CSD2/CSD3, CSD1/CSD2/CSD3, CSD3/CSD2/CSD1, etc. In some embodiments, a polynucleotide encoding a chimeric antigen receptor complex is provided that is a humanized fragment with an anti-CD19 Flag tag/CD8 hinge/CD8aTM/CD44/OX40/CD27/CD3-zeta (see Figure 3F, NK19H-13a). The polynucleotide comprises an anti-CD19 scFv that has been humanized and comprises or consists of a Flag tag, a CD8a hinge, a CD8a transmembrane domain, a CD44 costimulatory domain, an OX40 costimulatory domain, a CD27 costimulatory domain, and a CD3-zeta domain. In some embodiments, the chimeric antigen receptor further comprises mbIL15 (see Figure 3F, NK19H-13b). In such embodiments, the polynucleotide comprises or consists of an anti-CD19 scFv that has been humanized and comprises a Flag tag, a CD8a hinge, a CD8a transmembrane domain, a CD44 costimulatory domain, an OX40 costimulatory domain, a CD27 costimulatory domain, a CD3-zeta domain, a cleavage site 2A, and an mbIL-15 domain as described herein.

[00242] It should be appreciated that for any receptor construct described herein, certain sequence variability, extensions and/or truncations of the disclosed sequences may occur when linking the sequences as a result of, for example, facilitating or ensuring cloning efficiency (e.g., to create a restriction site).

Treatment methods

[00243] Some embodiments relate to a method of treating, ameliorating, inhibiting, or preventing cancer using a cell or immune cell comprising a chimeric receptor, such as a CD19-directed chimeric receptor. In some embodiments, the method comprises treating or preventing cancer. In some embodiments, the method comprises administering a therapeutically effective amount of immune cells expressing a CD19-directed chimeric receptor described herein. Examples of types of cancer that can be treated as such are described herein.

[00244] In certain embodiments, treating a subject with the genetically engineered cell(s) described herein achieves one, two, three, four, or more of the following effects, including, for example: (i) reducing or alleviating the severity of a disease or an associated symptom; (ii) reducing the duration of a symptom associated with the disease; (iii) protecting against progression of the disease or an associated symptom; (iv) regression of the disease or an associated symptom; (v) protecting against the development or occurrence of a symptom associated with the disease; (vi) protecting against the recurrence of a symptom associated with the disease; (vii) reducing the frequency of hospitalization of the subject; (viii) reducing the duration of hospitalization; (ix) increasing the survival of the subject with the disease; (x) reducing the number of symptoms associated with the disease; (xi) enhancing, improving, maintaining, complementing, or increasing the prophylactic or therapeutic effect(s) of another therapy. Administration may be by a variety of routes, including but not limited to intravenous, intraarterial, subcutaneous, intramuscular, intrahepatic, intraperitoneal, and/or local delivery to the affected tissue.

Administration and Dosage

[00245] Additionally provided herein are methods of treating a subject having a cancer comprising administering to the subject a composition comprising immune cells (such as NK and/or T cells) engineered to express a cytotoxic receptor complex disclosed herein. For example, some embodiments of the compositions and methods described herein relate to the use of a CD19-directed chimeric receptor or the use of cells expressing a CD19-directed chimeric receptor to treat a patient having a cancer. Also provided are methods of using such engineered immune cells to treat cancer.

[00246] In certain embodiments, treating a subject with the genetically engineered cell(s) described herein achieves one, two, three, four, or more of the following effects, including, for example: (i) reducing or alleviating the severity of a disease or an associated symptom; (ii) reducing the duration of a symptom associated with the disease; (iii) protecting against progression of the disease or an associated symptom; (iv) regression of the disease or an associated symptom; (v) protecting against the development or occurrence of a symptom associated with the disease; (vi) protecting against the recurrence of a symptom associated with the disease; (vii) reducing the frequency of hospitalization of the subject; (viii) reducing the duration of hospitalization; (ix) increasing the survival of the subject with the disease; (x) reducing the number of symptoms associated with the disease; (xi) enhancing, improving, maintaining, complementing, or increasing the prophylactic or therapeutic effect(s) of another therapy. Each of these comparisons is made relative to, for example, another disease therapy that includes cellular immunotherapy of the disease using cells that do not express the constructs disclosed herein.

[00247] Administration can be by a variety of routes, including, but not limited to, intravenous, intraarterial, subcutaneous, intramuscular, intrahepatic, intraperitoneal, and/or local delivery to the affected tissue. Doses of immune cells, such as NK and/or T cells, can be readily determined for a given subject based on their body weight, the type of disease and condition, and the desired aggressiveness of treatment, but, depending on embodiments, are in the range of about 10 ⁵ cells per kg to about 10 ¹² cells per kg (e.g., 10 ⁵ -10 ⁷ , 10 ⁷ -10 ¹⁰ , 10 ¹⁰ -10 ¹² and overlapping ranges of these values). In one embodiment, a dose escalation schedule is used. In some embodiments, immune cells such as NK and/or T cells are administered in a dose range of, for example, about 1 x 10 ⁶ cells/kg to about 1 x 10 ⁸ cells/kg. In some embodiments, the dose is in the range of about 2 x 10 ⁵ cells/kg to about 2 x 10 ⁸ cells/kg, including about 2 x 10 ⁶ and 2 x 10 ⁷ cells/kg. In some embodiments, the dose is determined by the maximum number of viable engineered cells at the time of dosing. For example, in some embodiments, a single dose contains a maximum of about 2 x 10 ⁵ to about 2 x 10 ⁹ viable engineered cells, including about 2 x 10 ⁶ , about 2 x 10 ⁷ , or about 2 x 10 ⁸ viable engineered cells. Depending on the embodiment, various types of cancer can be treated. In some embodiments, hepatocellular carcinoma is treated. Additional embodiments provided herein include treating or preventing the following non-limiting examples of cancer types, including but not limited to acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, Kaposi's sarcoma, lymphoma, gastrointestinal cancer, appendix cancer, central nervous system cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors (including but not limited to astrocytomas, spinal cord tumors, brainstem glioma, glioblastoma, craniopharyngioma, ependymoblastoma, ependymoma, medulloblastoma, medulloepithelioma), breast cancer, bronchial tumors, Burkitt's lymphoma, cervical cancer, colon cancer, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), Chronic myeloproliferative disorders, ductal carcinoma, endometrial cancer, esophageal cancer, gastric cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell leukemia, renal cell carcinoma, leukemia, oral cavity cancer, nasopharyngeal cancer, liver cancer, lung cancer (including but not limited to non-small cell lung cancer (NSCLC) and small cell lung cancer), pancreatic cancer, colorectal cancer, lymphoma, melanoma, eye cancer, ovarian cancer, pancreatic cancer, prostate cancer, pituitary cancer, uterine cancer, and vaginal cancer.

[00248] In some embodiments, provided herein are also nucleic acid and amino acid sequences that have at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% (and ranges therebetween) sequence identity or homology to the corresponding nucleic acid or amino acid sequences of SEQ ID NOs: 1-207 (or combinations of two or more of SEQ ID NOs: 1-207), and that also exhibit one or more functions compared to the corresponding sequences of SEQ ID NOs: 1-207 (or combinations of two or more of SEQ ID NOs: 1-207), including but not limited to (i) enhancing proliferation, (ii) enhancing activation, (iii) enhancing cytotoxic activity against cells presenting ligands to which the NK cells bearing receptors encoded by nucleic acid and amino acid sequences, (iv) enhanced homing to tumor or infected sites, (v) decreased non-targeted cytotoxic effects, (vi) enhanced secretion of immunostimulatory cytokines and chemokines (including but not limited to IFNg, TNFa, IL-22, CCL3, CCL4, and CCL5), (vii) enhanced ability to stimulate additional innate and adaptive immune responses, and (viii) combinations thereof.

[00249] Additionally, several embodiments provide amino acid sequences that correspond to any of the nucleic acids disclosed herein, taking into account the degeneracy of the nucleic acid code. Moreover, those sequences (whether a nucleic acid sequence or an amino acid sequence) that differ from those explicitly disclosed herein, but have functional similarity or equivalence, are also intended to be within the scope of the present invention. The foregoing includes mutants, truncations, substitutions, or other types of modifications.

[00250] In some embodiments, polynucleotides encoding the disclosed cytotoxic receptor complexes or CD19-directed chimeric receptors are mRNA. In some embodiments, the polynucleotide is DNA. In some embodiments, the polynucleotide is operably linked to at least one expression regulatory element of a cytotoxic receptor complex.

[00251] According to several embodiments, a vector is further provided comprising a polynucleotide encoding any of the polynucleotides provided herein, wherein the polynucleotides are optionally operably linked to at least one cytotoxic receptor complex expression regulatory element. In several embodiments, the vector is a retrovirus.

[00252] Further provided herein are engineered immune cells (such as NK and/or T cells) comprising a polynucleotide, vector, or cytotoxic receptor complexes disclosed herein. Further provided herein are compositions comprising a mixture of engineered immune cells (such as NK cells and/or engineered T cells), each population comprising a polynucleotide, vector, or cytotoxic receptor complexes disclosed herein.

[00253] Doses of immune cells such as NK cells or T cells can be readily determined for a given subject based on their body weight, the type of disease and condition, and the desired aggressiveness of treatment, but depending on embodiments, they range from about 10⁵cells per kg up to approximately 10¹²cells per kg (e.g. 10⁵-10⁷, 10⁷-10¹⁰, 10¹⁰-10¹²and in overlapping ranges of these values). In one embodiment, a dose escalation schedule is used. In several embodiments, the NK cells are administered in a dose range of, for example, from about 1 x 10⁶cells/kg to approximately 1 x 10⁸cells/kg. Depending on the embodiment, various types of cancer or infectious diseases can be treated.

Types of cancer

[00254] Some embodiments of the compositions and methods described herein relate to administering immune cells comprising a chimeric receptor, such as a CD19-directed chimeric receptor, to a subject having a cancer. Various embodiments provided herein include treating or preventing the following non-limiting examples of cancer types. Examples of cancer include but are not limited to acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, Kaposi's sarcoma, lymphoma, gastrointestinal cancer, appendix cancer, central nervous system cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors (including but not limited to astrocytomas, spinal cord tumors, brainstem glioma, craniopharyngioma, ependymoblastoma, ependymoma, medulloblastoma, medulloepithelioma), breast cancer, bronchial tumors, Burkitt's lymphoma, cervical cancer, colon cancer, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorders, ductal carcinoma, endometrial cancer, cancer esophageal cancer, stomach cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell leukemia, renal cell carcinoma, leukemia, oral cavity cancer, nasopharyngeal cancer, liver cancer, lung cancer (including but not limited to non-small cell lung cancer (NSCLC) and small cell lung cancer), pancreatic cancer, colorectal cancer, lymphoma, melanoma, eye cancer, ovarian cancer, pancreatic cancer, prostate cancer, pituitary cancer, uterine cancer, and vaginal cancer.

Targets that are cancer antigens

[00255] Some embodiments of the compositions and methods described herein relate to immune cells that comprise a chimeric receptor that targets a cancer antigen. Non-limiting examples of target antigens include: CD5, CD19; CD123; CD22; CD30; CD171; CS1 (also referred to as CD2 subset 1, CRACC, SLAMF7, CD319, and 19A24); C-type lectin-like molecule-1 (CLL-1 or CLECL1); CD33; epidermal growth factor receptor variant III (EGFRviii); ganglioside G2 (GD2); ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(l-l)Cer); TNF receptor family member B-cell maturation antigen (BCMA); Tn antigen ((Tn Ag) or (GalNAca-Ser/Thr)); prostate-specific membrane antigen (PSMA); receptor tyrosine kinase-like orphan receptor 1 (ROR1); Fms-like tyrosine kinase 3 (FLT3); tumor-associated glycoprotein 72 (TAG72); CD38; CD44v6; glycosylated epitope of CD43 expressed on acute leukemia or lymphoma cells but not on hematopoietic progenitor cells, glycosylated epitope of CD43 expressed on non-hematopoietic cancer cells, carcinoembryonic antigen (CEA); epithelial cell adhesion molecule (EPCAM); B7H3 (CD276); KIT (CD117); interleukin-13 receptor alpha-2 subunit (IL-13Ra2 or CD213A2); mesothelin; interleukin 11 receptor alpha (IL-llRa); prostate stem cell antigen (PSCA); serine protease 21 (testisin or PRSS21); vascular endothelial growth factor receptor 2 (VEGFR2); Lewis system antigen (Y); CD24; platelet-derived growth factor receptor beta (PDGFR-beta); stage-specific embryonic antigen-4 (SSEA-4); CD20; folate receptor alpha (FRa or FR1); folate receptor beta (FRb); receptor tyrosine protein kinase ERBB2 (Her2/neu); cell surface-associated mucin 1 (MUC1); epidermal growth factor receptor (EGFR); neural cell adhesion molecule (NCAM); prostasis; prostatic acid phosphatase (PAP); elongation factor 2 mutated (ELF2M); ephrin B2; fibroblast activation protein (FAP) alpha subunit; insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX); proteasome (prosome, macropain) subunit beta-type 9 (LMP2); glycoprotein 100 (gp100); oncogene fusion protein of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl); tyrosinase; ephrin receptor type A 2 (EphA2); sialylated Lewis adhesion molecule (sLe); GM3 ganglioside (aNeu5Ac(2-3)bDClalp(l-4)bDGlcp(l-l)Cer); transglutaminase 5 (TGS5); high molecular weight melanoma-associated antigen (HMWMAA); O-acetylated GD2 ganglioside (OAcGD2); tumor endothelial marker 1 (TEM1/CD248); tumor endothelial marker-related protein 7 (TEM7R); claudin 6 (CLDN6); thyroid-stimulating hormone receptor (TSHR); G protein-coupled receptor class C group 5 member D (GPRC5D); chromosome X open reading frame 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK); polysialic acid; placenta-specific protein 1 (PLAC1); hexasaccharide moiety of glycoceramide globoH (GloboH); mammary gland differentiation antigen (NY-BR-1); uroplakin 2 (UPK2); hepatitis A virus cellular receptor 1 (HAVCR1); beta-3-adrenergic receptor (ADRB3); pannexin 3 (PANX3); G protein-coupled receptor 20 (GPR20); lymphocyte antigen complex 6, locus K 9 (LY6K); olfactory receptor 51E2 (OR51E2); TCR-gamma alternative reading frame protein (TARP); Wilms tumor protein (WT1); cancer-testis antigen 1 (NY-ES0-1); cancer-testis antigen 2 (LAGE-la); melanoma-associated antigen 1 (MAGE-A1); ETS 6 translocation variant located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); X antigen family 1A (XAGE1); angiopoietin-binding cell surface receptor 2 (Tie 2); melanoma cancer-testicular antigen-1 (MAD-CT-1); melanoma cancer-testicular antigen-2 (MAD-CT-2); Fos-related antigen 1; p53 tumor protein (p53); mutant p53; prostein; survivin; telomerase; prostate carcinoma tumor antigen-1 (PCT A-1 or galectin 8), melanoma antigen recognized by T cells 1 (MelanA or MARTI); mutant rat sarcoma protein (Ras); human telomerase; human telomerase reverse transcriptase (hTERT); translocation variant of sarcoma antigen with punctate breakpoints; melanoma inhibitor of apoptosis (ML-IAP); ERG (fusion gene of transmembrane serine protease 2 (TMPRSS2) and ETS); N-acetylglucosaminyltransferase V (NA17); Pax-3 family of paired box protein (PAX3); androgen receptor; cyclin B1; neuroblastoma homolog of avian myelocytomatosis virus v-myc oncogene (MYCN); Ras homolog family member C (RhoC); tyrosinase-related protein 2 (TRP-2); cytochrome P450 IB 1 (CYPIB 1); zinc finger protein-binding factor CCCTC-like (BORIS or regulator of imprinted regions paralog), squamous cell carcinoma antigen recognized by T cells 3 (SART3); Pax paired box family protein 5 (PAX5); proacrosin-binding protein sp32 (OY-TES1); lymphocyte-specific protein tyrosine kinase (LCK); kinase A anchor protein 4 (AKAP-4); synovial sarcoma X protein with point breaks 2 (SSX2); receptor for advanced glycation end products (RAGE-1); renal ubiquitin antigen 1 (RU1); renal ubiquitin antigen 2 (RU2); legumain; human papillomavirus E6 (HPV E6); human papillomavirus E7 (HPV E7); intestinal carboxylesterase; mutated heat shock protein 70-2 (mut hsp70-2); CD79a; CD79b; CD72; leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); IgA receptor Fc fragment (FCAR or CD89); leukocyte immunoglobulin-like receptor subfamily A, member 2 (LILRA2); CD300-like molecule family f member (CD300LF); C-type lectin family 12 member A (CLEC12A); bone marrow stromal cell antigen 2 (BST2); EGF-like, module-containing, mucin-like, hormone receptor-like protein 2 (EMR2); lymphocyte antigen 75 (LY75); glypican-3 (GPC3); Fc receptor-like protein 5 (FCRL5) and immunoglobulin lambda-like polypeptide 1 (IGLL1), MPL, biotin, c-MYC epitope tag, CD34, LAMP1 TROP2, GFR-alpha-4, CDH17, CDH6, NYBR1, CDH19, CD200R, Slea (CA19.9; sialylated Lewis antigen); fucosyl-GM1, PTK7, gpNMB, CDH1-CD324, DLL3, CD276/B7H3, IL1 lRa, IL13Ra2, CD179b-IGLll, TCR-gamma-delta, NKG2D, CD32 (FCGR2A), Tn ag, Tim1-/HVCR1, CSF2RA (GM-CSFR-alpha), TGF-beta-R2, Lews Ag, TCR beta1 chain, TCR beta2 chain, TCR gamma chain, TCR delta chain, FITC, luteinizing hormone receptor (LHR), follicle-stimulating hormone receptor (FSHR), gonadotropin receptor (CGHR or GR), CCR4, GD3, SLAMF6, SLAMF4, HIV1 envelope glycoprotein, HTLV1-Tax, CMV pp65, EBV-EBNA3c, KSHV K8.1, KSHV-gH, influenza A hemagglutinin (HA), GAD, PDL1, guanylyl cyclase C (GCC), desmoglein 3 autoantibody (Dsg3), desmoglein 1 autoantibody (Dsgl), HLA, HLA-A, HLA-A2, HLA-B, HLA-C, HLA-DP, HLA-DM, HLA-DOA, HLA-DOB, HLA-DQ, HLA-DR, HLA-G, IgE, CD99, Ras G12V, tissue factor 1 (TF1), AFP, GPRC5D, claudin 18.2 (CLD18A2 or CLDN18A.2)), P-glycoprotein, STEAP1, Livl, nectin-4, protein Cripto, gpA33, BST1/CD157, low-conductance chloride channel, and TNT antibody-recognized antigen.

[00256] Additionally, in several embodiments, an immune cell is provided that expresses a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH domain comprises a VH domain having at least 95% identity to the VH domain amino acid sequence set forth in SEQ ID NO: 33, the VL domain having at least 95% identity to the VL domain amino acid sequence set forth in SEQ ID NO: 32, a hinge and/or transmembrane domain, an intracellular signaling domain, wherein the intracellular signaling domain comprises an OX40 subdomain, and wherein the cell further expresses membrane-bound interleukin-15 (mbIL15). In some embodiments, the intracellular signaling domain further comprises a CD3-zeta subdomain. In some embodiments, the OX40 subdomain comprises the amino acid sequence of SEQ ID NO: 6 and the CD3-zeta subdomain comprises the amino acid sequence of SEQ ID NO: 7. In some embodiments, the hinge domain comprises a CD8a hinge domain. In some embodiments, the CD8a hinge domain comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, mbIL15 comprises the amino acid sequence of SEQ ID NO: 12. In some embodiments, the chimeric receptor further comprises an extracellular domain of an NKG2D receptor. In some embodiments, the extracellular domain of an NKG2D receptor comprises a functional fragment of NKG2D comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the immune cell is a natural killer (NK) cell. In some embodiments, the immune cell is a T cell. In some embodiments, such immune cells are administered to a subject according to a method of treating cancer or are otherwise used to treat cancer, such as in the preparation of a drug for treating cancer. In some embodiments, the cancer is acute lymphocytic leukemia.

[00257] In several embodiments, a polynucleotide encoding a CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH domain has at least 95% identity to the VH domain amino acid sequence set forth in SEQ ID NO: 33, the VL domain has at least 95% identity to the VL domain amino acid sequence set forth in SEQ ID NO: 32, a hinge and/or transmembrane domain, an intracellular signaling domain, wherein the intracellular signaling domain comprises an OX40 subdomain, and wherein the polynucleotide further encodes membrane-associated interleukin-15 (mbIL15). In several embodiments, the intracellular signaling domain further comprises a CD3-zeta subdomain. In some embodiments, the encoded OX40 subdomain comprises the amino acid sequence of SEQ ID NO: 16, the encoded CD3zeta subdomain comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the hinge domain comprises the CD8a hinge domain and comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the encoded mbIL15 comprises the amino acid sequence of SEQ ID NO: 12. In some embodiments, the chimeric receptor further comprises the extracellular domain of the NKG2D receptor. In some embodiments, the encoded extracellular domain of the NKG2D receptor comprises a functional fragment of NKG2D comprising the amino acid sequence of SEQ ID NO: 26.

[00258] Also provided herein is an immune cell that expresses a CD19-directed chimeric receptor comprising an extracellular fragment of CD19, a hinge and/or transmembrane domain, and an intracellular signaling domain. In some embodiments, the immune cell is an NK cell. In some embodiments, the immune cell is a T cell. In some embodiments, the hinge domain comprises a CD8a hinge domain or an Ig4 SH domain. In some embodiments, the transmembrane domain comprises a CD8a transmembrane domain, a CD28 transmembrane domain, and/or a CD3 transmembrane domain. In some embodiments, the signaling domain comprises an OX40 signaling domain, a 4-1BB signaling domain, a CD28 signaling domain, an NKp80 signaling domain, a CD16 IC signaling domain, a CD3-zeta or ITAM CD3ζ signaling domain, and/or an mIL-15 signaling domain. In some embodiments, the signaling domain comprises a 2A cleavage domain. In some embodiments, the signaling domain of mIL-15 is separated from the rest or other portion of the CD19-directed chimeric receptor by a 2A cleavage domain. In some embodiments, such immune cells are administered to a subject having a cancer for the purpose of treating, inhibiting, or preventing the progression of the cancer.

[00259] Also provided herein is an engineered NK or T cell that expresses a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH domain comprises a VH domain derived from humanization of the VH domain amino acid sequence set forth in SEQ ID NO: 33, the VL domain comprises a VL domain derived from humanization of the VL domain amino acid sequence set forth in SEQ ID NO: 32, a hinge and/or transmembrane domain, an intracellular signaling domain, wherein the intracellular signaling domain comprises an OX40 subdomain, and wherein the cell also expresses membrane-bound interleukin-15 (mbIL15).

[00260] Provided herein is a polynucleotide encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH domain comprises a VH domain derived from humanization of the VH domain amino acid sequence set forth in SEQ ID NO: 33, the VL domain comprises a VL domain derived from humanization of the VL domain amino acid sequence set forth in SEQ ID NO: 32; a hinge and/or transmembrane domain,

[00261] an intracellular signaling domain, wherein the intracellular signaling domain comprises an OX40 subdomain, and wherein the polynucleotide further encodes membrane-associated interleukin-15 (mbIL15).

[00262] Provided herein is a polynucleotide encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises an scFv; a hinge, wherein the hinge is a CD8alpha hinge; a transmembrane domain; and an intracellular signaling domain, wherein the intracellular signaling domain comprises a CD3zeta ITAM. In some embodiments, the transmembrane domain comprises a CD8alpha transmembrane domain, an NKG2D transmembrane domain, and/or a CD28 transmembrane domain. In some embodiments, the intracellular signaling domain comprises a CD28 signaling domain, a 4-1BB signaling domain, and/or an OX40 domain. In some embodiments, the intracellular signaling domain can also comprise a domain selected from ICOS, CD70, CD161, CD40L, CD44, and combinations thereof.

[00263] Provided herein is a polynucleotide encoding a CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable region of an scFv or a light chain variable region of an scFv; a hinge, wherein the hinge is a CD8alpha hinge; a transmembrane domain, wherein the transmembrane domain comprises a CD8alpha transmembrane domain; and an intracellular signaling domain, wherein the intracellular signaling domain comprises a CD3zeta ITAM. In some embodiments, the polynucleotide further encodes a truncated epidermal growth factor receptor (EGFRt). In some embodiments, the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). This document provides an engineered NK or T cell that expresses such a CD19-directed chimeric antigen, as well as methods for treating cancer by administering such an NK cell or T cell. Also provided is the use of such polynucleotides in the treatment of cancer, such as in the manufacture of a drug for the treatment of cancer.

[00264] Provided herein is a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH domain comprises a VH domain selected from SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123, the VL domain comprises a VL domain selected from SEQ ID NO: 117, SEQ ID NO: 118, and SEQ ID NO: 119; a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes membrane-associated interleukin-15 (mbIL15). In some embodiments, the intracellular signaling domain comprises an OX40 subdomain, a CD28 subdomain, an iCOS subdomain, a CD28-41BB subdomain, a CD27 subdomain, a CD44 subdomain, or a combination thereof. In some embodiments, the chimeric antigen receptor comprises a hinge and a transmembrane domain, wherein the hinge is a CD8alpha hinge, wherein the transmembrane domain is either a CD8alpha transmembrane domain or an NKG2D transmembrane domain. In some embodiments, the intracellular signaling domain comprises a CD3zeta domain.

[00265] Provided herein is a polynucleotide encoding a humanized chimeric antigen receptor (CAR), wherein the CAR comprises a single chain antibody or single chain antibody fragment that comprises a humanized CD19 binding domain, a transmembrane domain, a primary intracellular signaling domain comprising a native intracellular signaling domain of CD3-zeta or a functional fragment thereof, and a costimulatory domain comprising a native intracellular signaling domain of a protein selected from the group consisting of OX40, CD27, CD28, ICOS, and 4-1BB, or a functional fragment thereof, wherein said CD19 binding domain comprises a light chain complementarity determining region 1 (CDR1 LC) of SEQ ID NO: 124, 127, or 130, a light chain complementarity determining region 2 (CDR2 LC) of SEQ ID NO: 125, 128 or 131, and a light chain complementarity determining region 3 (CDR3 LC) of SEQ ID NO: 126, 129, or 132, and a heavy chain complementarity determining region 1 (CDR1 HC) of SEQ ID NO: 133, 136, 139, or 142, a heavy chain complementarity determining region 2 (CDR2 HC) of SEQ ID NO: 134, 137, 140, or 143, and a heavy chain complementarity determining region 3 (CDR3 HC) of SEQ ID NO: 135, 138, 141, or 144. In several embodiments, the polynucleotide further comprises a region encoding membrane-associated interleukin 15 (mbIL15).

[00266] Provided herein is a polynucleotide encoding a humanized CD19-directed chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a light chain variable domain (VL) of SEQ ID NO: 117, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 161, SEQ ID NO: 167, SEQ ID NO: 173, SEQ ID NO: 179, SEQ ID NO: 185, SEQ ID NO: 191, SEQ ID NO: 197, or SEQ ID NO: 203.

[00267] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a light chain variable domain (VL) of SEQ ID NO: 118, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 163, SEQ ID NO: 169, SEQ ID NO: 175, SEQ ID NO: 181, SEQ ID NO: 187, SEQ ID NO: 193, SEQ ID NO: 199, or SEQ ID NO: 205.

[00268] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a light chain variable domain (VL) of SEQ ID NO: 119, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 165, SEQ ID NO: 171, SEQ ID NO: 177, SEQ ID NO: 183, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 201, or SEQ ID NO: 207.

[00269] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 120, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 185, SEQ ID NO: 187, or SEQ ID NO: 189.

[00270] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 121, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171, SEQ ID NO: 191, SEQ ID NO: 193, or SEQ ID NO: 195.

[00271] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 122, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 173, SEQ ID NO: 175, SEQ ID NO: 177, SEQ ID NO: 197, SEQ ID NO: 199, or SEQ ID NO: 201.

[00272] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety, wherein the CD19 binding moiety comprises a humanized scFv sequence comprising a heavy chain variable domain (VH) of SEQ ID NO: 123, a hinge and/or transmembrane domain, an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15). In several embodiments, the polynucleotide encodes a humanized chimeric antigen receptor of SEQ ID NO: 179, SEQ ID NO: 181, SEQ ID NO: 183, SEQ ID NO: 203, SEQ ID NO: 205, or SEQ ID NO: 207.

[00273] In several embodiments, the provided polynucleotides do not encode sequences of SEQ ID NO: 112, 113, 114, or 116.

[00274] A polynucleotide encoding a humanized CD19-directed chimeric antigen receptor is provided, wherein the chimeric antigen receptor comprises an extracellular CD19 binding moiety; a hinge and/or transmembrane domain; an intracellular signaling domain, and wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15), and wherein the polynucleotide is selected from the group consisting of polynucleotides having at least 95% sequence identity with SEQ ID NO: 184, SEQ ID NO: 186, SEQ ID NO: 192, or SEQ ID NO: 200. In several embodiments, the polynucleotide has a sequence of SEQ ID NO: 184, SEQ ID NO: 186, SEQ ID NO: 192, or SEQ ID NO: 200. In several embodiments, engineered NK or T cells that express such a humanized CD19-directed chimeric antigen receptor are provided. A method of treating cancer in a subject is also provided, comprising administering to a subject having a cancer such engineered NK and/or T cells. The use of such polynucleotides in cancer treatment is also envisaged, such as in the production of a drug for the treatment of cancer.

EXAMPLES

[00275] The materials and methods disclosed herein are non-limiting examples that are used in accordance with certain embodiments disclosed herein.

[00276] According to several embodiments, NK cells are isolated from peripheral blood mononuclear cells and expanded using a feeder cell line. As discussed in more detail below, in several embodiments, the feeder cells are engineered to express certain stimulatory molecules (e.g., interleukins, CD3, 4-1BBL, etc.) to promote the expansion and activation of immune cells. Engineered feeder cells are disclosed, for example, in International Patent Application PCT/SG2018/050138, which is incorporated herein by reference in its entirety. In several embodiments, stimulatory molecules, such as interleukin 12, 18, and/or 21, are separately added to the co-culture medium, such as at specific times and in specific amounts, to promote enhanced expansion of the desired immune cell subpopulation(s).

[00277] NK cells isolated from PBMCs were co-cultured with K562 cells expressing membrane-bound IL15 and 4-1BBL, with IL2 supplemented in the medium. For one set of engineered NK cells, they were expanded in medium supplemented (on day 0) with a combination of soluble IL12 and soluble IL18. The medium was refreshed with additional soluble IL12 and soluble IL18 on day 4. Additional details of embodiments of such a culture methodology are disclosed in U.S. Provisional Patent Application No. 62/881,311, filed July 31, 2019, and incorporated herein by reference in its entirety. Viral transduction with the CD19-targeted chimeric receptor construct was performed on day 7. The resulting engineered NK cells were assessed after 14 or more days of total culture time.

Example 1

[00278] Figure 5 is a schematic of an experimental model for evaluating the antitumor efficacy of engineered NK cells generated according to the methods disclosed herein. NOD-scid IL2Rgamma ^null mice were injected intravenously on day 0 with 2 x 10 ⁵ Nalm6 cells (a B-lymphocyte progenitor leukemia cell line). On day 4, one group of mice was injected with 2.7 x 10 ⁷ NK cells expressing the NK19 CAR (see Figure 3A, although it should be understood that other CD19-directed chimeric receptors can be used), while another group was injected with NK cells that had been expanded using soluble IL12/IL18 (referred to as NK19-IL12/18 cells). Leukemic tumor burden was assessed by fluorescence imaging performed on days 3, 7, 11, 18, and 25. Figure 6A shows the imaging results on days 3, 7, 11, and 18. As can be seen, mice receiving either NK19 or NK19 IL12/18 demonstrated significantly lower tumor burden than mice receiving either untransduced NK cells or PBS as a control. Figure 6B shows a summary of the imaging data as a line graph (higher flux values (photons/second) indicate greater fluorescence signal and greater tumor burden). Both NK19 and NK19 IL12/18 groups demonstrated lower tumor burden as early as day 7 after injection of Nalm6 leukemic cells. This difference is even more pronounced at day 11, when the PBS and NT NK cell groups showed large tumor growth. Even at day 18, when the tumor burden in the PBS and NT NK groups was extensive, the NK19 and NK19 IL12/18 groups showed much lower tumor burden. Surprisingly, the NK19 IL12/18 groups showed lower tumor burden than the NK19 construct groups. This is surprising not only because NK19 cells are very effective at preventing leukemia cell growth, so that this effect would be further enhanced, but also because the previous cell expansion technique affected not only the cell numbers themselves, but also the activity level of these expanded cells.

Example 2

[00279] Experiments were undertaken to determine whether a given stimulatory domain (also referred to as costimulatory domains, given that many constructs use multiple "signaling" domains in a tandem, triplet, or other multiplexed manner) is utilized in the expression (as well as activity) affected by the CAR. NK cells were generated by transduction with viruses encoding the various CARs depicted in Figures 3A-3C (although several embodiments use other constructs). As a non-limiting example, NK cells were cells that were generated by transduction with a bicistronic virus encoding an anti-CD19 scFv, an OX40 intracellular costimulatory domain, a CD3ζ signaling domain, and membrane-associated IL-15 (see NK19, Figure 3A), which supports long-term cell survival and proliferation. Other CAR constructs tested, NK19-1, 2, 3, 4, 5, 8, 9, 10, 11, 12, and NK19-13, were transduced into NK cells in a similar manner. It should be appreciated that for those constructs that utilize the Flag domain to determine expression, similar constructs that do not include Flag (or another tag domain) are provided in several embodiments.

[00280] Figure 7 summarizes the expression data for NK 19-1 through NK5 and NK8 through NK19-13 constructs as evidenced by detection of CD19Flag. The data are presented as the percentage of NK cells expressing CD19-Flag relative to the total number of NK cells present. The data were collected 4 days after transduction with the respective CAR NK19-"X" encoding virus. As evidenced by the expression data, all constructs were expressed in at least 55% of the NK cells. In fact, for eight of the eleven constructs for which data were collected, expression was detected in approximately 75% or more of the total NK cells, with several constructs expressing with greater than 80% efficiency. These expression data indicate that, in accordance with several embodiments, selection of specific stimulatory domains can provide for more efficient expression of a CAR by an NK cell. In some embodiments, this is advantageous because a greater proportion of the NK cell-based formulation is clinically applicable (e.g., fewer administered NK cells are required to obtain a clinically significant dose of engineered NK cells).

[00281] As discussed herein, chimeric antigen receptors that target CD19 (or other tumor markers) can utilize a variety of costimulatory domains. Figure 8A shows data relating to the expression of CARs that target CD19 using a variety of costimulatory domains. As a non-limiting example, costimulatory domains can include, but are not limited to, OX40, CD28, iCOS, CD28/41BB, CS27, and CD44). Figure 8A shows mean fluorescence intensity data representing the expression of these CAR constructs by NK cells. As evidenced by the low MFI observed for the GFP control, these data indicate that the constructs are (a) expressed by NK cells and (b) relatively stably expressed by NK cells over a 4-week period following transduction. Figure 8B shows the expression efficiency of CARs with the indicated costimulatory domains. Although there is some variability in expression efficiency, ranging from approximately 60% to approximately 80% efficient, each of the CARs with the indicated costimulatory domains was expressed well and was expressed relatively stably for at least 4 weeks.

[00282] Data regarding the cytotoxic efficacy of NK cells expressing CD19-directed CARs using different costimulatory domains are shown in Figures 9A-9F. Cultured Nalm6 or Raji cells were exposed to engineered NK cells expressing the indicated NK19 constructs and co-cultured for the indicated number of days (X-axis represents days) of exposure to NK cells expressing NK19-X. Figure 9A shows the cytotoxic effects of the constructs on Nalm6 cells 7 days after transduction of NK cells from the first donor with the construct. The ratio of effector to target cells for this experiment was 1:1. As shown in the graphs, NK19-10, NK19-8, NK-1911, NK19-5, and NK19-12 each allowed for an increase in detectable Nalm6 cells on par with that provided by NK cells expressing only GFP as a control. However, NK19-3, NK19-9, NK19-4, NK19-13, NK19-1, and NK19-2 each showed a significantly smaller increase in Nalm6 cells (e.g., greater cytotoxicity). Therefore, in several embodiments, such constructs are expressed in NK cells and used to treat B cell leukemia (or other tumor types). In several embodiments, one or more stimulatory domains from one of the constructs are inserted into another construct with a different stimulatory domain, which advantageously results in synergistic signaling and further enhanced cytotoxicity. As a non-limiting example, the NK19-1 construct with the OX40 stimulatory domain is, in some embodiments, further engineered to express the CD44 stimulatory domain in addition to OX40. As a further non-limiting example, the NK19-1 construct with the OX40 stimulatory domain is, in some embodiments, further engineered to express the CD44 stimulatory domain and the CD17 stimulatory domain in addition to OX40.

[00283] Figure 9B shows corresponding cytotoxicity data for engineered NK cells from a second donor against Raji B cell leukemia cells 7 days after transduction. The ratio of effector cells to target cells in this case was also 1:1. As shown, similar to the activity of the engineered constructs against Nalm6 cells, some constructs allowed an increase in the number of Raji cells. However, NK19-13, NK 19-4, NK19-2, NK19-3, NK19-1, and NK19-9 each significantly prevented the growth of Raji cells, with several constructs resulting in virtually no growth of Raji cells. Therefore, in several embodiments, such constructs are expressed in NK cells and used to treat B cell leukemia (or other tumor types).

[00284] Figure 9C shows data for NK cells from donor 1 against Nalm6 cells 14 days after transduction. The ratio of effector cells to target cells was 1:1. As shown, even two weeks after transduction, NK cells expressing NK19-13, NK19-4, NK19-3, NK19-2, NK19-1, and NK19-9 prevented the growth of virtually all Nalm6 cells, demonstrating their high level of cytotoxic activity against tumor cells. Accordingly, in several embodiments, such constructs are expressed in NK cells and used to treat B cell leukemia (or other tumor types). As discussed above, in several embodiments, a construct is provided that utilizes a combination of two, three, or more stimulatory domains, resulting in synergistic stimulation of NK cells and enhanced cytotoxicity.

[00285] Figure 9D shows data for NK cells from donor 2 against Raji cells 14 days after transduction. The ratio of effector cells to target cells was 1:2. As shown, NK cells expressing only GFP allowed Raji cells to grow substantially similar to untreated Raji cells. In contrast, NK cells expressing each of NK19-3, NK19-1, NK19-4, NK19-13, NK19-2, and NK19-9 significantly inhibited Raji cell growth, with several constructs allowing little or no growth of Raji cells. Accordingly, in several embodiments, such constructs are expressed in NK cells and used to treat B cell leukemia (or other tumor types). As discussed above, in several embodiments, a construct is provided that utilizes a combination of two, three, or more stimulatory domains, resulting in synergistic stimulation of NK cells and increased cytotoxicity.

[00286] Figure 9E shows summary cytotoxicity data against Nalm6 cells at an E:T ratio of 1:1 and 1:2 at 7 days post-transduction. At an E:T ratio of 1:2, all but one NK19 construct demonstrated equivalent or greater cytotoxicity than NK cells expressing GFP alone. In fact, even at a ratio of 1:2, six constructs achieved cytotoxicity of ~50% or greater. When tested at an E:T of 1:1, seven constructs achieved cytotoxicity of ~50% or greater, but five constructs (NK19-13, NK19-2, NK19-9, NK19-3, and NK19-1) demonstrated cytotoxicity greater than 70%. Figure 9F shows corresponding data for Raji cells. All tested constructs demonstrated enhanced cytotoxicity compared to GFP-expressing NK cells at both 1:2 and 1:1 E:T ratios. At 1:2 E:T, four constructs exceeded 40% cytotoxicity, while at 1:1, seven constructs exceeded this killing rate. Moreover, at 1:1 E:T, three constructs demonstrated 60% or greater cytotoxicity, with the best performing construct achieving nearly 90% cytotoxicity. Taken together, these data demonstrated that a variety of CD19-targeted CAR constructs can not only be expressed, but also stably expressed and effectively induce cytotoxicity against multiple malignant cell types, in some cases exceeding 80% killing rate. In further embodiments, CD19-targeting constructs are provided that utilize combinations of two, three, or more stimulatory domains, resulting in further enhancement of the cytotoxicity of NK cells expressing them. In some embodiments, CD19-targeting constructs can synergistically interact with NK cells expressing receptors directed against other tumor markers, such as NKG2D ligands (such chimeric receptor-bearing NK cells are described in PCT/US2018/024650, which is incorporated herein by reference in its entirety). For example, a chimeric receptor comprising a binding domain that binds NKG2D ligands, an OX40 stimulatory domain, and a CD3-zeta signaling domain can be used in combination with any of the CD19-targeting constructs disclosed herein. In some embodiments, such a chimeric receptor is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 145. In some embodiments, the sequence of such a chimeric receptor is at least 90% identical to the amino acid sequence of SEQ ID NO: 146.

[00287] Additional experiments were performed to evaluate the cytotoxicity of the selected CD19-targeted CAR constructs. NK cells isolated from three different donors (two donors for the 14-day experiment) were transduced with vectors encoding the indicated constructs, NK19-1 (OX40 costimulatory domain); NK19-2 (CD28 costimulatory domain); NK19-3 (ICOS costimulatory domain); NK19-4 (CD28-41BB costimulatory domain); NK19-9 (CD27 costimulatory domain), and NK19-13 (CD44 costimulatory domain). For seven (n = 3) or 14 (n = 2) days post-transduction, these engineered NK cells were co-cultured with Nalm6 or Raji cells at an E:T ratio of 1:1. The results are shown in Figure 10A (expressed as the percentage of increased cytotoxicity compared to GFP-expressing NK cells). Consistent with the data shown in Figure 9, each of the constructs tested demonstrated enhanced cytotoxic effects on Nalm6 cells, ranging from a mean increase of 40% for NK19-4 to an overall mean increase of approximately 50% for the other 5 constructs. Figure 10B shows the corresponding data for Raji cells. Again, each construct was superior to GFP-only expressing NK cells, with mean increases in cytotoxicity compared to GFP-expressing NK cells ranging from approximately 40% to approximately 50%. Using cells from two donors 14 days post-transduction, NK19-1, NK19-9, and NK19-13 were tested on Nalm6 and Raji cells. Figure 10C shows the calculated enhancement in cytotoxicity of these constructs compared to GFP-expressing NK cells, where an average enhancement of nearly 80% was observed for NK cells expressing NK19-9, approximately 75% for NK19-1 expressing cells, and greater than 60% for NK19-13 expressing cells. Similar results were observed for Raji cells, with NK19-13 expressing cells showing greater than 20% improvement in cytotoxicity, NK19-1 expressing cells showing nearly 60% enhancement, and NK19-9 expressing cells showing nearly 70% greater cytotoxic activity against Raji cells. These results further support the embodiments disclosed herein that provide engineered NK cells expressing CD19-targeted CARs and methods for using them in the treatment of cancer, in that immunotherapy results in enhanced cytotoxicity against target tumor cells.

[00288] Figures 11A-11E provide data regarding the cytokine release profiles of NK cells in Nalm6 cells, related to the mechanism by which NK cells control tumor and virus-infected cells - through the release of cytotoxic granules and proinflammatory cytokines. Figure 11A shows that, compared to control or even GFP-expressing NK cells, NK cells expressing NK19-1, NK19-9, or NK19-13 all express higher levels of the serine protease, granzyme B, which is present in granules released by NK cells. NK cells expressing these CD19-directed CARs released approximately 4-fold more granzyme B than control GFP-expressing NK cells. Figure 11B shows data regarding the increased release of perforin by the engineered NK cells. Interestingly, perforin levels were not significantly higher than those produced by GFP-expressing NK cells (although perforin concentrations were elevated compared to controls). Perforins act in concert with granzyme B (and other granzymes), with perforins functioning to form pores in the cell membrane, allowing granzymes to cross the membrane and then exert their protease effects on intracellular protein targets. These data raise the possibility that perforin release, although roughly similar or reduced with certain constructs, is actually more efficient at pore formation, allowing for similar levels of pore formation. Alternatively, if perforins released by NK cells expressing NK19 are no more efficient at forming pores, this is compensated for by increasing granzyme B (and/or other granzymes), thus enhancing cytotoxicity is still achieved.

[00289] Figure 11C shows that, compared to control or even GFP-expressing NK cells, all NK cells expressing NK19-1, NK19-9, or NK19-13 release higher concentrations of the inflammatory cytokine TNF-alpha. Figure 11D shows similar data for GM-CSF release by NK cells expressing NK19, and Figure 11E shows similar data for interferon-gamma release. Similarly, when tested in Raji cells, similar release patterns are obtained as shown in Figures 12A-12E. These data indicate that NK19-expressing cells exert their cytotoxic effects, at least in part, by increasing the release of inflammatory cytokines and/or cytotoxic granules. As mentioned above, in several embodiments, engineered CARs are designed to contain combinations of two, three, or more costimulatory domains to synergistically enhance cytokine/granule release and cytotoxicity against target cancer types.

[00290] To further confirm the enhanced effects of NK19 constructs on tumor progression, NSG mice were injected intravenously with 1 x 10 ⁵ Nalm6 cells (expressing a fluorescent reporter) on day 0. On day 3, mice were injected with either control PBS, untransduced NK cells ("NTNK", 10 M), NK19-2-expressing NK cells (10 million cells), NK19-9-expressing cells (10 million), NK19-1-expressing cells (10 million), or NK19-1-expressing cells (30 million). Fluorescence imaging for Nalm6 cells was performed on days 3, 8, 11, 18, and 25 (imaging data not shown). The data are shown in Figure 13. As shown, injection of NK cells expressing either NK19 variant resulted in reduced progression of Nalm6 outgrowth. NK cells expressing NK19-2 showed little growth in Nalm6 at day 8, with an increase in Nalm6 outgrowth by day 11 and a significant increase by day 18 (though less than with NTNK cells). Neither NK19-9 nor NK19-1 expressing NK cells (at any dose) showed detectable tumor burden at day 8 by imaging. Mice receiving NK19-9 expressing NK cells did show some increase in Nalm6 cell outgrowth by day 11 with further progression by day 18. Mice receiving any dose of NK19-1 expressing cells showed no growth in Nalm6 cells at day 11. At day 18, some tumor growth was observed in mice receiving 10 million NK19-9-expressing cells. However, at the 30 million cell dose, only modest growth of Nalm6 cells was observed in these mice receiving NK cells expressing NK19-9.

[00291] Figure 14A is a line graph of the bioluminescence intensity detected in mice (e.g., the fluorescent signal shown in Figure 13, note that Figure 13 does not show the imaging data for day 25). Consistent with the images in Figure 13, the line graph in Figure 14A shows an increase in Nalm6 cells (as represented by an increase in BLI) for all groups at day 25, with significant numbers of cells detected in the PBS and NTNK groups and slightly fewer in the NK19-2, NK19-1 (10 M), and NK19-9 groups. The NK19-1 (30 million) group showed the smallest increase, demonstrating the ability of the construct to reduce the level of Nalm6 progression through cytotoxic effects on Nalm6 cells. While each construct allows some Nalm6 growth over time, even if moderate, the NK19 constructs tested delay the onset of this growth, as evidenced by the flat line until day 11 for the NK19 curves. To better demonstrate this aspect, the data are plotted on the logarithmic scale y-axis in Figure 14B, allowing for the curves to be separated. As shown, the NTNK and PBS curves show an increasing trend after day 3, indicating an almost immediate increase in Nalm6. In contrast, the NK19-9, NK19-2 and both NK19-1 curves either decline or show only a slight upward trend until day 7. At day 11, consistent with the images in Figure 13, Nalm6 cell growth is detected in the NK19-9, NK19-2 and NK19-1 groups (10 M). In contrast, the NK19-1 group (30 million) is still approximately at baseline, reflecting the absence of any significant growth of Nalm6 cells. Cell growth reverses to increase in the NK19-9 group (30 million group) at day 18. Although there is an increase in tumor cells, even with administration of NK19 constructs, the delay in the onset of growth may be advantageous in several embodiments. For example, it allows the patient to be re-administered with another dose of engineered NK cells expressing a CD19-targeted construct. In several embodiments, the subsequent dose (like the initial dose) may optionally contain NK cells that have been altered to reduce allogeneity, such as by gene editing. Thus, in several embodiments, two, three, four or more doses of NK cells expressing a CD19-targeted CAR are administered. This tumor cell growth delay provides the opportunity to sequentially (or simultaneously) administer doses of NK cells expressing a chimeric construct targeting a different tumor marker and/or administer some other type of anti-cancer therapy (e.g., immune checkpoint inhibitor, antibody therapy, chemotherapy, etc.). Figure 14C presents data relating to the expression of CD3 by cells transduced with the various constructs in blood samples obtained from mice treated as indicated on the x-axis. CD3 is a T cell marker. As indicated, but for T cells engineered to express the NK19-1 construct and a mixed population of NK cells and T cells, CD3 expression was essentially negligible. Figure 14D presents data relating to the expression of CD56, which is a human NK cell marker. Although there is a small amount of background staining, staining of blood samples from mice treated with NK cells expressing the indicated constructs is relatively low (as would be expected given that (i) the blood is a mouse blood sample and mouse cells will make up the majority of the total, and (ii) CD56 only detects human NK cells (e.g., injected). The data are notable in this regard in that the group treated with 30 million NK19-1 showed markedly higher CD56 expression than the other groups, and the T cell/NK+ T cell groups expressed CD56 at background levels. Figure 14E shows data related to GFP expression by tumor cells. Blood samples were collected ~3 weeks after the start of the in vivo experiment. As shown (consistent with the images/BLI data), the PBS and NTNK control groups exhibit higher percentages of GFP expression (e.g., a higher percentage of total live blood cells in sample are tumor cells). Each of the engineered constructs shows significantly less GFP expression, based on the results of the engineered constructs controlling/reducing tumor cell growth. Figure 14F shows data similar to Figure 14D, but CD19 expression was measured in all live cells in this mouse blood sample. As with GFP, the PBS and NKNT groups show CD19 expression of approximately 10-12%, meaning that 10-12% of the live cells in the blood sample are tumor cells and the rest are mouse blood cells. As shown in the other experimental group, CD19 expression is at a much lower level, reflecting the fact that the engineered CAR constructs limit tumor cell growth. These data are consistent with the embodiments disclosed herein, wherein engineered NK cells expressing CD19-directed CARs are highly cytotoxic and provide the potential for the treatment of cancer tumors.

Further evaluation of humanized designs

[00292] As discussed in detail above in several embodiments, several embodiments of the CARs disclosed herein include the use of humanized sequences, such as an extracellular binding moiety. In several embodiments, a humanization approach is applied to one or more aspects of the art. In several embodiments, one or more heavy and/or light chains of an antibody are humanized, which (as discussed above) can provide advantages including, but not limited to, reduced immunogenicity, increased stability, longer-lasting efficacy, increased activity, and the like.

Example 3

[00293] Figure 15 shows a diagram of a series of humanized constructs according to several embodiments disclosed herein. Such constructs are designated by the letter "H" in their identifier. By way of illustration, NK19H-1 is a humanized anti-CD19 CAR that utilizes an scFv consisting of a first light chain and a first heavy chain ("L1H1"), while NK19H-3 utilizes an scFv consisting of a third light chain and a first heavy chain ("L3H1"). Figure 15 also shows data relating to the stability and aggregation of various combinations of heavy and light chains following transient expression and secretion by 293T cells. Shown are data relating to the mean fluorescence intensity detected by flow cytometry when each antibody was heated to 70 ^o C and then cooled back to room temperature ("heated"), compared to the same variant kept on ice ("no heat"). Following heat treatment, the ScFv variants were used in the flow cytometry protocol at different concentrations (0.4, 0.25, and 0.125 μg/mL). A loss in fluorescence intensity indicates that the ScFv either lost structural integrity or aggregated during the heating process. ScFvs with the best thermal stability, as indicated by comparable MFI under both conditions, are preferred for further development according to some embodiments.

[00294] Following assessment of construct stability, the selected anti-CD19 CAR constructs were further assessed. Figure 16 shows a summary of the expression data for the designated constructs by NK cells from 3 donors (#140, #9, and #20). As shown in the figure, expression was assessed by detecting CD19Flag. Data are presented as the percentage of NK cells expressing CD19-Flag relative to the total number of NK cells present. Data were collected 4 days after transduction with the corresponding virus encoding the NK19H-"X" CAR. As evidenced by the expression data, each of the selected constructs was expressed by NK cells to varying degrees. Expression levels ranged from being expressed in approximately 20% of all NK cells with NK19H-2, NK19H-11, and NK19H-12. Most of the other constructs were successfully expressed in ~40%-60% of NK cells, which is on par with the expression of the non-humanized NK19-1 construct. The NK19H-5 construct was expressed in greater than 80% of NK cells. Although some constructs may be expressed more efficiently, constructs with lower expression levels were still evaluated because, in several embodiments, such constructs still exhibit significant cytotoxicity to target cells. However, in several embodiments, constructs with higher expression efficiencies may be preferred, for example, because a greater proportion of the NK cell preparation is clinically applicable (e.g., fewer administered NK cells are required to obtain a clinically relevant dose of the engineered NK cells).

[00295] Figures 17A-17E depict such cytotoxicity data. Figure 17A depicts the cytotoxicity of NK cells from donor 20 transduced with the designated constructs directed against the CD19-positive leukemia cell line Nalm6 (at an E:T of 1:1; 20,000 cells/well). The data show that, despite varying expression levels, most of the NK19H constructs were able to exhibit cytotoxic effects on Nalm6 target cells. NK19H-11 exhibited the least potency, allowing Nalm6 cells to grow at levels slightly below control levels. In contrast, NK19H-1 and NK19H-3 exhibited cytotoxicity on par with the non-humanized NK19-1 construct, allowing some cell growth at later time points in co-culture. Notably, NK19H-4 and NK19H-5 exhibited significant cytotoxicity, allowing only very limited growth of Nalm6 throughout the experiment.

[00296] Figure 17B shows NK cells from donor 20 tested against the CD19-positive Burkitt lymphoma cell line Raji. As with Nalm6 cells, the different humanized CD19 constructs exhibited varying cytotoxicity against target cells. Similar to the data in Figure 17A, the NK19H-11 construct exhibited limited cytotoxicity, but all other constructs exhibited promising cytotoxicity against target cells, with NK19H-1 performing on par with the non-humanized NK19-1 construct and each of NK19H-3, 19H-4, and 19H-5 induced higher levels of cytotoxicity, restricting the growth of Raji cells to later stages of co-culture (with NK19H-5 allowing only very limited growth of Raji cells). Figure 17C shows the corresponding data for Nalm6 cells from donor 140. Here, a similar pattern of efficacy was observed, with the NK19H-11 construct allowing Nalm6 cells to grow to the negative control. However, NK19H-1, 19H-3, and 19H-4 each induced at least as much cytotoxicity as non-humanized NK19-1. Again, NK19H-5 showed significant cytotoxicity, limiting Nalm6 cell growth throughout the experiment. Figure 17D shows data from donor 9 against Raji cells. Only the NK19H-11 construct allowed any significant growth of Raji cells. In contrast, NK cells from this donor expressed non-humanized NK19-1 or any of the humanized constructs NK19H-1, 19H-3, 19H-4, or 19H-5, suppressing any growth of Raji cells by inducing cytotoxic effects. Figure 17E shows data for NK cells from donor 9 against Nalm6 cells. In this experiment, the cytotoxicity of the three humanized constructs (NK19H-11, 19H-1, and 19H-3) was limited. There is some variability among donors, as some of these constructs induced cytotoxicity when expressed by NK cells from other donors. Constructs NK19H-4 and 19H-5 exhibited significant cytotoxic effects, limiting Nalm6 cell growth to virtually zero over the course of the experiment. Taken together, these data complement the expression data and show that, in several embodiments, humanized CAR constructs targeting CD19 are effective in killing tumor cells even though they have different expression efficiencies. Additionally, in some embodiments, expression efficiency does not correlate with cytotoxicity, and even constructs with limited expression efficiency can exhibit significant cytotoxicity.

Example 4

[00297] Additional experiments similar to those discussed above were performed using NK cells from additional donors. Figure 18 shows the construct expression efficiency for three donors (#945, 137, and 138) for the indicated constructs. As in the previous experiment, the data provided represent the number of CD19-Flag positive cells out of the total number of NK cells evaluated. The data for these donors show a higher overall expression efficiency for all but one of the humanized constructs. Only NK19H-3 was expressed to a lesser extent than the non-humanized NK19-1 construct. For NK cells from these donors, expression of the humanized constructs was detected in approximately 70-80% of the NK cells. As noted above, in addition to assessing expression, cytotoxicity against CD19-expressing target cells was assessed. Figure 19A shows data for NK cells from a donor (#137) expressing the indicated constructs and co-cultured with Raji cells. In this experiment, engineered NK cells expressing the indicated constructs were exposed to tumor cells at two time points, 7 days apart, and an additional bolus dose of tumor cells was added 14 days after transduction. The arrow indicates the second injection of tumor cells. As shown, untreated Raji cells expanded throughout the experiment. NK cells expressing GFP or non-humanized NK19-1 induced some cytotoxicity, as evidenced by decreased Raji cell growth compared to controls. NK cells expressing NKH19-3 (the humanized construct with the lowest expression efficiency) were also able to reduce Raji cell growth. Each of the other humanized NK constructs exhibited the ability to reduce Raji cell growth compared to controls even 14 days post-transduction, demonstrating the enhanced persistence of the engineered NK cells disclosed herein. Figure 19B shows corresponding data for donor 137 NK cells against Nalm6 cells, with engineered NK cells added back at day 7 (day 0 of the experiment) and day 14 post-transduction (~day 7 of the experiment). The cytotoxicity of these constructs was more variable in this particular experiment. However, several humanized constructs directed against CD19 exhibited the ability to provide significant cytotoxicity and reduce Nalm6 cell growth compared to controls. These data suggest that, in accordance with several embodiments, for certain subjects, a more frequent dosing regimen (e.g., every 2 days, every 3 days, every 4 days, every 5 days, etc.) would be preferable. Advantageously, in several embodiments, the engineered NK cells disclosed herein are allogeneic and can be readily used in more frequent dosing regimens. Figure 19C shows corresponding data for donor 138 against Raji cells. In this case, many of the humanized constructs still exhibited significant cytotoxic effects against Raji cells, even when a second dose of Raji cells was administered 14 days after transduction. Six of the 7 humanized constructs demonstrated such performance and outperformed the non-humanized NK19-1 construct. Figure 19D shows corresponding data for donor 138 against Nalm6 cells. Although the additional bolus dose of Nalm6 cells resulted in increased Nalm6 outgrowth in the latter stages of the experiment, nearly all of the humanized constructs performed better than controls. In fact, NK cells carrying the NK19H-5 construct were able to limit Nalm6 outgrowth until the last few days of the experiment (after the second dose). As noted above, these data demonstrate that humanized anti-CD19 CAR constructs can not only be expressed by NK cells, but can also exhibit cytotoxic activity against highly persistent target cells. In some embodiments, this allows for multiple dosing to be spaced over longer periods of time, which may be advantageous for treating cancers, while limiting potential immunogenicity (at least in part due to humanization and/or reduced frequency of administration).

Example 5

[00298] Additional data regarding the various humanized constructs were collected from additional donors. Figures 20A-20B show the expression data of the various anti-CD19 CAR constructs in NK cells from two additional donors. Figure 20A shows the average fluorescence intensity of NK cells 10 days after transduction. As indicated, in accordance with several embodiments disclosed herein, each of the humanized anti-CD19 CAR constructs exhibited increased overall expression compared to the non-humanized anti-CD19 CAR construct. Figure 20B shows the expression data of CD19-Flag positive NK cells as a percentage of the total NK cells analyzed. As shown, each of the humanized anti-CD19 CAR constructs was expressed more efficiently than the non-humanized construct (which was already expressed in nearly 80% of the NK cells). The humanized CARs were expressed in approximately 85% to 95% of NK cells, depending on the construct. Figure 21A shows cytotoxicity data for engineered NK cells from donor 703 (one of the two donors in Figure 20) against Raji cells co-cultured with NK cells at day 7 post-transduction and again at day 14 post-transduction. As shown in the figure, Raji cells and NK cells expressing only GFP grew robustly until day 7 and again until day 14. The non-humanized anti-CD19 CAR construct, NK19-1, exhibited the ability to suppress Raji cell growth until day 7 and allowed relatively limited growth until day 14. Each of the humanized anti-CD19 CAR constructs additionally suppressed Raji cell growth, with several constructs nearly completely suppressing Raji cell growth. Figure 21B shows the corresponding cytotoxicity data against Raji cells for NK cells isolated from donor 877. These data demonstrate a trend similar to that of the previous donor. The two control groups allowed significant expansion of Raji cells, while each of the humanized anti-CD19 CAR constructs demonstrated significant cytotoxic effects.

[00299] Figures 22A-22E show cytokine release profiles from Raji cells co-cultured with NK cells expressing the indicated constructs. Figure 22A shows IFNg release by NK cells co-cultured with Raji cells. Each of the indicated humanized constructs resulted in increased IFNg release compared to the non-humanized NK19-1 construct. Similarly, as shown in Figure 22B, the humanized anti-CD19 constructs provided NK cells with the ability to release greater amounts of GM-CSF compared to control NK cells. Figure 22C shows that the humanized anti-CD19 CAR constructs induce increased TNF release from NK cells. Perforin levels were not significantly different for all constructs tested, as shown in Figure 22D. Similarly, granzyme levels were relatively constant across all constructs. Taken together, these data indicate that, in some embodiments, humanized anti-CD19 CAR constructs expressed on NK cells cause these NK cells to release increased amounts of one or more cytokines that result in cytotoxic effects on target cells.

Example 6

[00300] Although the data presented above demonstrate that humanized anti-CD19 CARs can be expressed by NK cells and exert cytotoxic effects on target tumor cells, additional experiments were performed to determine the survival of the engineered NK cells. NK cells expressing the indicated anti-CD19 CAR constructs were cultured and cell numbers were measured at various time points up to 26 days post-transduction. Figure 23A shows the survival data for NK cells from donor 137. The data for GFP-expressing NK cells show that the cell count at day 7 was higher than the cell count at any other time point, suggesting that GFP does not have additional effects that promote NK cell survival. Similarly, NK cells expressing non-humanized NK19-1 exhibited a decrease in cell count over time. Although each of the humanized constructs exhibited some variability in cell counts, the trend in the data indicates that expression of the humanized anti-CD19 constructs resulted in less NK cell killing over time. A similar trend is shown in the data in Figure 23B, which shows cell viability for NK cells from donor 138. Although the assay timing was altered, Figure 23C shows data that show a similar trend (for donor 703), in that expression of the humanized constructs resulted in longer NK cell survival in culture over time. Figure 23D shows data for NK cells from donor 877, where expression of the humanized anti-CD19 CAR constructs was able to reduce the rate of NK cell killing in culture over time.

[00301] A summary of the expression data for the various anti-CD19 CAR constructs is shown in Figure 24, which depicts the expression efficiency of the constructs at day 11 post-transduction. As expected, NK cells transduced with GFP did not express CD19-Flag, serving as a negative control. Likewise, the non-humanized NK19-1 construct served as a positive control. Each of the humanized constructs evaluated showed increased expression efficiency compared to NK19-1, with efficiencies ranging from approximately 85% to approximately 95% (e.g., 85-95% of all NK cells tested expressed the Flag-tagged CAR construct).

[00302] Figures 25A-25I show raw flow cytometry data for a single donor where CD19-Flag expression (indicative of CAR expression) was measured. Figure 25A shows a GFP control with little or no Flag expression. Figure 25B shows Flag detection using the NK19-1 positive control. Figures 25C-25I show Flag detection results for the designated humanized anti-CD19 CAR constructs, with the percentage of cells expressing the constructs ranging from approximately 80% to approximately 95%. As in the experiments described above, these data confirm that the humanized anti-CD19 CAR constructs can be stably expressed by transduced NK cells.

Example 7

[00303] Similar to the experiments discussed above, Raji cells were exposed to NK cells from donor 103 that were transduced with the various constructs indicated (see Figure 26A). Raji cells were co-cultured on day 7 following NK cell transduction and re-introduced with NK cells on day 14. As expected, Raji cells alone exhibited continued growth, while GFP-transduced NK cells slightly reduced this cell growth. In contrast, the non-humanized NK19-1 construct, as a positive control, reduced the growth of Raji cells until very late in the experiment. Each of the humanized anti-CD19 CAR constructs exhibited enhanced cytotoxicity against Raji cells. Three of these constructs (NK19H-2, H-3, and H-4) allowed little growth of Raji cells late in the experiment, while the other humanized constructs effectively abolished the growth of Raji cells even when rechallenged at day 14. Figure 26B shows the corresponding data for the cytotoxicity of NK cells from donor 103 against Nalm6 cells. As shown in the figure, all of the anti-CD19 CAR constructs (whether humanized or not) demonstrated significant cytotoxicity against Nalm6 cells over the 7 days of co-culture. However, Nalm6 growth increased in all groups after rechallenge at day 14. Although the growth curves were initially somewhat flat, the growth rate of Nlam6 increased thereafter. These data suggest that some embodiments employ a more frequent dosing strategy to prevent target cells from reaching a threshold growth rate. In some embodiments, a higher dose is administered and/or the dose is administered more frequently to prevent growth of target cells under physiological conditions equivalent to "repeated dosing."

[00304] Figure 26C shows data for Raji cells (as in Figure 26A) with NK cells from donor 275. Similar to the constructs in Figure 26A, each of the anti-CD19 CAR constructs exhibited significant cytotoxicity against Raji cells while allowing limited growth of the target cells even upon repeat dosing. Figure 26D shows data for Nalm6 cells (as in Figure 26B) with NK cells from donor 275. The results of this experiment were also similar to the results for donor 103, with effective control of Nalm6 cells for 7 days, but with decreased cytotoxicity upon repeat dosing. These data indicate that dosing strategies, depending on the embodiment, are designed for a particular donor and/or a particular target tumor type. For example, although Nalm6 cells are CD19 positive, they may express less CD19 than, for example, Raji cells, which explains the reduced cytotoxicity of NK cells from donors 103 and 275 against the Nalm6 cell line. The efficacy of transduced NK cells against Raji cells indicates that NK cells are capable of exhibiting cytotoxicity, even demonstrating persistence for nearly two weeks after transduction. Thus, according to several embodiments, the NK cell dose and/or dosing frequency can be tailored to a given patient's tumor type, native NK cell activity, and/or cancer aggressiveness/stage to provide robust and sustained cytotoxicity to target cells and achieve tumor burden control.

[00305] Figures 27A-27B show flow cytometry data characterizing NK cells from two donors (276 in Figure 17A and 877 in Figure 27B) transduced with non-limiting examples of anti-CD19 CAR constructs disclosed herein. Figures 27A and 27B show that NK cells transduced with GFP express little or no CD19-Flag. Each of the other panels demonstrates that NK cells from these donors can express not only the non-humanized positive control NK19-1 construct, but also efficiently express the selected humanized anti-CD19 CAR constructs. This indicates that humanization of the anti-CD19 binding moiety has a limited impact on the expression characteristics.

Figures 28A-28B show the raw NK cell cytotoxicity data of the two donors shown in Figure 27 against Raji cells at an E:T ratio of 1:1. Consistent with the results discussed above, each of the selected humanized anti-CD19 CAR constructs confers significant cytotoxic potential on Raji target cells to the transduced NK cells. Each of the non-limiting examples of the anti-CD19 CAR constructs provided effective control of Raji cell growth (equal to or enhanced relative to the non-humanized NK19-1 construct), with a trend toward reducing Raji cell numbers even up to 70 hours after the start of the experiment. Similarly, Figure 28B shows that transduced NK cells from donor 877 also effectively controlled the growth of Raji cells, equivalent to or enhanced compared to the non-humanized NK19-1 construct. These data are consistent with the data presented for NK cells from other donors discussed above and indicate that, in accordance with several embodiments disclosed herein, transduction of NK cells with humanized anti-CD19 CARs enables the engineered NK cells to exert significant cytotoxic effects on target tumor cells and can effectively control tumor cell growth. In accordance with several embodiments, such engineered anti-CD19 NK cells (allogeneic or autologous) enable sustainable and effective cellular immunotherapy for the treatment of cancer.

Example 8

[00306] Additional experiments were conducted to evaluate the humanized anti-CD19 CAR constructs disclosed herein. Figure 29A shows a schematic of the experimental protocol. Briefly, NSG mice were injected intravenously on day 0 with 1 x ¹⁰⁵ Nalm6 cells (expressing a fluorescent reporter). On day 1, mice were injected with either control PBS, untransduced NK cells ("NTNK", 10 M), 10 million NK cells expressing the non-humanized NK19-1 CAR construct, or 10 million NK cells expressing one of the various humanized anti-CD19 CAR constructs. Blood collection and fluorescence imaging were performed as indicated. The bioluminescence data are presented in Figure 29B. As shown, injection of NK cells expressing any of the anti-CD19 CAR constructs resulted in a reduction in Nalm6 cell growth progression. Each humanized construct tested (NK19-H2, NK19-H5, and NK19H-9) showed a significant delay in Nalm6 cell growth compared to controls. Figure 29C shows a line graph representing bioluminescence measured over 31 days. As shown, each of the humanized constructs tested in this experiment showed a reduction in tumor progression compared to controls and a slight reduction compared to the non-humanized NK19-1 construct. Figure 29D shows a survival curve that reflects the reduction in tumor progression, with mice treated with NK19-1 or any of the selected humanized constructs surviving longer than mice in the control groups. Figure 29E shows data relating to the expression assessment of each of the humanized constructs on NK cells. As shown, each of the humanized constructs was expressed more stably than the non-humanized NK19-1 construct. According to several embodiments disclosed herein, use of a humanized construct results in greater therapeutic efficacy, at least in part due to increased expression (and/or activity) of CAR by NK cells.

[00307] Figures 30A-30F show data relating to the characteristics of cells in the blood of mice treated with the constructs at various time points during the experiment. Figure 30A shows the percentage of human CD56+ cells in the peripheral blood of mice, which represents the survival of the administered NK cells during the first fifteen days of the experiment. Each of the experimental groups demonstrated a higher percentage of cells, with the NK19-H2 construct demonstrating the greatest persistence of the constructs tested. Figure 30B shows similar data based on the detection of expression of the flag tag used in the constructs (although in some embodiments, the tag was not used). These data indicate that the humanized constructs are expressed at higher levels than the non-humanized constructs. Figure 30C shows the detection of GFP-positive tumor cells after 15 days. CAR-expressing NK cells showed near zero GFP expression, reflecting their inhibition of Nalm6 cell growth at day 15. Figures 30D-30F show similar data at day 32. These data demonstrate that NK cells expressing humanized anti-CD19 CAR constructs account for a higher percentage of cells present in the peripheral blood of the mice tested, consistent with the enhanced efficacy of these constructs in controlling tumor growth at later time points. Figure 30F shows that there is little difference in expression between the humanized anti-CD19 CAR constructs at day 32. Figure 32F depicts the enhanced ability of NK cells expressing humanized anti-CD19 CAR constructs to control tumor growth, with lower numbers of GFP-positive cells detected in the humanized construct treatment groups, even when compared to animals receiving the NK19-1 construct. In some embodiments, the increased efficacy is due, at least in part, to increased expression of humanized CD19 CAR constructs.

Example 9

[00308] As discussed above, in some embodiments, the CAR constructs comprise a label for detection. However, in some embodiments, a label is not used. Experiments were conducted to assess the expression of unlabeled humanized CARs. In this experiment, NK19H-NK-2, -5, and -9 were used as non-limiting examples of unlabeled humanized CARs. Figures 31A-31B show expression data (percentage expression in Figure 31A, average fluorescence in Figure 31B) of the construct in NK cells from three different donors, measured weekly for 4 weeks. These data demonstrate that the unlabeled versions of the anti-CD19 CAR constructs provided herein are expressed relatively similarly to each other, but more consistently than the non-humanized constructs. Each of the unlabeled humanized constructs was expressed in at least about 70-80% of the NK cells.

Example 10

[00309] Experiments were conducted to assess the cytotoxicity of NK cells expressing humanized, untagged anti-CD19 CAR constructs. The in vitro re-challenge assay described above was conducted using Raji cells (Figures 32A-32B) or Nalm6 cells (Figures 32C-32D) as target cells. Figure 32A shows the percentage of Raji cells co-cultured with the indicated treatment, measured as a percentage of the number of cells measured in the control group containing only Raji cells, on day 10 of the experiment. As shown, each of the anti-CD19 CAR constructs, whether humanized or not, and whether labeled or not, essentially abolished Raji cell growth through day 10. Figure 32B shows the last time point at which limited Raji cell growth remained in each of the anti-CD19 CAR experimental groups, although the data show a slight trend toward greater growth prevention of Raji cells in the NK19H-NF-2 and NK19H-NF-5 groups. Figures 32C and 32D show the corresponding data using Nalm6 cells. Figure 32C shows that, as with Raji cells, each of the anti-CD19 CAR constructs, whether humanized or not, and whether labeled or not, essentially abolished Nalm6 cell growth through day 10. At the last time point, Nalm6 cell growth was observed in all groups, although the data show that the humanized constructs were more effective in preventing growth.

[00310] Figures 33A-33J show data pertaining to the detected cytokine profile of NK cells expressing the indicated constructs. As discussed above, culture media from each treatment group were analyzed for gamma interferon (33A/33F), GM-CSF (33B/33G), TNF-alpha (33C/33H), perforin (33D/33I), and granzyme B (33E/33J). Figures 33A-33E show data from a re-inoculation of Raji cells, and Figures 33F-33J show data from a re-inoculation of Nalm6 cells. As shown, NK cells expressing untagged humanized anti-CD19 CAR constructs released similar levels of similar amounts of IFN-gamma into the medium, with levels slightly higher than those released by NK cells expressing the non-humanized NK19-1 CAR construct. Likewise, GM-CSF release was fairly uniform among NK cells bearing humanized untagged CARs, with levels slightly approaching those of NK19-1 expressing cells. TNF-alpha release showed a similar pattern, while perforin release was uniform among all NK cells expressing anti-CD19 CARs, with levels lower than those of control cells. Finally, in Raji cells, granzyme B showed a similar degree of release in the case of unlabeled humanized anti-CD19 CARs at a level slightly higher than that of NK19-1 expressing cells. Advantageously, the data collected in the Nalm6 cell experiment showed similar patterns. According to several embodiments, a higher degree of release of cytotoxicity-mediating cytokines results in greater therapeutic efficacy. In some embodiments, a higher degree of cytokine release is due, at least in part, to enhanced expression of unlabeled humanized anti-CD19 CAR constructs by NK cells and/or, at least in part, is due to increased persistence of NK cells expressing unlabeled humanized anti-CD19 CAR constructs. As further support for these points, Figure 34 shows data regarding the persistence of NK cells expressing the indicated unlabeled humanized constructs over four weeks of culture compared to NK cells expressing NK19-1. Although the overall cell numbers appear to be similar among NK cells expressing the humanized constructs, the cell numbers exhibit the same basic pattern as the cytokines, being slightly higher than NK cells expressing non-humanized NK19-1.

Example 11

[00311] Similar to the experiments described above, an in vivo evaluation of the efficacy of the unlabeled, humanized constructs was conducted. Figure 35A shows a schematic of the protocol used. Figure 35B shows the in vivo bioluminescence measurements, which show that the humanized, unlabeled constructs appeared to slow tumor progression to a greater extent than NK cells expressing the NK19-1 construct. Figure 35C summarizes the visualization data as a line graph, where enhanced tumor cell growth inhibition can be clearly seen when NK cells expressing any of the unlabeled humanized anti-CD19 CAR constructs are administered to mice. Figure 35D shows the enhanced expression of the unlabeled humanized constructs by NK cells. Figures 36A-36F present data pertaining to the persistence of NK cells expressing the indicated constructs over the time course of the experiment. Figure 36A shows that NK cells expressing untagged humanized anti-CD19 CAR constructs are more persistent in vivo, accounting for a greater percentage of total CD56+ cells in peripheral blood at day 13. Figure 36B shows that at day 13, NK cells expressing any anti-CD19 CAR inhibit tumor growth better than controls, with NK cells expressing untagged humanized anti-CD19 CAR constructs showing a slight advantage over NK cells expressing NK19-1 in limiting the growth of CD19+ tumor cells. Figure 36C shows data similar to Figure 36B using the number of GFP-positive tumor cells as the comparative criterion. Figure 36D shows persistence data at day 27, where NK cells expressing NK19-NF-2 and -9 are characterized by an increased population size compared to other groups, indicating increased persistence over a longer period of time. Similar to the previous time point, Figures 36E and 36F show that all CAR constructs targeting CD19 are effective in limiting tumor cell growth compared to control constructs. According to several embodiments provided herein, expression of an unlabeled humanized CAR targeting CD19 by NK cells results in such NK cells having increased persistence and cytotoxicity in vivo.

Example 12

[00312] As discussed herein, in several embodiments, engineered NK cells are produced for allogeneic cell therapy. As such, in several embodiments, engineered NK cells intended for administration are produced and then frozen for subsequent use in a subject. Experiments were conducted to determine whether the cryopreservation process followed by thawing would have an undesirable effect on the engineered NK cells, such as by reducing their viability, persistence, or cytotoxicity. Figure 37A shows a schematic of the experimental protocol used, as well as the experimental groups and other conditions used. For cells designated "IL12/IL18," the cells were expanded in the presence of soluble IL12 and/or IL18 as described in U.S. Provisional Patent Application No. 62/881,311, filed July 31, 2019, and U.S. Patent Application No. 62/932,342, filed November 7, 2019, each of which is incorporated by reference in its entirety. Figures 37B and 37C show in vivo bioluminescence images for the indicated experimental groups. Figures 38A-38H show line graphs representing bioluminescence intensity over time. These data are summarized in Figure 38I, which shows data for the first 30 days, and Figure 38J, which shows data for 56 days. Although Figure 38I shows a clear distinction between NK cells expressing the anti-CD19 CAR and controls, there is a nominal separation between the experimental groups. However, Figure 38J shows the 56-day data and there is a greater distinction in the ability of NK cells expressing the different CAR constructs and treated under the indicated conditions to inhibit tumor cell growth. It should be noted that “pairs” of groups (the same CAR construct, fresh and frozen) exhibit fairly similar anti-tumor activity. This indicates that, in several embodiments, engineered NK cells expressing the anti-CD19 CAR are not only effective when they are prepared and administered fresh. Additionally, in some embodiments, engineered NK cells expressing an anti-CD19 CAR are effective not only if they are obtained, frozen, then thawed and administered (e.g., as is the case with allogeneic cells).

[00313] Figure 39 shows a line graph of the body weight of mice treated with the constructs over a 56-day period of the experiment. The decrease in body weight correlates with increased tumor growth, such that tumor progression results in poor health of the mice and corresponding weight loss (e.g., wasting). As shown, the control groups exhibit significant weight loss over 30 days, while the experimental groups exhibited weight gain throughout most of the experiment. As with the bioluminescence data discussed above, there is a notable trend that many of the fresh and frozen preparations exhibited substantially similar effects on body weight when compared. In several embodiments, the engineered NK cells expressing an anti-CD19 CAR are effective not only when they are prepared and administered fresh. Additionally, in some embodiments, engineered NK cells expressing an anti-CD19 CAR are effective not only if they are obtained, frozen, then thawed and administered (e.g., as is the case with allogeneic cells).

[00314] It is contemplated that various combinations or subcombinations of the specific features and aspects of the embodiments disclosed above may be made and still fall within the scope of one or more of the disclosures of the present invention. Additionally, any particular feature, aspect, method, property, characteristic, quality, attribute, element or the like disclosed herein in connection with an embodiment may be applied to all other embodiments set forth herein. Accordingly, it should be understood that the various features and aspects of the disclosed embodiments may be combined or substituted for one another to produce various embodiments of the disclosures of the disclosed invention. Thus, it is not intended that the scope of the disclosures of the present invention disclosed herein should be limited to the specific disclosed embodiments described above. Moreover, although the present invention is susceptible to various modifications and alternative forms, specific examples thereof are shown in the drawings and described in detail herein. However, it should be understood that the present invention is not to be limited to the specific forms or methods disclosed, but on the contrary, the present invention is intended to cover all modifications, equivalents and alternatives falling within the spirit and scope of the various embodiments described and the appended claims. Any methods disclosed herein are not required to be performed in the order shown. The methods disclosed herein include specific actions taken by a practitioner; however, they may also include any third-party instructions for these actions, directly or indirectly. In addition, when features or aspects of the present invention are described in terms of Markush groups, those skilled in the art will understand that the invention so describes any individual member or subgroup of members of the Markush group.

[00315] The ranges disclosed herein also encompass any possible overlaps, subranges, and combinations thereof. Expressions such as "up to," "at least," "more than," "less than," "between," and the like include the stated number. Numbers preceded by a term such as "about" or "approximately" include the stated number. For example, "about 90%" includes "90%." In some embodiments, at least 95% sequence identity or homology includes 96%, 97%, 98%, 99%, and 100% sequence identity or homology to a reference sequence. Additionally, when a sequence is described as "comprising" a nucleotide or amino acid sequence, such reference shall also include, unless otherwise specified, that the sequence "comprises," "consists of," or "consists essentially of" the stated sequence.

[00316] Several embodiments provide amino acid sequences that correspond to any of the nucleic acids disclosed herein, taking into account the degeneracy of the nucleic acid code. Moreover, those sequences (whether a nucleic acid sequence or an amino acid sequence) that differ from those explicitly disclosed herein, but have functional similarity or equivalence, are also intended to be within the scope of the present invention. The foregoing includes mutants, truncations, substitutions, or other types of modifications.

[00317] Any headings or subheadings used herein are for organizational purposes and are not intended to limit the scope of the embodiments disclosed herein.

--->

SEQUENCE LIST

<110> Nkarta, Inc.

<120> CD19-DIRECTED CHIMERIC ANTIGEN RECEPTORS AND THEIR PATHWAYS

APPLICATIONS IN IMMUNOTHERAPY

<130>NKT.0033WO

<150> 62/814180

<151> 2019-03-05

<150> 62/895910

<151> 2019-09-04

<150> 62/932165

<151> 2019-11-07

<160> 207

<170> PatentIn version 3.5

<210> 1

<211> 135

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> CD8-alpha - DNA hinge

<400> 1

accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg 60

tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg 120

gacttcgcct gtgat 135

<210> 2

<211> 45

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CD8a hinge protein

<400> 2

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

1 5 10 15

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

20 25 30

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

35 40 45

<210> 3

<211> 63

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> TM CD8 - DNA

<400> 3

atctacatct gggcgccctt ggccgggact tgtggggtcc ttctcctgtc actggttatc 60

acc 63

<210> 4

<211> 21

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> TM CD8 - protein

<400> 4

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

1 5 10 15

Ser Leu Val Ile Thr

20

<210> 5

<211> 111

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> OX40 - DNA

<400> 5

cggagggacc agaggctgcc ccccgatgcc cacaagcccc ctgggggagg cagtttccgg 60

acccccatcc aagaggagca ggccgacgcc cactccaccc tggccaagat c 111

<210> 6

<211> 37

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> OX40 - protein

<400> 6

Arg Arg Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly

1 5 10 15

Gly Ser Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser

20 25 30

Th Leu Ala Lys Ile

35

<210> 7

<211> 336

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> CD3-zeta - DNA

<400> 7

agagtgaagt tcagcaggag cgcagacgcc cccgcgtacc agcagggcca gaaccagctc 60

tataacgagc tcaatctagg acgaagagag gagtacgatg ttttggacaa gagacgtggc 120

cgggaccctg agatgggggg aaagccgaga aggaagaacc ctcaggaagg cctgtacaat 180

gaactgcaga aagataagat ggcggaggcc tacagtgaga ttgggatgaa aggcgagcgc 240

cggaggggca aggggcacga tggcctttac cagggtctca gtacagccac caaggacacc 300

tacgacgccc ttcacatgca ggccctgccc cctcgc 336

<210> 8

<211> 112

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CD3-zeta protein

<400> 8

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly

1 5 10 15

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

20 25 30

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

35 40 45

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

50 55 60

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

65 70 75 80

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

85 90 95

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

100 105 110

<210> 9

<211> 63

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> T2A - DNA

<400> 9

ggctctggcg agggaagggg ttccctgctt acttgcggcg acgtcgaaga gaatcccggt 60

ccg 63

<210> 10

<211> 21

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> T2A - protein

<400> 10

Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu

1 5 10 15

Glu Asn Pro Gly Pro

20

<210> 11

<211> 342

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> IL15 - DNA

<400> 11

aactgggtca acgtgattag cgatttgaag aaaatcgagg accttataca gtctatgcat 60

attgacgcta cactgtatac tgagagtgat gtacacccgt cctgtaaggt aacggccatg 120

aaatgctttc ttctggagct ccaggtcatc agcttggagt ctggggacgc aagcatccac 180

gatacggttg aaaacctcat catccttgcg aacaactctc tctcatctaa tggaaacgtt 240

acagagagtg ggtgtaagga gtgcgaagag ttggaagaaa aaaacatcaa agaatttctt 300

caatccttcg ttcacatagt gcaaatgttc attaacacgt cc 342

<210> 12

<211> 114

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> IL15 - protein

<400> 12

Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile

1 5 10 15

Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His

20 25 30

Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln

35 40 45

Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu

50 55 60

Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val

65 70 75 80

Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile

85 90 95

Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn

100 105 110

Thr Ser

<210> 13

<211> 207

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Hinge/TM CD8 - DNA

<400> 13

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 60

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 120

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 180

ctctcactcg ttattacgct gtactgc 207

<210> 14

<211> 66

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Hinge/TM CD8 - protein

<400> 14

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

1 5 10 15

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

20 25 30

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile

35 40 45

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

50 55 60

Ile Th

65

<210> 15

<211> 30

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Linker 1 (used after FLAG tag) - DNA

<400> 15

ggcggtggtg gctctggtgg tggcggcagc 30

<210> 16

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Linker 1 (used after the FLAG tag) - protein

<400> 16

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10

<210> 17

<211> 30

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Linker 2 - after the negative control binding domain - DNA

<400> 17

ggccaggccg gctccggagg aggaggatcc 30

<210> 18

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Linker 2 - after the negative control binding domain - protein

<400> 18

Gly Gln Ala Gly Ser Gly Gly Gly Gly Ser

1 5 10

<210> 19

<211> 12

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Linker after scFv - DNA

<400> 19

ggccaggccg gc 12

<210> 20

<211> 4

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Linker after scFv - protein

<400> 20

Gly Gln Ala Gly

1

<210> 21

<211> 33

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Linker - DNA

<220>

<221> another sign

<223> Another linker is DNA

<400> 21

ggcggcggcg gtagcggtgg tggcggctcc gga 33

<210> 22

<211> 11

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Another linker is a protein

<400> 22

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

1 5 10

<210> 23

<211> 18

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Linker - DNA

<400> 23

ggccaggccg gctccgga 18

<210> 24

<211> 6

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Linker - protein

<400> 24

Gly Gln Ala Gly Ser Gly

1 5

<210> 25

<211> 405

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Extracellular fragment of NKG2D - DNA

<400> 25

ttattcaacc aagaagttca aattcccttg accgaaagtt actgtggccc atgtcctaaa 60

aactggatat gttacaaaaa taactgctac caattttttg atgagagtaa aaactggtat 120

gagagccagg cttcttgtat gtctcaaaat gccagccttc tgaaagtata cagcaaagag 180

gaccaggatt tacttaaact ggtgaagtca tatcattgga tgggactagt acacattcca 240

acaaatggat cttggcagtg ggaagatggc tccattctct cacccaacct actaacaata 300

attgaaatgc agaagggaga ctgtgcactc tatgcctcga gctttaaagg ctatatagaa 360

aactgttcaa ctccaaatac gtacatctgc atgcaaagga ctgtg 405

<210> 26

<211> 135

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Extracellular fragment of NKG2D - protein

<400> 26

Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys Gly

1 5 10 15

Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln Phe

20 25 30

Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met Ser

35 40 45

Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp Leu

50 55 60

Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile Pro

65 70 75 80

Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro Asn

85 90 95

Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr Ala

100 105 110

Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro Asn Thr Tyr

115 120 125

Ile Cys Met Gln Arg Thr Val

130 135

<210> 27

<211> 645

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Whole human NKG2D - DNA

<400> 27

gggtggattc gtggtcggag gtctcgacac agctgggaga tgagtgaatt tcataattat 60

aacttggatc tgaagaagag tgatttttca acacgatggc aaaagcaaag atgtccagta 120

gtcaaaagca aatgtagaga aaatgcatct ccattttttt tctgctgctt catcgctgta 180

gccatgggaa tccgtttcat tattatggta acaatatgga gtgctgtatt cctaaactca 240

ttattcaacc aagaagttca aattcccttg accgaaagtt actgtggccc atgtcctaaa 300

aactggatat gttacaaaaa taactgctac caattttttg atgagagtaa aaactggtat 360

gagagccagg cttcttgtat gtctcaaaat gccagccttc tgaaagtata cagcaaagag 420

gaccaggatt tacttaaact ggtgaagtca tatcattgga tgggactagt acacattcca 480

acaaatggat cttggcagtg ggaagatggc tccattctct cacccaacct actaacaata 540

attgaaatgc agaagggaga ctgtgcactc tatgcctcga gctttaaagg ctatatagaa 600

aactgttcaa ctccaaatac gtacatctgc atgcaaagga ctgtg 645

<210> 28

<211> 405

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Codon-optimized fragment of human NKG2D - DNA

<400> 28

ctgttcaatc aggaagtcca gatccccctg acagagtctt actgcggccc atgtcccaag 60

aactggatct gctacaagaa caattgttat cagttctttg acgagagcaa gaactggtat 120

gagtcccagg cctcttgcat gagccagaat gcctctctgc tgaaggtgta cagcaaggag 180

gaccaggatc tgctgaagct ggtgaagtcc tatcactgga tgggcctggt gcacatccct 240

acaaacggct cttggcagtg ggaggacggc tccatcctgt ctccaaatct gctgaccatc 300

atcgagatgc agaagggcga ttgcgccctg tacgccagct ccttcaaggg ctatatcgag 360

aactgctcca cacccaatac ctacatctgt atgcagagga ccgtg 405

<210> 29

<211> 126

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> 4-1BB

<400> 29

aaacggggca gaaagaaact cctgtatata ttcaaacaac catttatgag accagtacaa 60

actactcaag aggaagatgg ctgtagctgc cgatttccag aagaagaaga aggaggatgt 120

gaactg 126

<210> 30

<211> 29

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Amino acid sequence of the transmembrane domain of CD28

<400> 30

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu

1 5 10 15

Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser

20 25

<210> 31

<211> 39

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Amino acid sequence of the IC domain of CD28

<400> 31

Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg

1 5 10 15

Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg

20 25 30

Asp Phe Ala Ala Tyr Arg Ser

35

<210> 32

<211> 123

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> V. L. Nicholson et al.

<400> 32

Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr Arg Ala Asp Ala Ala

100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Asn

115 120

<210> 33

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> V. H. Nicholson et al.

<400> 33

Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln

1 5 10 15

Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu

65 70 75 80

Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 34

<211> 634

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Protein artificial sequence of chimeric receptor CD19R-zeta

<400> 34

Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro

1 5 10 15

Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser

20 25 30

Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser

35 40 45

Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly

50 55 60

Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val

65 70 75 80

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr

85 90 95

Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln

100 105 110

Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile

115 120 125

Thr Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser

130 135 140

Thr Lys Gly Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala

145 150 155 160

Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu

165 170 175

Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu

180 185 190

Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser

195 200 205

Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln

210 215 220

Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr

225 230 235 240

Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

245 250 255

Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Val Glu Pro Lys Ser

260 265 270

Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

275 280 285

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

290 295 300

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

305 310 315 320

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

325 330 335

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

340 345 350

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

355 360 365

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

370 375 380

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

385 390 395 400

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

405 410 415

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

420 425 430

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

435 440 445

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

450 455 460

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

465 470 475 480

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

485 490 495

Ser Pro Gly Lys Met Ala Leu Ile Val Leu Gly Gly Val Ala Gly Leu

500 505 510

Leu Leu Phe Ile Gly Leu Gly Ile Phe Phe Arg Val Lys Phe Ser Arg

515 520 525

Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn

530 535 540

Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg

545 550 555 560

Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro

565 570 575

Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala

580 585 590

Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His

595 600 605

Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp

610 615 620

Ala Leu His Met Gln Ala Leu Pro Pro Arg

625 630

<210> 35

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> HCV scFv to CD19

<400> 35

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 36

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> HCV scFv to CD19

<400> 36

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 37

<211> 11

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 LC

<400> 37

Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn

1 5 10

<210> 38

<211> 7

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 LC

<400> 38

His Thr Ser Arg Leu His Ser

1 5

<210> 39

<211> 9

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 LC

<400> 39

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

1 5

<210> 40

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1HC

<400> 40

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 41

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2HC

<400> 41

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Ser Ser Leu Lys Ser

1 5 10 15

<210> 42

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HC - alternative variant

<400> 42

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Gln Ser Ser Leu Lys Ser

1 5 10 15

<210> 43

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HC - alternative variant 2

<400> 43

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ser Leu Lys Ser

1 5 10 15

<210> 44

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3HC

<400> 44

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 45

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Light chain variable region 1

<400> 45

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gln Ala Pro Lys Leu Leu Ile

35 40 45

Lys His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 46

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Light chain variable region 2

<400> 46

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 47

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Light chain variable region 3

<400> 47

Asp Ile Gln Met Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gln Ala Pro Lys Leu Leu Ile

35 40 45

Lys His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 48

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Light chain variable region 4

<400> 48

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gln Ala Pro Lys Leu Leu Ile

35 40 45

Lys His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 49

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain variable region 1

<400> 49

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 50

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain variable region 2

<400> 50

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Phe Leu

65 70 75 80

Lys Met Ser Ser Leu Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 51

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain variable region 3

<400> 51

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 52

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain variable region 4

<400> 52

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 53

<211> 8

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 LC

<400> 53

Arg Ala Ser Gln Asp Ile Ser Lys

1 5

<210> 54

<211> 7

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 LC

<400> 54

Ile Tyr His Thr Ser Arg Leu

1 5

<210> 55

<211> 9

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 LC

<400> 55

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

1 5

<210> 56

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1HC

<400> 56

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 57

<211> 14

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2HC

<400> 57

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

1 5 10

<210> 58

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3HC

<400> 58

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 59

<211> 2210

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-1b DNA

<400> 59

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1080

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1140

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1200

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1260

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1320

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1380

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1440

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1500

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1560

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1620

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1680

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1740

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1800

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1860

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1920

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1980

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 2040

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2100

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2160

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2210

<210> 60

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-1b protein

<400> 60

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 61

<211> 2231

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-2b DNA

<400> 61

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgatt tttgggtgct ggtggtggtt 1020

ggtggagtcc tggcttgcta tagcttgcta gtaacagtgg cctttattat tttctgggtg 1080

aggagtaaga ggagcaggct cctgcacagt gactacatga acatgactcc ccgccgcccc 1140

gggcccaccc gcaagcatta ccagccctat gccccaccac gcgacttcgc agcctatcgc 1200

tccagagtga agttcagcag gagcgcagac gcccccgcgt accagcaggg ccagaaccag 1260

ctctataacg agctcaatct aggacgaaga gaggagtacg atgttttgga caagagacgt 1320

ggccgggacc ctgagatggg gggaaagccg agaaggaaga accctcagga aggcctgtac 1380

aatgaactgc agaaagataa gatggcggag gcctacagtg agattgggat gaaaggcgag 1440

cgccggaggg gcaaggggca cgatggcctt taccagggtc tcagtacagc caccaaggac 1500

acctacgacg cccttcacat gcaggccctg ccccctcgcg gctctggcga gggaaggggt 1560

tccctgctta cttgcggcga cgtcgaagag aatcccggtc cgatggccct cccagtaact 1620

gccctccttt tgcccctcgc actccttctt catgccgctc gccccaactg ggtcaacgtg 1680

attagcgatt tgaagaaaat cgaggacctt atacagtcta tgcatattga cgctacactg 1740

tatactgaga gtgatgtaca cccgtcctgt aaggtaacgg ccatgaaatg ctttcttctg 1800

gagctccagg tcatcagctt ggagtctggg gacgcaagca tccacgatac ggttgaaaac 1860

ctcatcatcc ttgcgaacaa ctctctctca tctaatggaa acgttacaga gagtgggtgt 1920

aaggagtgcg aagagttgga agaaaaaaac atcaaagaat ttcttcaatc cttcgttcac 1980

atagtgcaaa tgttcattaa cacgtccact accacacccg ccccgaggcc acctacgccg 2040

gcaccgacta tcgccagtca acccctctct ctgcgccccg aggcttgccg gcctgcggct 2100

ggtggggcgg tccacacccg gggcctggat tttgcgtgcg atatatacat ctgggcacct 2160

cttgccggca cctgcggagt gctgcttctc tcactcgtta ttacgctgta ctgctaagcg 2220

gccgcgtcga c 2231

<210> 62

<211> 731

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-2b protein

<400> 62

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Phe Trp Val Leu Val Val Val Gly Gly Val

325 330 335

Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp

340 345 350

Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met

355 360 365

Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala

370 375 380

Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg

385 390 395 400

Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn

405 410 415

Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg

420 425 430

Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro

435 440 445

Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala

450 455 460

Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His

465 470 475 480

Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp

485 490 495

Ala Leu His Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg

500 505 510

Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met

515 520 525

Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu His

530 535 540

Ala Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile

545 550 555 560

Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu

565 570 575

Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu

580 585 590

Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His

595 600 605

Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser

610 615 620

Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu

625 630 635 640

Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln

645 650 655

Met Phe Ile Asn Thr Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr

660 665 670

Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala

675 680 685

Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe

690 695 700

Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val

705 710 715 720

Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

725 730

<210> 63

<211> 2213

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-3b DNA

<400> 63

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ctgttggctt 1080

acaaaaaaga agtattcatc cagtgtgcac gaccctaacg gtgaatacat gttcatgaga 1140

gcagtgaaca cagccaaaaa atctagactc acagatgtga ccctaagagt gaagttcagc 1200

aggagcgcag acgcccccgc gtaccagcag ggccagaacc agctctataa cgagctcaat 1260

ctaggacgaa gagaggagta cgatgttttg gacaagagac gtggccggga ccctgagatg 1320

gggggaaagc cgagaaggaa gaaccctcag gaaggcctgt acaatgaact gcagaaagat 1380

aagatggcgg aggcctacag tgagattggg atgaaaggcg agcgccggag gggcaagggg 1440

cacgatggcc tttaccaggg tctcagtaca gccaccaagg acacctacga cgcccttcac 1500

atgcaggccc tgccccctcg cggctctggc gagggaaggg gttccctgct tacttgcggc 1560

gacgtcgaag agaatcccgg tccgatggcc ctcccagtaa ctgccctcct tttgcccctc 1620

gcactccttc ttcatgccgc tcgccccaac tgggtcaacg tgattagcga tttgaagaaa 1680

atcgaggacc ttatacagtc tatgcatatt gacgctacac tgtatactga gagtgatgta 1740

cacccgtcct gtaaggtaac ggccatgaaa tgctttcttc tggagctcca ggtcatcagc 1800

ttggagtctg gggacgcaag catccacgat acggttgaaa acctcatcat ccttgcgaac 1860

aactctctct catctaatgg aaacgttaca gagagtgggt gtaaggagtg cgaagagttg 1920

gaagaaaaaa acatcaaaga atttcttcaa tccttcgttc acatagtgca aatgttcatt 1980

aacacgtcca ctaccacacc cgccccgagg ccacctacgc cggcaccgac tatcgccagt 2040

caacccctct ctctgcgccc cgaggcttgc cggcctgcgg ctggtggggc ggtccacacc 2100

cggggcctgg attttgcgtg cgatatatac atctgggcac ctcttgccgg cacctgcgga 2160

gtgctgcttc tctcactcgt tattacgctg tactgctaag cggccgcgtc gac 2213

<210> 64

<211> 725

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-3b protein

<400> 64

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Cys Trp

340 345 350

Leu Thr Lys Lys Lys Tyr Ser Ser Ser Val His Asp Pro Asn Gly Glu

355 360 365

Tyr Met Phe Met Arg Ala Val Asn Thr Ala Lys Lys Ser Arg Leu Thr

370 375 380

Asp Val Thr Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala

385 390 395 400

Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg

405 410 415

Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu

420 425 430

Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn

435 440 445

Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met

450 455 460

Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly

465 470 475 480

Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala

485 490 495

Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys

500 505 510

Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala

515 520 525

Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp

530 535 540

Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser

545 550 555 560

Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser

565 570 575

Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile

580 585 590

Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu

595 600 605

Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu

610 615 620

Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu

625 630 635 640

Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser

645 650 655

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

660 665 670

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

675 680 685

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile

690 695 700

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

705 710 715 720

Ile Thr Leu Tyr Cys

725

<210> 65

<211> 2357

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-4b DNA

<400> 65

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actgtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgatt tttgggtgct ggtggtggtt 1020

ggtggagtcc tggcttgcta tagcttgcta gtaacagtgg cctttattat tttctgggtg 1080

aggagtaaga ggagcaggct cctgcacagt gactacatga acatgactcc ccgccgcccc 1140

gggcccaccc gcaagcatta ccagccctat gccccaccac gcgacttcgc agcctatcgc 1200

tccaaacggg gcagaaagaa actcctgtat atattcaaac aaccatttat gagaccagta 1260

caaactactc aagaggaaga tggctgtagc tgccgatttc cagaagaaga agaagggagga 1320

tgtgaactga gagtgaagtt cagcaggagc gcagacgccc ccgcgtacca gcagggccag 1380

aaccagctct ataacgagct caatctagga cgaagagagg agtacgatgt tttggacaag 1440

agacgtggcc gggaccctga gatgggggga aagccgagaa ggaagaaccc tcaggaaggc 1500

ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa 1560

ggcgagcgcc ggaggggcaa ggggcacgat ggcctttacc agggtctcag tacagccacc 1620

aaggacacct acgacgccct tcacatgcag gccctgcccc ctcgcggctc tggcgaggga 1680

aggggttccc tgcttacttg cggcgacgtc gaagagaatc ccggtccgat ggccctccca 1740

gtaactgccc tccttttgcc cctcgcactc cttcttcatg ccgctcgccc caactgggtc 1800

aacgtgatta gcgatttgaa gaaaatcgag gaccttatac agtctatgca tattgacgct 1860

acactgtata ctgagagtga tgtacacccg tcctgtaagg taacggccat gaaatgcttt 1920

cttctggagc tccaggtcat cagcttggag tctggggacg caagcatcca cgatacggtt 1980

gaaaacctca tcatccttgc gaacaactct ctctcatcta atggaaacgt tacagagagt 2040

gggtgtaagg agtgcgaaga gttggaagaa aaaaacatca aagaatttct tcaatccttc 2100

gttcacatag tgcaaatgtt cattaacacg tccactacca cacccgcccc gaggccacct 2160

acgccggcac cgactatcgc cagtcaaccc ctctctctgc gccccgaggc ttgccggcct 2220

gcggctggtg gggcggtcca cacccggggc ctggattttg cgtgcgatat atacatctgg 2280

gcacctcttg ccggcacctg cggagtgctg cttctctcac tcgttattac gctgtactgc 2340

taagcggccg cgtcgac 2357

<210> 66

<211> 773

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-4b protein

<400> 66

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Phe Trp Val Leu Val Val Val Gly Gly Val

325 330 335

Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp

340 345 350

Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met

355 360 365

Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala

370 375 380

Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Lys Arg Gly Arg Lys Lys

385 390 395 400

Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr

405 410 415

Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly

420 425 430

Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala

435 440 445

Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg

450 455 460

Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu

465 470 475 480

Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn

485 490 495

Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met

500 505 510

Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly

515 520 525

Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala

530 535 540

Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys

545 550 555 560

Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala

565 570 575

Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp

580 585 590

Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser

595 600 605

Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser

610 615 620

Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile

625 630 635 640

Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu

645 650 655

Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu

660 665 670

Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu

675 680 685

Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser

690 695 700

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

705 710 715 720

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

725 730 735

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile

740 745 750

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

755 760 765

Ile Thr Leu Tyr Cys

770

<210> 67

<211> 2201

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-5b DNA

<400> 67

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgatc catttttttt ctgctgcttc 1020

atcgctgtag ccatgggaat ccgtttcatt attatggtaa cacggaggga ccagaggctg 1080

ccccccgatg cccacaagcc ccctggggga ggcagtttcc ggacccccat ccaagaggag 1140

caggccgacg cccactccac cctggccaag atcagagtga agttcagcag gagcgcagac 1200

gcccccgcgt accagcaggg ccagaaccag ctctataacg agctcaatct aggacgaaga 1260

gaggagtacg atgttttgga caagagacgt ggccgggacc ctgagatggg gggaaagccg 1320

agaaggaaga accctcagga aggcctgtac aatgaactgc agaaagataa gatggcggag 1380

gcctacagtg agattgggat gaaaggcgag cgccggaggg gcaaggggca cgatggcctt 1440

taccagggtc tcagtacagc caccaaggac acctacgacg cccttcacat gcaggccctg 1500

ccccctcgcg gctctggcga gggaaggggt tccctgctta cttgcggcga cgtcgaagag 1560

aatcccggtc cgatggccct cccagtaact gccctccttt tgcccctcgc actccttctt 1620

catgccgctc gccccaactg ggtcaacgtg attagcgatt tgaagaaaat cgaggacctt 1680

atacagtcta tgcatattga cgctacactg tatactgaga gtgatgtaca cccgtcctgt 1740

aaggtaacgg ccatgaaatg ctttcttctg gagctccagg tcatcagctt ggagtctggg 1800

gacgcaagca tccacgatac ggttgaaaac ctcatcatcc ttgcgaacaa ctctctctca 1860

tctaatggaa acgttacaga gagtgggtgt aaggagtgcg aagagttgga agaaaaaaac 1920

atcaaagaat ttcttcaatc cttcgttcac atagtgcaaa tgttcattaa cacgtccact 1980

accacacccg ccccgaggcc acctacgccg gcaccgacta tcgccagtca acccctctct 2040

ctgcgccccg aggcttgccg gcctgcggct ggtggggcgg tccacacccg gggcctggat 2100

tttgcgtgcg atatatacat ctgggcacct cttgccggca cctgcggagt gctgcttctc 2160

tcactcgtta ttacgctgta ctgctaagcg gccgcgtcga c 2201

<210> 68

<211> 721

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-5b protein

<400> 68

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Pro Phe Phe Phe Cys Cys Phe Ile Ala Val

325 330 335

Ala Met Gly Ile Arg Phe Ile Ile Met Val Thr Arg Arg Asp Gln Arg

340 345 350

Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg Thr

355 360 365

Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys Ile

370 375 380

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly

385 390 395 400

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

405 410 415

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

420 425 430

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

435 440 445

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

450 455 460

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

465 470 475 480

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

485 490 495

Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu

500 505 510

Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro

515 520 525

Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val Ile

530 535 540

Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp

545 550 555 560

Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr

565 570 575

Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser

580 585 590

Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala

595 600 605

Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys

610 615 620

Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser

625 630 635 640

Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr Pro

645 650 655

Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu

660 665 670

Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His

675 680 685

Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu

690 695 700

Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr

705 710 715 720

Cys

<210> 69

<211> 1805

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-6b DNA

<400> 69

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacaa ccacgacgcc agcgccgcga 480

ccaccaacac cggcgcccac catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc 540

cggccagcgg cggggggcgc agtgcacacg agggggctgg acttcgcctg tgatatctac 600

atctgggcgc ccttggccgg gacttgtggg gtccttctcc tgtcactggt tatcaccctt 660

tactgccgga gggaccagag gctgcccccc gatgcccaca agccccctgg gggaggcagt 720

ttccggaccc ccatccaaga ggagcaggcc gacgcccact ccaccctggc caagatcaga 780

gtgaagttca gcaggagcgc agacgccccc gcgtaccagc agggccagaa ccagctctat 840

aacgagctca atctaggacg aagagaggag tacgatgttt tggacaagag acgtggccgg 900

gaccctgaga tggggggaaa gccgagaagg aagaaccctc aggaaggcct gtacaatgaa 960

ctgcagaaag ataagatggc ggaggcctac agtgagattg ggatgaaagg cgagcgccgg 1020

aggggcaagg ggcacgatgg cctttaccag ggtctcagta cagccaccaa ggacacctac 1080

gacgcccttc acatgcaggc cctgccccct cgcggctctg gcgagggaag gggttccctg 1140

cttacttgcg gcgacgtcga agagaatccc ggtccgatgg ccctcccagt aactgccctc 1200

cttttgcccc tcgcactcct tcttcatgcc gctcgcccca actgggtcaa cgtgattagc 1260

gatttgaaga aaatcgagga ccttatacag tctatgcata ttgacgctac actgtatact 1320

gagagtgatg tacacccgtc ctgtaaggta acggccatga aatgctttct tctggagctc 1380

caggtcatca gcttggagtc tggggacgca agcatccacg atacggttga aaacctcatc 1440

atccttgcga acaactctct ctcatctaat ggaaacgtta cagagagtgg gtgtaaggag 1500

tgcgaagagt tggaagaaaa aaacatcaaa gaatttcttc aatccttcgt tcacatagtg 1560

caaatgttca ttaacacgtc cactaccaca cccgccccga ggccacctac gccggcaccg 1620

actatcgcca gtcaacccct ctctctgcgc cccgaggctt gccggcctgc ggctggtggg 1680

gcggtccaca cccggggcct ggattttgcg tgcgatatat acatctgggc acctcttgcc 1740

ggcacctgcg gagtgctgct tctctcactc gttattacgc tgtactgcta agcggccgcg 1800

tcgac 1805

<210> 70

<211> 589

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-6b protein

<400> 70

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr

145 150 155 160

Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala

165 170 175

Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile

180 185 190

Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser

195 200 205

Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln Arg Leu Pro Pro Asp

210 215 220

Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg Thr Pro Ile Gln Glu

225 230 235 240

Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys Ile Arg Val Lys Phe

245 250 255

Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu

260 265 270

Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp

275 280 285

Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys

290 295 300

Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala

305 310 315 320

Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys

325 330 335

Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr

340 345 350

Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Glu

355 360 365

Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly

370 375 380

Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu

385 390 395 400

Leu His Ala Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys

405 410 415

Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr

420 425 430

Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys

435 440 445

Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser

450 455 460

Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu

465 470 475 480

Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu

485 490 495

Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile

500 505 510

Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr Pro Ala Pro Arg Pro

515 520 525

Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro

530 535 540

Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu

545 550 555 560

Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys

565 570 575

Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

580 585

<210> 71

<211> 3347

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-7b DNA

<400> 71

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1080

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1140

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1200

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1260

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1320

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1380

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1440

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1500

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1560

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1620

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1680

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1740

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1800

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1860

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1920

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1980

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 2040

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2100

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2160

ctgcttctct cactcgttat tacgctgtac tgcggcagcg gcgccacaaa cttctctctg 2220

ctaaagcaag caggtgatgt tgaagaaaac cccggggccta tgcttctcct ggtgacaagc 2280

cttctgctct gtgagttacc acacccagca ttcctcctga tcccacgcaa agtgtgtaac 2340

ggaataggta ttggtgaatt taaagactca ctctccataa atgctacgaa tattaaacac 2400

ttcaaaaact gcacctccat cagtggcgat ctccacatcc tgccggtggc atttaggggt 2460

gactccttca cacatactcc tcctctggat ccacaggaac tggatattct gaaaaccgta 2520

aaggaaatca cagggttttt gctgattcag gcttggcctg aaaacaggac ggacctccat 2580

gcctttgaga acctagaaat catacgcggc aggaccaagc aacatggtca gttttctctt 2640

gcagtcgtca gcctgaacat aacatccttg ggattacgct ccctcaagga gataagtgat 2700

ggagatgtga taatttcagg aaacaaaaat ttgtgctatg caaatacaat aaactggaaa 2760

aaactgtttg ggacctccgg tcagaaaacc aaaattataa gcaacagagg tgaaaacagc 2820

tgcaaggcca caggccaggt ctgccatgcc ttgtgctccc ccgagggctg ctggggcccg 2880

gagcccaggg actgcgtctc ttgccggaat gtcagccgag gcagggaatg cgtggacaag 2940

tgcaaccttc tggagggtga gccaagggag tttgtggaga actctgagtg catacagtgc 3000

cacccagagt gcctgcctca ggccatgaac atcacctgca caggacgggg accagacaac 3060

tgtatccagt gtgcccacta cattgacggc ccccactgcg tcaagacctg cccggcagga 3120

gtcatgggag aaaacaacac cctggtctgg aagtacgcag acgccggcca tgtgtgccac 3180

ctgtgccatc caaactgcac ctacggatgc actgggccag gtcttgaagg ctgtccaacg 3240

aatgggccta agatcccgtc catcgccact gggatggtgg gggccctcct cttgctgctg 3300

gtggtggccc tggggatcgg cctcttcatg tgagcggccg cgtcgac 3347

<210> 72

<211> 1103

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-7b protein

<400> 72

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln

725 730 735

Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Met Leu Leu Leu Val Thr

740 745 750

Ser Leu Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu Leu Ile Pro

755 760 765

Arg Lys Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu

770 775 780

Ser Ile Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile

785 790 795 800

Ser Gly Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe

805 810 815

Thr His Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr

820 825 830

Val Lys Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn

835 840 845

Arg Thr Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg

850 855 860

Thr Lys Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile

865 870 875 880

Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val

885 890 895

Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp

900 905 910

Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn

915 920 925

Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu

930 935 940

Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser

945 950 955 960

Cys Arg Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys Cys Asn Leu

965 970 975

Leu Glu Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu Cys Ile Gln

980 985 990

Cys His Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr Cys Thr Gly

995 1000 1005

Arg Gly Pro Asp Asn Cys Ile Gln Cys Ala His Tyr Ile Asp Gly

1010 1015 1020

Pro His Cys Val Lys Thr Cys Pro Ala Gly Val Met Gly Glu Asn

1025 1030 1035

Asn Thr Leu Val Trp Lys Tyr Ala Asp Ala Gly His Val Cys His

1040 1045 1050

Leu Cys His Pro Asn Cys Thr Tyr Gly Cys Thr Gly Pro Gly Leu

1055 1060 1065

Glu Gly Cys Pro Thr Asn Gly Pro Lys Ile Pro Ser Ile Ala Thr

1070 1075 1080

Gly Met Val Gly Ala Leu Leu Leu Leu Leu Val Val Ala Leu Gly

1085 1090 1095

Ile Gly Leu Phe Met

1100

<210> 73

<211> 1844

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-8b DNA

<400> 73

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga ggtgaaactg caggagtcag gacctggcct ggtggcgccc 180

tcacagagcc tgtccgtcac atgcactgtc tcaggggtct cattacccga ctatggtgta 240

agctggattc gccagcctcc acgaaagggt ctggagtggc tgggagtaat atggggtagt 300

gaaaccacat actataattc agctctcaaa tccagactga ccatcatcaa ggacaactcc 360

aagagccaag ttttcttaaa aatgaacagt ctgcaaactg atgacacagc catttactac 420

tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg ccaaggaacc 480

tcagtcaccg tctcctcaac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 540

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 600

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 660

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 720

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 780

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 840

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 900

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 960

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1020

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1080

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1140

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1200

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1260

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1320

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1380

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1440

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1500

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1560

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1620

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 1680

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 1740

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 1800

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 1844

<210> 74

<211> 602

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-8b protein

<400> 74

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Glu Val Lys Leu Gln Glu Ser Gly Pro

35 40 45

Gly Leu Val Ala Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser

50 55 60

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro

65 70 75 80

Arg Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr

85 90 95

Tyr Tyr Asn Ser Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn

100 105 110

Ser Lys Ser Gln Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp

115 120 125

Thr Ala Ile Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr

130 135 140

Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Thr

145 150 155 160

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

165 170 175

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

180 185 190

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

195 200 205

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

210 215 220

Thr Leu Tyr Cys Arg Arg Asp Gln Arg Leu Pro Pro Asp Ala His Lys

225 230 235 240

Pro Pro Gly Gly Gly Ser Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala

245 250 255

Asp Ala His Ser Thr Leu Ala Lys Ile Arg Val Lys Phe Ser Arg Ser

260 265 270

Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu

275 280 285

Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg

290 295 300

Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln

305 310 315 320

Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr

325 330 335

Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp

340 345 350

Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala

355 360 365

Leu His Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly

370 375 380

Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala

385 390 395 400

Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala

405 410 415

Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu

420 425 430

Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser

435 440 445

Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu

450 455 460

Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp

465 470 475 480

Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn

485 490 495

Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu

500 505 510

Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met

515 520 525

Phe Ile Asn Thr Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro

530 535 540

Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys

545 550 555 560

Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala

565 570 575

Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu

580 585 590

Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

595 600

<210> 75

<211> 2243

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-9b DNA

<400> 75

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccaacgaagg 1080

aaatatagat caaacaaagg agaaagtcct gtggagcctg cagagccttg tcgttacagc 1140

tgccccaggg aggagggag cagcaccatc cccatccagg aggattaccg aaaaccggag 1200

cctgcctgct cccccagagt gaagttcagc aggagcgcag acgcccccgc gtaccagcag 1260

ggccagaacc agctctataa cgagctcaat ctaggacgaa gagaggagta cgatgttttg 1320

gacaagagac gtggccggga ccctgagatg gggggaaagc cgagaaggaa gaaccctcag 1380

gaaggcctgt acaatgaact gcagaaagat aagatggcgg aggcctacag tgagattggg 1440

atgaaaggcg agcgccggag gggcaagggg cacgatggcc tttaccaggg tctcagtaca 1500

gccaccaagg acacctacga cgcccttcac atgcaggccc tgccccctcg cggctctggc 1560

gagggaaggg gttccctgct tacttgcggc gacgtcgaag agaatcccgg tccgatggcc 1620

ctcccagtaa ctgccctcct tttgcccctc gcactccttc ttcatgccgc tcgccccaac 1680

tgggtcaacg tgattagcga tttgaagaaa atcgaggacc ttatacagtc tatgcatatt 1740

gacgctacac tgtatactga gagtgatgta cacccgtcct gtaaggtaac ggccatgaaa 1800

tgctttcttc tggagctcca ggtcatcagc ttggagtctg gggacgcaag catccacgat 1860

acggttgaaa acctcatcat ccttgcgaac aactctctct catctaatgg aaacgttaca 1920

gagagtgggt gtaaggagtg cgaagagttg gaagaaaaaa acatcaaaga atttcttcaa 1980

tccttcgttc acatagtgca aatgttcatt aacacgtcca ctaccacacc cgccccgagg 2040

ccacctacgc cggcaccgac tatcgccagt caacccctct ctctgcgccc cgaggcttgc 2100

cggcctgcgg ctggtggggc ggtccacacc cggggcctgg attttgcgtg cgatatatac 2160

atctgggcac ctcttgccgg cacctgcgga gtgctgcttc tctcactcgt tattacgctg 2220

tactgctaag cggccgcgtc gac 2243

<210> 76

<211> 735

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-9b protein

<400> 76

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Gln Arg

340 345 350

Arg Lys Tyr Arg Ser Asn Lys Gly Glu Ser Pro Val Glu Pro Ala Glu

355 360 365

Pro Cys Arg Tyr Ser Cys Pro Arg Glu Glu Glu Gly Ser Thr Ile Pro

370 375 380

Ile Gln Glu Asp Tyr Arg Lys Pro Glu Pro Ala Cys Ser Pro Arg Val

385 390 395 400

Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn

405 410 415

Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val

420 425 430

Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg

435 440 445

Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys

450 455 460

Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg

465 470 475 480

Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys

485 490 495

Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Gly Ser

500 505 510

Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn

515 520 525

Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala

530 535 540

Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp

545 550 555 560

Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr

565 570 575

Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met

580 585 590

Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp

595 600 605

Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn

610 615 620

Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys

625 630 635 640

Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val

645 650 655

His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr Pro Ala Pro

660 665 670

Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu

675 680 685

Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg

690 695 700

Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly

705 710 715 720

Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

725 730 735

<210> 77

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-10b DNA

<400> 77

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actgtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg catgccggag 1080

gagggttcgg gctgctcggt gcggcgcagg ccctatgggt gcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 78

<211> 704

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-10b protein

<400> 78

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Met Pro

340 345 350

Glu Glu Gly Ser Gly Cys Ser Val Arg Arg Arg Pro Tyr Gly Cys Arg

355 360 365

Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln

370 375 380

Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp

385 390 395 400

Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro

405 410 415

Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp

420 425 430

Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg

435 440 445

Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr

450 455 460

Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Gly

465 470 475 480

Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu

485 490 495

Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu

500 505 510

Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val Ile Ser

515 520 525

Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala

530 535 540

Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala

545 550 555 560

Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly

565 570 575

Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn

580 585 590

Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu

595 600 605

Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe

610 615 620

Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr Pro Ala

625 630 635 640

Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser

645 650 655

Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr

660 665 670

Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala

675 680 685

Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

690 695 700

<210> 79

<211> 2234

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-11b DNA

<400> 79

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg catggaccaa 1080

caagcaatat atgctgagtt aaacttaccc acagactcag gcccagaaag ttcttcacct 1140

tcatctcttc ctcgggatgt ctgtcagggt tcaccttggc atcaatttgc cctgaaactt 1200

agctgtagag tgaagttcag caggagcgca gacgcccccg cgtaccagca gggccagaac 1260

cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga 1320

cgtggccggg accctgagat ggggggaaag ccgagaagga agaaccctca ggaaggcctg 1380

tacaatgaac tgcagaaaga taagatggcg gaggcctaca gtgagattgg gatgaaaggc 1440

gagcgccgga ggggcaaggg gcacgatggc ctttaccagg gtctcagtac agccaccaag 1500

gacacctacg acgcccttca catgcaggcc ctgccccctc gcggctctgg cgagggaagg 1560

ggttccctgc ttacttgcgg cgacgtcgaa gagaatcccg gtccgatggc cctcccagta 1620

actgccctcc ttttgcccct cgcactcctt cttcatgccg ctcgccccaa ctgggtcaac 1680

gtgattagcg atttgaagaa aatcgaggac cttatacagt ctatgcatat tgacgctaca 1740

ctgtatactg agagtgatgt acacccgtcc tgtaaggtaa cggccatgaa atgctttctt 1800

ctggagctcc aggtcatcag cttggagtct ggggacgcaa gcatccacga tacggttgaa 1860

aacctcatca tccttgcgaa caactctctc tcatctaatg gaaacgttac agagagtggg 1920

tgtaaggagt gcgaagagtt ggaagaaaaa aacatcaaag aatttcttca atccttcgtt 1980

cacatagtgc aaatgttcat taacacgtcc actaccacac ccgccccgag gccacctacg 2040

ccggcaccga ctatcgccag tcaacccctc tctctgcgcc ccgaggcttg ccggcctgcg 2100

gctggtgggg cggtccacac ccggggcctg gattttgcgt gcgatatata catctgggca 2160

cctcttgccg gcacctgcgg agtgctgctt ctctcactcg ttattacgct gtactgctaa 2220

gcggccgcgt cgac 2234

<210> 80

<211> 732

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-11b protein

<400> 80

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Met Asp

340 345 350

Gln Gln Ala Ile Tyr Ala Glu Leu Asn Leu Pro Thr Asp Ser Gly Pro

355 360 365

Glu Ser Ser Ser Pro Ser Ser Leu Pro Arg Asp Val Cys Gln Gly Ser

370 375 380

Pro Trp His Gln Phe Ala Leu Lys Leu Ser Cys Arg Val Lys Phe Ser

385 390 395 400

Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr

405 410 415

Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys

420 425 430

Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn

435 440 445

Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu

450 455 460

Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly

465 470 475 480

His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr

485 490 495

Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Glu Gly

500 505 510

Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro

515 520 525

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

530 535 540

His Ala Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys

545 550 555 560

Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr

565 570 575

Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe

580 585 590

Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile

595 600 605

His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser

610 615 620

Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu

625 630 635 640

Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val

645 650 655

Gln Met Phe Ile Asn Thr Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

660 665 670

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

675 680 685

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

690 695 700

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

705 710 715 720

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

725 730

<210> 81

<211> 2165

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-12b DNA

<400> 81

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actgtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg catgatcgaa 1080

acatacaacc aaacttctcc ccgatctgcg gccactggac tgcccatcag catgaaaaga 1140

gtgaagttca gcaggagcgc agacgccccc gcgtaccagc agggccagaa ccagctctat 1200

aacgagctca atctaggacg aagagaggag tacgatgttt tggacaagag acgtggccgg 1260

gaccctgaga tggggggaaa gccgagaagg aagaaccctc aggaaggcct gtacaatgaa 1320

ctgcagaaag ataagatggc ggaggcctac agtgagattg ggatgaaagg cgagcgccgg 1380

aggggcaagg ggcacgatgg cctttaccag ggtctcagta cagccaccaa ggacacctac 1440

gacgcccttc acatgcaggc cctgccccct cgcggctctg gcgagggaag gggttccctg 1500

cttacttgcg gcgacgtcga agagaatccc ggtccgatgg ccctcccagt aactgccctc 1560

cttttgcccc tcgcactcct tcttcatgcc gctcgcccca actgggtcaa cgtgattagc 1620

gatttgaaga aaatcgagga ccttatacag tctatgcata ttgacgctac actgtatact 1680

gagagtgatg tacacccgtc ctgtaaggta acggccatga aatgctttct tctggagctc 1740

caggtcatca gcttggagtc tggggacgca agcatccacg atacggttga aaacctcatc 1800

atccttgcga acaactctct ctcatctaat ggaaacgtta cagagagtgg gtgtaaggag 1860

tgcgaagagt tggaagaaaa aaacatcaaa gaatttcttc aatccttcgt tcacatagtg 1920

caaatgttca ttaacacgtc cactaccaca cccgccccga ggccacctac gccggcaccg 1980

actatcgcca gtcaacccct ctctctgcgc cccgaggctt gccggcctgc ggctggtggg 2040

gcggtccaca cccggggcct ggattttgcg tgcgatatat acatctgggc acctcttgcc 2100

ggcacctgcg gagtgctgct tctctcactc gttattacgc tgtactgcta agcggccgcg 2160

tcgac 2165

<210> 82

<211> 709

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-12b protein

<400> 82

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Met Ile

340 345 350

Glu Thr Tyr Asn Gln Thr Ser Pro Arg Ser Ala Ala Thr Gly Leu Pro

355 360 365

Ile Ser Met Lys Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala

370 375 380

Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg

385 390 395 400

Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu

405 410 415

Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn

420 425 430

Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met

435 440 445

Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly

450 455 460

Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala

465 470 475 480

Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys

485 490 495

Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala

500 505 510

Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp

515 520 525

Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser

530 535 540

Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser

545 550 555 560

Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile

565 570 575

Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu

580 585 590

Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu

595 600 605

Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu

610 615 620

Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser

625 630 635 640

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

645 650 655

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

660 665 670

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile

675 680 685

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

690 695 700

Ile Thr Leu Tyr Cys

705

<210> 83

<211> 2315

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-13b DNA

<400> 83

ggatccgaat tcgccgccac catggcctta ccagtgaccg ccttgctcct gccgctggcc 60

ttgctgctcc acgccgccag gccggactac aaagacgatg acgataaagg cggtggtggc 120

tctggtggtg gcggcagcga catccagatg acacagacta catcctccct gtctgcctct 180

ctgggagaca gagtcaccat cagttgcagg gcaagtcagg acattagtaa atatttaaat 240

tggtatcagc agaaaccaga tggaactgtt aaactcctga tctaccatac atcaagatta 300

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta ttctctcacc 360

attagcaacc tggagcaaga agatattgcc acttactttt gccaacaggg taatacgctt 420

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 480

ggtgggtcgg gtggcggcgg atctgaggtg aaactgcagg agtcaggacc tggcctggtg 540

gcgccctcac agagcctgtc cgtcacatgc actngtctcag gggtctcatt acccgactat 600

ggtgtaagct ggattcgcca gcctccacga aagggtctgg agtggctggg agtaatatgg 660

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat catcaaggac 720

aactccaaga gccaagtttt cttaaaaatg aacagtctgc aaactgatga cacagccatt 780

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 840

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 900

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 960

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg caacagtcga 1080

agaaggtgtg ggcagaagaa aaagctagtg atcaacagtg gcaatggagc tgtggaggac 1140

agaaagccaa gtggactcaa cggagaggcc agcaagtctc aggaaatggt gcatttggtg 1200

aacaaggagt cgtcagaaac tccagaccag tttatgacag ctgatgagac aaggaacctg 1260

cagaatgtgg acatgaagat tggggtgaga gtgaagttca gcaggagcgc agacgccccc 1320

gcgtaccagc agggccagaa ccagctctat aacgagctca atctaggacg aagagaggag 1380

tacgatgttt tggacaagag acgtggccgg gaccctgaga tggggggaaa gccgagaagg 1440

aagaaccctc aggaaggcct gtacaatgaa ctgcagaaag ataagatggc ggaggcctac 1500

agtgagattg ggatgaaagg cgagcgccgg aggggcaagg ggcacgatgg cctttaccag 1560

ggtctcagta cagccaccaa ggacacctac gacgcccttc acatgcaggc cctgccccct 1620

cgcggctctg gcgagggaag gggttccctg cttacttgcg gcgacgtcga agagaatccc 1680

ggtccgatgg ccctcccagt aactgccctc cttttgcccc tcgcactcct tcttcatgcc 1740

gctcgcccca actgggtcaa cgtgattagc gatttgaaga aaatcgagga ccttatacag 1800

tctatgcata ttgacgctac actgtatact gagagtgatg tacacccgtc ctgtaaggta 1860

acggccatga aatgctttct tctggagctc caggtcatca gcttggagtc tggggacgca 1920

agcatccacg atacggttga aaacctcatc atccttgcga acaactctct ctcatctaat 1980

ggaaacgtta cagagagtgg gtgtaaggag tgcgaagagt tggaagaaaa aaacatcaaa 2040

gaatttcttc aatccttcgt tcacatagtg caaatgttca ttaacacgtc cactaccaca 2100

cccgccccga ggccacctac gccggcaccg actatcgcca gtcaacccct ctctctgcgc 2160

cccgaggctt gccggcctgc ggctggtggg gcggtccaca cccggggcct ggattttgcg 2220

tgcgatatat acatctgggc acctcttgcc ggcacctgcg gagtgctgct tctctcactc 2280

gttattacgc tgtactgcta agcggccgcg tcgac 2315

<210> 84

<211> 759

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19-13b protein

<400> 84

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser

35 40 45

Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu

100 105 110

Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser

165 170 175

Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe

225 230 235 240

Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn Ser

340 345 350

Arg Arg Arg Cys Gly Gln Lys Lys Lys Leu Val Ile Asn Ser Gly Asn

355 360 365

Gly Ala Val Glu Asp Arg Lys Pro Ser Gly Leu Asn Gly Glu Ala Ser

370 375 380

Lys Ser Gln Glu Met Val His Leu Val Asn Lys Glu Ser Ser Glu Thr

385 390 395 400

Pro Asp Gln Phe Met Thr Ala Asp Glu Thr Arg Asn Leu Gln Asn Val

405 410 415

Asp Met Lys Ile Gly Val Arg Val Lys Phe Ser Arg Ser Ala Asp Ala

420 425 430

Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu

435 440 445

Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp

450 455 460

Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu

465 470 475 480

Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile

485 490 495

Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr

500 505 510

Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met

515 520 525

Gln Ala Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu

530 535 540

Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val

545 550 555 560

Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro

565 570 575

Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile

580 585 590

Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His

595 600 605

Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln

610 615 620

Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu

625 630 635 640

Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val

645 650 655

Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile

660 665 670

Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn

675 680 685

Thr Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr

690 695 700

Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala

705 710 715 720

Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile

725 730 735

Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser

740 745 750

Leu Val Ile Thr Leu Tyr Cys

755

<210> 85

<211> 6488

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19 DNA

<400> 85

atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca 60

tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc 120

agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag 180

aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt 240

tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 300

cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 360

cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat 420

aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc 480

agattgattg actgcccacc tcggggggtct ttcatttgga ggttccaccg agatttggag 540

acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc 600

gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg 660

tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga 720

acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg 780

acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga 840

gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga 900

agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact 960

gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc 1020

ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag 1080

agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga 1140

gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc 1200

ccgcatggac acccagacca ggtccctac atcgtgacct gggaagcctt ggcttttgac 1260

ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc 1320

gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca 1380

gccctcactc cttctctagg cgccggaatt cgttaacctc gagcgggatc aattccgccc 1440

cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat 1500

gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt 1560

cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt 1620

gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc 1680

gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc 1740

acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat 1800

agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc 1860

ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg 1920

tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct 1980

ttgaaaaaca cgataatacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 2040

ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 2100

gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 2160

tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 2220

gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 2280

tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 2340

agttcgagg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 2400

acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 2460

tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 2520

acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 2580

tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 2640

agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 2700

tggacgagct gtacaagtaa agcggccgcg actctagagt cgacctgcag gcatgcaagc 2760

ttcaggtagc cggctaacgt taacaaccgg tacctctaga actatagcta gcatgcgcaa 2820

atttaaagcg ctgatatcga taaaataaaa gattttattt agtctccaga aaaagggggg 2880

aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc 2940

atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag 3000

cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcaggggccaa 3060

gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt 3120

ttccaagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 3180

tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 3240

ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa 3300

taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc 3360

tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga 3420

gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt 3480

tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc 3540

tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 3600

ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 3660

tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 3720

cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 3780

ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 3840

ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 3900

gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 3960

gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4020

taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4080

accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4140

tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4200

cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4260

agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 4320

gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 4380

gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 4440

tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 4500

acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 4560

cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 4620

acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 4680

acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 4740

tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 4800

ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 4860

ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 4920

tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 4980

aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5040

ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5100

ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5160

gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 5220

gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 5280

cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 5340

actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 5400

ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 5460

tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 5520

ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 5580

agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 5640

aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc 5700

attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg 5760

cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct 5820

tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 5880

gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 5940

atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg 6000

ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 6060

cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc 6120

cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca 6180

acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc 6240

cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg 6300

cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat 6360

ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc 6420

tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa 6480

agggggga 6488

<210> 86

<211> 486

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> NK19 protein

<400> 86

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu

20 25 30

Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile

85 90 95

Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu

130 135 140

Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln Ser

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu Lys

210 215 220

Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg

325 330 335

Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln

340 345 350

Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu

355 360 365

Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala

370 375 380

Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu

385 390 395 400

Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp

405 410 415

Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu

420 425 430

Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile

435 440 445

Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr

450 455 460

Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met

465 470 475 480

Gln Ala Leu Pro Pro Arg

485

<210> 87

<211> 6488

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> MSCV-IRES-GFP expression plasmid

<400> 87

atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca 60

tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc 120

agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag 180

aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt 240

tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 300

cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 360

cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat 420

aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc 480

agattgattg actgcccacc tcggggggtct ttcatttgga ggttccaccg agatttggag 540

acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc 600

gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg 660

tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga 720

acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg 780

acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga 840

gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga 900

agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact 960

gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc 1020

ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag 1080

agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga 1140

gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc 1200

ccgcatggac acccagacca ggtccctac atcgtgacct gggaagcctt ggcttttgac 1260

ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc 1320

gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca 1380

gccctcactc cttctctagg cgccggaatt cgttaacctc gagcgggatc aattccgccc 1440

cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat 1500

gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt 1560

cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt 1620

gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc 1680

gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc 1740

acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat 1800

agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc 1860

ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg 1920

tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct 1980

ttgaaaaaca cgataatacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 2040

ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 2100

gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 2160

tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 2220

gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 2280

tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 2340

agttcgagg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 2400

acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 2460

tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 2520

acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 2580

tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 2640

agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 2700

tggacgagct gtacaagtaa agcggccgcg actctagagt cgacctgcag gcatgcaagc 2760

ttcaggtagc cggctaacgt taacaaccgg tacctctaga actatagcta gcatgcgcaa 2820

atttaaagcg ctgatatcga taaaataaaa gattttattt agtctccaga aaaagggggg 2880

aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc 2940

atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag 3000

cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcaggggccaa 3060

gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt 3120

ttccaagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 3180

tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 3240

ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa 3300

taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc 3360

tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga 3420

gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt 3480

tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc 3540

tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 3600

ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 3660

tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 3720

cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 3780

ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 3840

ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 3900

gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 3960

gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4020

taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4080

accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4140

tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4200

cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4260

agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 4320

gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 4380

gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 4440

tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 4500

acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 4560

cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 4620

acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 4680

acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 4740

tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 4800

ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 4860

ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 4920

tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 4980

aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5040

ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5100

ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5160

gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 5220

gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 5280

cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 5340

actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 5400

ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 5460

tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 5520

ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 5580

agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 5640

aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc 5700

attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg 5760

cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct 5820

tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 5880

gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 5940

atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg 6000

ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 6060

cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc 6120

cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca 6180

acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc 6240

cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg 6300

cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat 6360

ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc 6420

tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa 6480

agggggga 6488

<210> 88

<211> 117

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> heavy chain to CLDN6

<400> 88

Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr

20 25 30

Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile

35 40 45

Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Ile Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Gly Phe Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser

115

<210> 89

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> light chain 1 to CLDN6

<400> 89

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Leu

20 25 30

His Trp Phe Gln Gln Lys Pro Gly Thr Ser Pro Lys Leu Trp Val Tyr

35 40 45

Ser Thr Ser Asn Leu Pro Ser Gly Val Pro Ala Arg Phe Gly Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Ile Tyr Pro Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 90

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> light chain 2 to CLDN6

<400> 90

Gln Ile Val Leu Thr Gln Ser Pro Ser Ile Met Ser Val Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

His Trp Phe Gln Gln Lys Pro Gly Thr Ser Pro Lys Leu Gly Ile Tyr

35 40 45

Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Arg

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Ala Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Asn Tyr Pro Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 91

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> light chain 3 to CLDN6

<400> 91

Gln Ile Val Leu Thr Gln Ser Pro Ser Ile Met Ser Val Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

His Trp Phe Gln Gln Lys Pro Gly Thr Ser Pro Lys Leu Ser Ile Tyr

35 40 45

Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Arg

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Ala Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Asn Tyr Pro Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 92

<211> 8

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain CDR1 to CLDN6

<400> 92

Gly Tyr Ser Phe Thr Gly Tyr Thr

1 5

<210> 93

<211> 8

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain CDR2 to CLDN6

<400> 93

Ile Asn Pro Tyr Asn Gly Gly Thr

1 5

<210> 94

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Heavy chain CDR3 to CLDN6

<400> 94

Ala Arg Asp Tyr Gly Phe Val Leu Asp Tyr

1 5 10

<210> 95

<211> 5

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 light chain 1 to CLDN6

<400> 95

Ser Ser Val Ser Tyr

1 5

<210> 96

<211> 3

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 light chain 1 to CLDN6

<400> 96

Ser Thr Ser

1

<210> 97

<211> 8

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 light chain 1 to CLDN6

<400> 97

Gln Gln Arg Ser Ile Tyr Pro Pro

1 5

<210> 98

<211> 5

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 light chain 2 to CLDN6

<400> 98

Ser Ser Val Ser Tyr

1 5

<210> 99

<211> 3

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 light chain 2 to CLDN6

<400> 99

Ser Thr Ser

1

<210> 100

<211> 8

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 light chain 2 to CLDN6

<400> 100

Gln Gln Arg Ser Asn Tyr Pro Pro

1 5

<210> 101

<211> 5

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 LC to CLDN6

<400> 101

Ser Ser Val Ser Tyr

1 5

<210> 102

<211> 3

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 LC3 to CLDN6

<400> 102

Ser Thr Ser

1

<210> 103

<211> 8

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 LC3 to CLDN6

<400> 103

Gln Gln Arg Ser Asn Tyr Pro Pro

1 5

<210> 104

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CD19 heavy chain variable region

<400> 104

Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 105

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> variable region of light chain to CD19

<400> 105

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Ile Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Ala Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 106

<211> 450

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CD19 heavy chain variable region

<400> 106

Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Lys

450

<210> 107

<211> 214

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> variable region of light chain to CD19

<400> 107

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Ile Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Ala Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 108

<211> 11

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 LC to CD19

<400> 108

Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn

1 5 10

<210> 109

<211> 7

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 LC to CD19

<400> 109

His Thr Ser Arg Leu His Ser

1 5

<210> 110

<211> 9

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 LC to CD19

<400> 110

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

1 5

<210> 111

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HC to CD19

<400> 111

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 112

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HC to CD19

<400> 112

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Ser Ser Leu Lys Ser

1 5 10 15

<210> 113

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HC to CD19

<400> 113

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Gln Ser Ser Leu Lys Ser

1 5 10 15

<210> 114

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HC to CD19

<400> 114

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ser Leu Lys Ser

1 5 10 15

<210> 115

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HC to CD19

<400> 115

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 116

<211> 242

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CAR to CD19

<400> 116

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu

115 120 125

Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys

130 135 140

Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg

145 150 155 160

Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Val Ile Trp Gly Ser

165 170 175

Glu Thr Thr Tyr Tyr Gln Ser Ser Leu Lys Ser Arg Val Thr Ile Ser

180 185 190

Lys Asp Asn Ser Lys Asn Gln Val Ser Leu Lys Leu Ser Ser Val Thr

195 200 205

Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly

210 215 220

Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

225 230 235 240

Ser Ser

<210> 117

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> huFMC63VLv1

<400> 117

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

100 105

<210> 118

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223>HUFMC63VLV2

<400> 118

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

100 105

<210> 119

<211> 107

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223>HUFMC63VLV3

<400> 119

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

100 105

<210> 120

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223>HUFMC63VHV1

<400> 120

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 121

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223>HUFMC63VHV2

<400> 121

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 122

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223>HUFMC63VHV3

<400> 122

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 123

<211> 120

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223>HUFMC63VHV4

<400> 123

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu

65 70 75 80

Lys Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 124

<211> 11

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 huFMC63VLv1

<400> 124

Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn

1 5 10

<210> 125

<211> 7

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 huFMC63VLv1

<400> 125

His Thr Ser Arg Leu His Ser

1 5

<210> 126

<400> 126

000

<210> 127

<211> 11

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HUFMC63VLV2

<400> 127

Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn

1 5 10

<210> 128

<211> 7

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HUFMC63VLV2

<400> 128

His Thr Ser Arg Leu His Ser

1 5

<210> 129

<211> 9

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 HUFMC63VLV2

<400> 129

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

1 5

<210> 130

<211> 11

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HUFMC63VLV3

<400> 130

Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn

1 5 10

<210> 131

<211> 7

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HUFMC63VLV3

<400> 131

His Thr Ser Arg Leu His Ser

1 5

<210> 132

<211> 9

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 HUFMC63VLV3

<400> 132

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

1 5

<210> 133

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HUFMC63VHV1

<400> 133

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 134

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HUFMC63VHV1

<400> 134

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

1 5 10 15

<210> 135

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 HUFMC63VHV1

<400> 135

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 136

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HUFMC63VHV2

<400> 136

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 137

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HUFMC63VHV2

<400> 137

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

1 5 10 15

<210> 138

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 HUFMC63VHV2

<400> 138

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 139

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HUFMC63VHV3

<400> 139

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 140

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HUFMC63VHV3

<400> 140

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

1 5 10 15

<210> 141

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 HUFMC63VHV3

<400> 141

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 142

<211> 10

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR1 HUFMC63VHV4

<400> 142

Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

1 5 10

<210> 143

<211> 16

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR2 HUFMC63VHV4

<400> 143

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

1 5 10 15

<210> 144

<211> 12

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> CDR3 HUFMC63VHV4

<400> 144

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 145

<211> 1122

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NKG2D-Ox40-CD3z

<400> 145

atggccttac cagtgaccgc cttgctcctg ccgctggcct tgctgctcca cgccgccagg 60

ccgttattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 120

aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 180

tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 240

gaggaccagg atttacttaa actggtgaag tcatatcatt ggatgggact agtacacatt 300

ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 360

ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggctatata 420

gaaaactgtt caactccaaa tacgtacatc tgcatgcaaa ggactgtgac cacgacgcca 480

gcgccgcgac caccaacacc ggcgcccacc atcgcgtcgc agcccctgtc cctgcgccca 540

gaggcgtgcc ggccagcggc ggggggcgca gtgcacacga gggggctgga cttcgcctgt 600

gatatctaca tctgggcgcc cttggccggg acttgtgggg tccttctcct gtcactggtt 660

atcacccttt actgccggag ggaccagagg ctgccccccg atgcccacaa gccccctggg 720

ggaggcagtt tccggacccc catccaagag gagcaggccg acgcccactc caccctggcc 780

aagatcagag tgaagttcag caggagcgca gacgcccccg cgtaccagca gggccagaac 840

cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga 900

cgtggccggg accctgagat ggggggaaag ccgagaagga agaaccctca ggaaggcctg 960

tacaatgaac tgcagaaaga taagatggcg gaggcctaca gtgagattgg gatgaaaggc 1020

gagcgccgga ggggcaaggg gcacgatggc ctttaccagg gtctcagtac agccaccaag 1080

gacacctacg acgcccttca catgcaggcc ctgccccctc gc 1122

<210> 146

<211> 341

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NKG2D-OX40-CD3z

<400> 146

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr

20 25 30

Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn

35 40 45

Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln

50 55 60

Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys

65 70 75 80

Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly

85 90 95

Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser

100 105 110

Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp

115 120 125

Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser

130 135 140

Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val Glu Ser Lys Tyr

145 150 155 160

Gly Pro Pro Cys Pro Ser Cys Pro Ile Tyr Ile Trp Ala Pro Leu Ala

165 170 175

Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys

180 185 190

Arg Arg Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly

195 200 205

Gly Ser Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser

210 215 220

Thr Leu Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro

225 230 235 240

Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly

245 250 255

Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro

260 265 270

Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr

275 280 285

Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly

290 295 300

Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln

305 310 315 320

Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln

325 330 335

Ala Leu Pro Pro Arg

340

<210> 147

<211> 1467

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA sequence FMC63_CD28_CAR

<400> 147

atgcttctcc tggtgacaag ccttctttac cacaccagc attcctcctg atcccagaca 60

tccagataca tcctccctgt ctgcctctct gggagacaga gtcaccgggc aagtcaggac 120

attagtaaat atttaaattg gtatcagctc tgtgagtgac acagacatca gttgcagcag 180

aaaccagatg gaactgttaa actcctgatc taccatacat caagattaca ctcaggagtc 240

ccatcaaggt tcagtggcag tgggtctgga acagattatt ctctcaccat tagcaacctg 300

gagcaagaag atattgccac ttacttttgc caacagggta atacgcttcc gtacacgttc 360

ggagggggga ctaagttgga aataacaggc tccacctctg gatccggcaa gcccggatct 420

ggcgagggat ccaccaaggg cgaggtgaaa ctgcaggagt caggacctgg cctggtggcg 480

ccctcacaga gcctgtccgt cacatgcact gtctcagggg tctcattacc cgactatggt 540

gtaagctgga ttcgccagcc tccacgaaag ggtctggagt ggctgggagt aatatggggt 600

agtgaaacca catactataa ttcagctctc aaatccagac tgaccatcat caaggacaac 660

tccaagagcc aagttttctt aaaaatgaac agtctgcaaa ctgatgacac agccatttac 720

tactgtgcca aacattatta ctacggtggt agctatgcta tggactactg gggtcaagga 780

acctcagtca ccgtctcctc agcggccgca attgaagtta tgtatcctcc tccttaccta 840

gacaatgaga agagcaatgg aaccattatc catgtgaaag ggaaacacct ttgtccaagt 900

cccctatttc ccggaccttc taagcccttt tgggtgctgg tggtggttgg gggagtcctg 960

gcttgctata gcttgctagt aacagtggcc tttattattt tctgggtgag gagtaagagg 1020

agcaggctcc tgcacagtga ctacatgaac atgactcccc gccgccccgg gcccacccgc 1080

aagcattacc agccctatgc cccaccacgc gacttcgcag cctatcgctc cagagtgaag 1140

ttcagcagga gcgcagacgc ccccgcgtac cagcagggcc agaaccagct ctataacgag 1200

ctcaatctag gacgaagaga ggagtacgat gttttggaca agagacgtgg ccgggaccct 1260

gagatggggg gaaagccgag aaggaagaac cctcaggaag gcctgtacaa tgaactgcag 1320

aaagataaga tggcggaggc ctacagtgag attgggatga aaggcgagcg ccggaggggc 1380

aaggggcacg atggccttta ccagggtctc agtacagcca ccaaggacac ctacgacgcc 1440

cttcacatgc aggccctgcc ccctcgc 1467

<210> 148

<211> 331

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> FCM63 light chain DNA sequence

<400> 148

gacatccaga tacatcctcc ctgtctgcct ctctgggaga cagagtcacc gggcaagtca 60

ggacattagt aaatatttaa attggtatca gctctgtgag tgacacagac atcagttgca 120

gcagaaacca gatggaactg ttaaactcct gatctaccat acatcaagat tacactcagg 180

agtcccatca aggttcagtg gcagtgggtc tggaacagat tattctctca ccattagcaa 240

cctggagcaa gaagatattg ccacttactt ttgccaacag ggtaatacgc ttccgtacac 300

gttcggaggg gggactaagt tggaaataac a 331

<210> 149

<211> 54

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> FMC63 linker DNA sequence

<400> 149

ggctccacct ctggatccgg caagcccgga tctggcgagg gatccaccaa gggc 54

<210> 150

<211> 360

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> FMC63 heavy chain DNA sequence

<400> 150

gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60

acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120

ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180

tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240

aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300

tacggtggta gctatgctat ggactactgg ggtcaaggaa cctcagtcac cgtctcctca 360

<210> 151

<211> 117

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> CD28 spacer DNA sequence

<400> 151

attgaagtta tgtatcctcc tccttaccta gacaatgaga agagcaatgg aaccattatc 60

catgtgaaag ggaaacacct ttgtccaagt cccctatttc ccggaccttc taagccc 117

<210> 152

<211> 81

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA sequence of the transmembrane region

<400> 152

ttttgggtgc tggtggtggt tgggggagtc ctggcttgct atagcttgct agtaacagtg 60

gcctttatta ttttctgggt g 81

<210> 153

<211> 123

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> CD28 costimulatory domain DNA sequence

<400> 153

aggagtaaga ggagcaggct cctgcacagt gactacatga acatgactcc ccgccgcccc 60

gggcccaccc gcaagcatta ccagccctat gccccaccac gcgacttcgc agcctatcgc 120

tcc 123

<210> 154

<211> 336

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> CD3-zeta [repeat]

<400> 154

agagtgaagt tcagcaggag cgcagacgcc cccgcgtacc agcagggcca gaaccagctc 60

tataacgagc tcaatctagg acgaagagag gagtacgatg ttttggacaa gagacgtggc 120

cgggaccctg agatgggggg aaagccgaga aggaagaacc ctcaggaagg cctgtacaat 180

gaactgcaga aagataagat ggcggaggcc tacagtgaga ttgggatgaa aggcgagcgc 240

cggaggggca aggggcacga tggcctttac cagggtctca gtacagccac caaggacacc 300

tacgacgccc ttcacatgca ggccctgccc cctcgc 336

<210> 155

<211> 342

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> mbIL15 coding sequence

<400> 155

aactgggtca acgtgattag cgatttgaag aaaatcgagg accttataca gtctatgcat 60

attgacgcta cactgtatac tgagagtgat gtacacccgt cctgtaaggt aacggccatg 120

aaatgctttc ttctggagct ccaggtcatc agcttggagt ctggggacgc aagcatccac 180

gatacggttg aaaacctcat catccttgcg aacaactctc tctcatctaa tggaaacgtt 240

acagagagtg ggtgtaagga gtgcgaagag ttggaagaaa aaaacatcaa agaatttctt 300

caatccttcg ttcacatagt gcaaatgttc attaacacgt cc 342

<210> 156

<211> 135

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> Hinge DNA sequence cd, reverse transcription c#: 157

<400> 156

accaccaccc cggcgccgcg cccgccgacc ccggcgccga ccattgcgag ccagccgctg 60

agcctgcgcc cggaagcgtg ccgcccggcg gcgggcggcg cggtgcatac ccgcggcctg 120

gattttgcgt gcgat 135

<210> 157

<211> 45

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> Amino acid sequence of CD8 hinge

<400> 157

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

1 5 10 15

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

20 25 30

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

35 40 45

<210> 158

<211> 72

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> CD8 TM DNA sequence, reverse transcription from No: 159

<400> 158

atttatattt gggcgccgct ggcgggcacc tgcggcgtgc tgctgctgag cctggtgatt 60

accctgtatt gc 72

<210> 159

<211> 24

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of TM CD8

<400> 159

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

1 5 10 15

Ser Leu Val Ile Thr Leu Tyr Cys

20

<210> 160

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-1 DNA

<400> 160

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cgggcggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attttaccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat tattgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcctgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gggcaaaggc ctggaatgga ttggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaaaccagg tgagcctgaa actgagcagc gtgaccgcgg cggataccgc ggtgtattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 161

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-1

<400> 161

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Ile Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 162

<211> 2187

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-2 DNA

<400> 162

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cgggcggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attttaccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat ttttgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcctgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gggcaaaggc ctggaatgga ttggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaaaccagg tgagcctgaa actgagcagc gtgaccgcgg cggataccgc ggtgtattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgc 2187

<210> 163

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-2

<400> 163

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Ile Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 164

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-3 DNA

<400> 164

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cggatggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attataccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat ttttgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcctgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gggcaaaggc ctggaatgga ttggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaaaccagg tgagcctgaa actgagcagc gtgaccgcgg cggataccgc ggtgtattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 165

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-3

<400> 165

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Ile Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 166

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-4

<400> 166

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cgggcggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attttaccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat tattgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcctgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gggcaaaggc ctggaatggc tgggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaagccagg tgagcctgaa actgagcagc gtgaccgcgg cggataccgc ggtgtattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 167

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-4

<400> 167

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 168

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-5 DNA

<400> 168

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cgggcggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attttaccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat ttttgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcctgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gggcaaaggc ctggaatggc tgggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaagccagg tgagcctgaa actgagcagc gtgaccgcgg cggataccgc ggtgtattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 169

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-5

<400> 169

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 170

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-6

<400> 170

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgacatcca gatgacacag agcccgtcct ccctgtctgc ctctgtggga 180

gacagagtca ccatcacctg cagggcaagt caggacatta gtaaatattt aaattggtat 240

cagcagaaac cagacggaac tgttaaactc ctgatctacc atacatcaag attacactca 300

ggagtcccat caaggttcag tggcagtggg tctggaacag attacaccct caccattagc 360

agcctgcaac cggaagatat tgccacttac ttctgccaac agggtaatac gcttccgtac 420

acgttcggag gggggaccaa gctggagatc acaggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggagtcag gacctggcct ggtgaaaccc 540

tcacagactc tgtccctgac atgcactgtc tcaggggtct cattacccga ctatggtgta 600

agctggattc gccagcctcc aggtaagggt ctggagtggc tgggagtaat atggggtagt 660

gaaaccacat actataattc agctctcaaa tccagactga ccatctccaa ggacaactcc 720

aagagccaag tttccttaaa attaagtagt gttactgctg ctgacacagc cgtctactac 780

tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg ccaaggaacc 840

tcagtcaccg tctcctcaac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 171

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-6

<400> 171

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 172

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-7

<400> 172

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgacatcca gatgacacag agcccgtcct ccctgtctgc ctctgtggga 180

gacagagtca ccatcacctg cagggcaagt caggacatta gtaaatattt aaattggtat 240

cagcagaaac caggtggaac tgttaaactc ctgatctacc atacatcaag attacactca 300

ggagtcccat caaggttcag tggcagtggg tctggaacag atttcaccct caccattagc 360

agcctgcaac cggaagatat tgccacttac tactgccaac agggtaatac gcttccgtac 420

acgttcggag gggggaccaa gctggagatc acaggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggagtcag gacctggcct ggtgaaaccc 540

tcacagactc tgtccgtgac atgcactgtc tcaggggtct cattacccga ctatggtgta 600

agctggattc gccagcctcc aggtaagggt ctggagtggc tgggagtaat atggggtagt 660

gaaaccacat actataattc agctctcaaa tccagactga ccatctccaa ggacaactcc 720

aagagccaag tttccttaaa attaagtagt gttactgctg ctgacacagc cgtctactac 780

tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg ccaaggaacc 840

tcagtcaccg tctcctcaac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 173

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-7

<400> 173

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 174

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-8

<400> 174

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgacatcca gatgacacag agcccgtcct ccctgtctgc ctctgtggga 180

gacagagtca ccatcacctg cagggcaagt caggacatta gtaaatattt aaattggtat 240

cagcagaaac caggtggaac tgttaaactc ctgatctacc atacatcaag attacactca 300

ggagtcccat caaggttcag tggcagtggg tctggaacag atttcaccct caccattagc 360

agcctgcaac cggaagatat tgccacttac ttctgccaac agggtaatac gcttccgtac 420

acgttcggag gggggaccaa gctggagatc acaggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggagtcag gacctggcct ggtgaaaccc 540

tcacagactc tgtccgtgac atgcactgtc tcaggggtct cattacccga ctatggtgta 600

agctggattc gccagcctcc aggtaagggt ctggagtggc tgggagtaat atggggtagt 660

gaaaccacat actataattc agctctcaaa tccagactga ccatctccaa ggacaactcc 720

aagagccaag tttccttaaa attaagtagt gttactgctg ctgacacagc cgtctactac 780

tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg ccaaggaacc 840

tcagtcaccg tctcctcaac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 175

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-8

<400> 175

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 176

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-9

<400> 176

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgacatcca gatgacacag agcccgtcct ccctgtctgc ctctgtggga 180

gacagagtca ccatcacctg cagggcaagt caggacatta gtaaatattt aaattggtat 240

cagcagaaac cagacggaac tgttaaactc ctgatctacc atacatcaag attacactca 300

ggagtcccat caaggttcag tggcagtggg tctggaacag attacaccct caccattagc 360

agcctgcaac cggaagatat tgccacttac ttctgccaac agggtaatac gcttccgtac 420

acgttcggag gggggaccaa gctggagatc acaggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggagtcag gacctggcct ggtgaaaccc 540

tcacagactc tgtccgtgac atgcactgtc tcaggggtct cattacccga ctatggtgta 600

agctggattc gccagcctcc aggtaagggt ctggagtggc tgggagtaat atggggtagt 660

gaaaccacat actataattc agctctcaaa tccagactga ccatctccaa ggacaactcc 720

aagagccaag tttccttaaa attaagtagt gttactgctg ctgacacagc cgtctactac 780

tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg ccaaggaacc 840

tcagtcaccg tctcctcaac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 177

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-9

<400> 177

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 178

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-10

<400> 178

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cgggcggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attttaccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat tattgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcgtgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gcgcaaaggc ctggaatggc tgggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaagccagg tgagcctgaa aatgagcagc gtgaccgcgg cggataccgc gatttattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 179

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-10

<400> 179

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 180

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-11

<400> 180

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cgggcggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attttaccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat ttttgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcgtgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gcgcaaaggc ctggaatggc tgggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaagccagg tgagcctgaa aatgagcagc gtgaccgcgg cggataccgc gatttattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 181

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-11

<400> 181

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 182

<211> 2204

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-12

<400> 182

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga ctacaaagac gatgacgata aaggcggtgg tggctctggt 120

ggtggcggca gcgatattca gatgacccag agcccgagca gcctgagcgc gagcgtgggc 180

gatcgcgtga ccattacctg ccgcgcgagc caggatatta gcaaatatct gaactggtat 240

cagcagaaac cggatggcac cgtgaaactg ctgatttatc ataccagccg cctgcatagc 300

ggcgtgccga gccgctttag cggcagcggc agcggcaccg attataccct gaccattagc 360

agcctgcagc cggaagatat tgcgacctat ttttgccagc agggcaacac cctgccgtat 420

acctttggcg gcggcaccaa actggaaatt accggtggcg gtggctcggg cggtggtggg 480

tcgggtggcg gcggatctca ggtgcagctg caggaaagcg gcccgggcct ggtgaaaccg 540

agccagaccc tgagcgtgac ctgcaccgtg agcggcgtga gcctgccgga ttatggcgtg 600

agctggattc gccagccgcc gcgcaaaggc ctggaatggc tgggcgtgat ttggggcagc 660

gaaaccacct attataacag cgcgctgaaa agccgcctga ccattagcaa agataacagc 720

aaaagccagg tgagcctgaa aatgagcagc gtgaccgcgg cggataccgc gatttattat 780

tgcgcgaaac attattatta tggcggcagc tatgcgatgg attattgggg ccagggcacc 840

agcgtgaccg tgagcagcac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 900

atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 960

gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1020

acttgtgggg tccttctcct gtcactggtt atcacccttt actgccggag ggaccagagg 1080

ctgccccccg atgcccacaa gccccctggg ggaggcagtt tccggacccc catccaagag 1140

gagcaggccg acgcccactc caccctggcc aagatcagag tgaagttcag caggagcgca 1200

gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga 1260

agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag 1320

ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg 1380

gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc 1440

ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc 1500

ctgccccctc gcggctctgg cgagggaagg ggttccctgc ttacttgcgg cgacgtcgaa 1560

gagaatcccg gtccgatggc cctcccagta actgccctcc ttttgcccct cgcactcctt 1620

cttcatgccg ctcgccccaa ctgggtcaac gtgattagcg atttgaagaa aatcgaggac 1680

cttatacagt ctatgcatat tgacgctaca ctgtatactg agagtgatgt acacccgtcc 1740

tgtaaggtaa cggccatgaa atgctttctt ctggagctcc aggtcatcag cttggagtct 1800

ggggacgcaa gcatccacga tacggttgaa aacctcatca tccttgcgaa caactctctc 1860

tcatctaatg gaaacgttac agagagtggg tgtaagggagt gcgaagagtt ggaagaaaaa 1920

aacatcaaag aatttcttca atccttcgtt cacatagtgc aaatgttcat taacacgtcc 1980

actaccacac ccgccccgag gccacctacg ccggcaccga ctatcgccag tcaacccctc 2040

tctctgcgcc ccgaggcttg ccggcctgcg gctggtgggg cggtccacac ccggggcctg 2100

gattttgcgt gcgatatata catctgggca cctcttgccg gcacctgcgg agtgctgctt 2160

ctctcactcg ttattacgct gtactgctaa gcggccgcgt cgac 2204

<210> 183

<211> 724

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-12

<400> 183

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Tyr Lys Asp Asp Asp Asp Lys Gly Gly Gly

20 25 30

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

35 40 45

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

50 55 60

Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp

65 70 75 80

Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly

85 90 95

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu

100 105 110

Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln

115 120 125

Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu

130 135 140

Ile Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser

165 170 175

Gln Thr Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp

180 185 190

Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp

195 200 205

Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu

210 215 220

Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser

225 230 235 240

Leu Lys Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys

245 250 255

Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly

260 265 270

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

275 280 285

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

290 295 300

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

305 310 315 320

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

325 330 335

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg

340 345 350

Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser

355 360 365

Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu

370 375 380

Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

385 390 395 400

Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

405 410 415

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

420 425 430

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

435 440 445

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

450 455 460

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

465 470 475 480

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

485 490 495

Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly

500 505 510

Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu

515 520 525

Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val

530 535 540

Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met

545 550 555 560

His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys

565 570 575

Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser

580 585 590

Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile

595 600 605

Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser

610 615 620

Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe

625 630 635 640

Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr

645 650 655

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

660 665 670

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

675 680 685

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

690 695 700

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

705 710 715 720

Thr Leu Tyr Cys

<210> 184

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-NF-1 DNA

<400> 184

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccggg cggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgattt taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattatt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cctgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgggc aaaggcctgg aatggattgg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa accaggtgag cctgaaactg agcagcgtga ccgcggcgga taccgcggtg 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 185

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-1

<400> 185

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 186

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-NF-2 DNA

<400> 186

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccggg cggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgattt taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattttt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cctgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgggc aaaggcctgg aatggattgg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa accaggtgag cctgaaactg agcagcgtga ccgcggcgga taccgcggtg 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 187

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-2

<400> 187

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 188

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-NF-3 DNA

<400> 188

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccgga tggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgatta taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattttt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cctgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgggc aaaggcctgg aatggattgg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa accaggtgag cctgaaactg agcagcgtga ccgcggcgga taccgcggtg 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 189

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-3

<400> 189

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 190

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-NF-4 DNA

<400> 190

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccggg cggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgattt taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattatt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cctgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgggc aaaggcctgg aatggctggg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa gccaggtgag cctgaaactg agcagcgtga ccgcggcgga taccgcggtg 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 191

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-4

<400> 191

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 192

<211> 2153

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-NF-5 DNA

<400> 192

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccggg cggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgattt taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattttt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg agtcaggacc tggcctggtg 480

aaaccctcac agactctgtc cctgacatgc actgtctcag gggtctcatt acccgactat 540

ggtgtaagct ggattcgcca gcctccaggt aagggtctgg agtggctggg agtaatatgg 600

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat ctccaaggac 660

aactccaaga gccaagtttc cttaaaatta agtagtgtta ctgctgctga cacagccgtc 720

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 780

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcaga 1140

tctagcgcag acgcccccgc gtaccagcag ggccagaacc agctctataa cgagctcaat 1200

ctaggacgaa gagaggagta cgatgttttg gacaagagac gtggccggga ccctgagatg 1260

gggggaaagc cgagaaggaa gaaccctcag gaaggcctgt acaatgaact gcagaaagat 1320

aagatggcgg aggcctacag tgagattggg atgaaaggcg agcgccggag gggcaagggg 1380

cacgatggcc tttaccaggg tctcagtaca gccaccaagg acacctacga cgcccttcac 1440

atgcaggccc tgccccctcg cggctctggc gagggaaggg gttccctgct tacttgcggc 1500

gacgtcgaag agaatcccgg tccgatggcc ctcccagtaa ctgccctcct tttgcccctc 1560

gcactccttc ttcatgccgc tcgccccaac tgggtcaacg tgattagcga tttgaagaaa 1620

atcgaggacc ttatacagtc tatgcatatt gacgctacac tgtatactga gagtgatgta 1680

cacccgtcct gtaaggtaac ggccatgaaa tgctttcttc tggagctcca ggtcatcagc 1740

ttggagtctg gggacgcaag catccacgat acggttgaaa acctcatcat ccttgcgaac 1800

aactctctct catctaatgg aaacgttaca gagagtgggt gtaaggagtg cgaagagttg 1860

gaagaaaaaa acatcaaaga atttcttcaa tccttcgttc acatagtgca aatgttcatt 1920

aacacgtcca ctaccacacc cgccccgagg ccacctacgc cggcaccgac tatcgccagt 1980

caacccctct ctctgcgccc cgaggcttgc cggcctgcgg ctggtggggc ggtccacacc 2040

cggggcctgg attttgcgtg cgatatatac atctgggcac ctcttgccgg cacctgcgga 2100

gtgctgcttc tctcactcgt tattacgctg tactgctaag cggccgcgtc gac 2153

<210> 193

<211> 707

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-5

<400> 193

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ser Ala Asp Ala Pro Ala Tyr Gln

370 375 380

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

385 390 395 400

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

405 410 415

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

420 425 430

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

435 440 445

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

450 455 460

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

465 470 475 480

Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp

485 490 495

Val Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu

500 505 510

Leu Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn

515 520 525

Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His

530 535 540

Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys

545 550 555 560

Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu

565 570 575

Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile

580 585 590

Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly

595 600 605

Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu

610 615 620

Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr

625 630 635 640

Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln

645 650 655

Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala

660 665 670

Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala

675 680 685

Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr

690 695 700

Leu Tyr Cys

705

<210> 194

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-NF-6

<400> 194

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga catccagatg acacagagcc cgtcctccct gtctgcctct 120

gtgggagaca gagtcaccat cacctgcagg gcaagtcagg acattagtaa atatttaaat 180

tggtatcagc agaaaccaga cggaactgtt aaactcctga tctaccatac atcaagatta 240

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta caccctcacc 300

attagcagcc tgcaaccgga agatattgcc acttacttct gccaacaggg taatacgctt 360

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg agtcaggacc tggcctggtg 480

aaaccctcac agactctgtc cctgacatgc actgtctcag gggtctcatt acccgactat 540

ggtgtaagct ggattcgcca gcctccaggt aagggtctgg agtggctggg agtaatatgg 600

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat ctccaaggac 660

aactccaaga gccaagtttc cttaaaatta agtagtgtta ctgctgctga cacagccgtc 720

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 780

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 195

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-6

<400> 195

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 196

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-NF-7

<400> 196

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga catccagatg acacagagcc cgtcctccct gtctgcctct 120

gtgggagaca gagtcaccat cacctgcagg gcaagtcagg acattagtaa atatttaaat 180

tggtatcagc agaaaccagg tggaactgtt aaactcctga tctaccatac atcaagatta 240

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagattt caccctcacc 300

attagcagcc tgcaaccgga agatattgcc acttactact gccaacaggg taatacgctt 360

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg agtcaggacc tggcctggtg 480

aaaccctcac agactctgtc cgtgacatgc actgtctcag gggtctcatt acccgactat 540

ggtgtaagct ggattcgcca gcctccaggt aagggtctgg agtggctggg agtaatatgg 600

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat ctccaaggac 660

aactccaaga gccaagtttc cttaaaatta agtagtgtta ctgctgctga cacagccgtc 720

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 780

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 197

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-7

<400> 197

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 198

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-NF-8

<400> 198

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga catccagatg acacagagcc cgtcctccct gtctgcctct 120

gtgggagaca gagtcaccat cacctgcagg gcaagtcagg acattagtaa atatttaaat 180

tggtatcagc agaaaccagg tggaactgtt aaactcctga tctaccatac atcaagatta 240

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagattt caccctcacc 300

attagcagcc tgcaaccgga agatattgcc acttacttct gccaacaggg taatacgctt 360

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg agtcaggacc tggcctggtg 480

aaaccctcac agactctgtc cgtgacatgc actgtctcag gggtctcatt acccgactat 540

ggtgtaagct ggattcgcca gcctccaggt aagggtctgg agtggctggg agtaatatgg 600

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat ctccaaggac 660

aactccaaga gccaagtttc cttaaaatta agtagtgtta ctgctgctga cacagccgtc 720

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 780

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 199

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-8

<400> 199

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 200

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-NF-9

<400> 200

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga catccagatg acacagagcc cgtcctccct gtctgcctct 120

gtgggagaca gagtcaccat cacctgcagg gcaagtcagg acattagtaa atatttaaat 180

tggtatcagc agaaaccaga cggaactgtt aaactcctga tctaccatac atcaagatta 240

cactcaggag tcccatcaag gttcagtggc agtgggtctg gaacagatta caccctcacc 300

attagcagcc tgcaaccgga agatattgcc acttacttct gccaacaggg taatacgctt 360

ccgtacacgt tcggaggggg gaccaagctg gagatcacag gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg agtcaggacc tggcctggtg 480

aaaccctcac agactctgtc cgtgacatgc actgtctcag gggtctcatt acccgactat 540

ggtgtaagct ggattcgcca gcctccaggt aagggtctgg agtggctggg agtaatatgg 600

ggtagtgaaa ccacatacta taattcagct ctcaaatcca gactgaccat ctccaaggac 660

aactccaaga gccaagtttc cttaaaatta agtagtgtta ctgctgctga cacagccgtc 720

tactactgtg ccaaacatta ttactacggt ggtagctatg ctatggacta ctggggccaa 780

ggaacctcag tcaccgtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 201

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-9

<400> 201

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 202

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-NF-10

<400> 202

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccggg cggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgattt taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattatt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cgtgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgcgc aaaggcctgg aatggctggg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa gccaggtgag cctgaaaatg agcagcgtga ccgcggcgga taccgcgatt 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 203

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-10

<400> 203

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 204

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> DNA NK19H-NF-11

<400> 204

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccggg cggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgattt taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattttt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cgtgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgcgc aaaggcctgg aatggctggg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa gccaggtgag cctgaaaatg agcagcgtga ccgcggcgga taccgcgatt 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 205

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-11

<400> 205

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<210> 206

<211> 2150

<212> DNA

<213> Homo sapiens

<220>

<221> another sign

<223> NK19H-NF-12 DNA

<400> 206

gaattcgccg ccaccatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg 60

ctccacgccg ccaggccgga tattcagatg acccagagcc cgagcagcct gagcgcgagc 120

gtgggcgatc gcgtgaccat tacctgccgc gcgagccagg atattagcaa atatctgaac 180

tggtatcagc agaaaccgga tggcaccgtg aaactgctga tttatcatac cagccgcctg 240

catagcggcg tgccgagccg ctttagcggc agcggcagcg gcaccgatta taccctgacc 300

attagcagcc tgcagccgga agatattgcg acctattttt gccagcaggg caacaccctg 360

ccgtatacct ttggcggcgg caccaaactg gaaattaccg gtggcggtgg ctcgggcggt 420

ggtgggtcgg gtggcggcgg atctcaggtg cagctgcagg aaagcggccc gggcctggtg 480

aaaccgagcc agaccctgag cgtgacctgc accgtgagcg gcgtgagcct gccggattat 540

ggcgtgagct ggattcgcca gccgccgcgc aaaggcctgg aatggctggg cgtgatttgg 600

ggcagcgaaa ccacctatta taacagcgcg ctgaaaagcc gcctgaccat tagcaaagat 660

aacagcaaaa gccaggtgag cctgaaaatg agcagcgtga ccgcggcgga taccgcgatt 720

tattattgcg cgaaacatta ttattatggc ggcagctatg cgatggatta ttggggccag 780

ggcaccagcg tgaccgtgag cagcaccacg acgccagcgc cgcgaccacc aacaccggcg 840

cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 900

ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 960

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg ccggagggac 1020

cagaggctgc cccccgatgc ccacaagccc cctgggggag gcagtttccg gacccccatc 1080

caagaggagc aggccgacgc ccactccacc ctggccaaga tcagagtgaa gttcagcagg 1140

agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200

ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260

ggaaagccga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320

atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380

gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440

caggccctgc cccctcgcgg ctctggcgag ggaaggggtt ccctgcttac ttgcggcgac 1500

gtcgaagaga atcccggtcc gatggccctc ccagtaactg ccctcctttt gcccctcgca 1560

ctccttcttc atgccgctcg ccccaactgg gtcaacgtga ttagcgattt gaagaaaatc 1620

gaggacctta tacagtctat gcatattgac gctacactgt atactgagag tgatgtacac 1680

ccgtcctgta aggtaacggc catgaaatgc tttcttctgg agctccaggt catcagcttg 1740

gagtctgggg acgcaagcat ccacgatacg gttgaaaacc tcatcatcct tgcgaacaac 1800

tctctctcat ctaatggaaa cgttacagag agtgggtgta aggagtgcga agagttggaa 1860

gaaaaaaaca tcaaagaatt tcttcaatcc ttcgttcaca tagtgcaaat gttcattaac 1920

acgtccacta ccacacccgc cccgaggcca cctacgccgg caccgactat cgccagtcaa 1980

cccctctctc tgcgccccga ggcttgccgg cctgcggctg gtggggcggt ccacacccgg 2040

ggcctggatt ttgcgtgcga tatatacatc tgggcacctc ttgccggcac ctgcggagtg 2100

ctgcttctct cactcgttat tacgctgtac tgctaagcgg ccgcgtcgac 2150

<210> 207

<211> 706

<212> Protein

<213> Homo sapiens

<220>

<221> another sign

<223> amino acid sequence of NK19H-NF-12

<400> 207

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

20 25 30

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln

35 40 45

Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr

50 55 60

Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro

65 70 75 80

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile

85 90 95

Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly

100 105 110

Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

115 120 125

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

130 135 140

Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

145 150 155 160

Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly

165 170 175

Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu Gly

180 185 190

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

195 200 205

Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Ser Leu Lys

210 215 220

Met Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Ala Lys

225 230 235 240

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly

245 250 255

Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro

260 265 270

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu

275 280 285

Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp

290 295 300

Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly

305 310 315 320

Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Arg Asp Gln

325 330 335

Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly Gly Ser Phe Arg

340 345 350

Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser Thr Leu Ala Lys

355 360 365

Ile Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val

485 490 495

Glu Glu Asn Pro Gly Pro Met Ala Leu Pro Val Thr Ala Leu Leu Leu

500 505 510

Pro Leu Ala Leu Leu Leu His Ala Ala Arg Pro Asn Trp Val Asn Val

515 520 525

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

530 535 540

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

545 550 555 560

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

565 570 575

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

580 585 590

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

595 600 605

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

610 615 620

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Thr Thr Thr

625 630 635 640

Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro

645 650 655

Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val

660 665 670

His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro

675 680 685

Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu

690 695 700

Tyr Cys

705

<---

Claims (86)

1. A polypeptide that binds CD19, comprising a CD19-directed chimeric antigen receptor (CAR), wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence set forth in SEQ ID NO: 120; and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain. 2. The polypeptide according to claim 1, wherein the OX40 subdomain comprises the amino acid sequence under SEQ ID NO: 6. 3. The polypeptide according to claim 1 or 2, wherein the CD3-zeta subdomain comprises the amino acid sequence of SEQ ID NO: 8. 4. The polypeptide of any one of claims 1-3, wherein the hinge comprises a CD8-alpha hinge. 5. The polypeptide of any one of claims 1-4, wherein the CD8-alpha transmembrane domain comprises the amino acid sequence of SEQ ID NO: 4. 6. A natural killer (NK) cell for expressing a chimeric antigen receptor (CAR) comprising a polynucleotide encoding a polypeptide according to any one of claims 1-5. 7. A natural killer (NK) cell expressing a chimeric antigen receptor (CAR) containing: (a) a polynucleotide encoding a CD19-directed chimeric antigen receptor (CAR), wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence set forth in SEQ ID NO: 120; and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and an intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain; and (b) a polynucleotide encoding membrane-associated interleukin-15 (mbIL15) comprising the amino acid sequence set forth in SEQ ID NO: 12. 8. A natural killer (NK) cell expressing a chimeric antigen receptor (CAR) containing a polynucleotide encoding: (a) CD19-directed chimeric antigen receptor (CAR), wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence shown as SEQ ID NO: 120, and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and an intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain; and (b) membrane-associated interleukin-15 (mbIL15) comprising the amino acid sequence set forth in SEQ ID NO: 12. 9. The NK cell of claim 7 or 8, wherein the hinge comprises a CD8-alpha hinge. 10. The NK cell of any one of claims 7-9, wherein the CD8-alpha transmembrane domain comprises the amino acid sequence of SEQ ID NO: 4. 11. The NK cell of any one of claims 7-10, wherein the OX40 subdomain comprises the amino acid sequence of SEQ ID NO: 6. 12. The NK cell of any one of claims 7-11, wherein the CD3-zeta subdomain comprises the amino acid sequence of SEQ ID NO: 8. 13. Use of a polypeptide according to any one of claims 1-5 or an NK cell according to any one of claims 6-12 in the preparation of a medicinal product for the treatment of cancer in which CD19 is expressed. 14. Use of a polypeptide according to any one of claims 1-5 or an NK cell according to any one of claims 6-12 for the treatment of cancer in which CD19 is expressed. 15. The use according to claim 13 or 14, wherein the cancer is lymphoma. 16. A polynucleotide encoding a CD19-directed chimeric antigen receptor (CAR) for use in the treatment of cancer, wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence shown as SEQ ID NO: 120, and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain; wherein the polynucleotide further encodes membrane-associated interleukin-15 (mbIL15) comprising the amino acid sequence of SEQ ID NO: 12. 17. The polynucleotide of claim 16, wherein the OX40 subdomain comprises the amino acid sequence of SEQ ID NO: 6. 18. The polynucleotide according to claim 16 or 17, wherein the CD3-zeta subdomain comprises the amino acid sequence of SEQ ID NO: 8. 19. A method for producing a population of natural killer (NK) cells expressing a CD19-directed chimeric antigen receptor (CAR) for the treatment of cancer, the method comprising: (a) delivering to a plurality of NK cells a vector comprising a polynucleotide encoding a CD19-targeted CAR, wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence shown as SEQ ID NO: 120, a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; transmembrane domain of CD8-alpha and intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain; wherein the polynucleotide further encodes a membrane-bound interleukin-15 (mbIL15) comprising the amino acid sequence of SEQ ID NO: 12, thereby generating a population of engineered NK cells; and (b) expanding the engineered NK cells in culture to a population size suitable for the desired dose, thereby obtaining a population of NK cells expressing a CD19-directed chimeric antigen receptor for the treatment of cancer. 20. Use of a polypeptide comprising a CD19-directed chimeric antigen receptor (CAR) in the preparation of a medicinal product for the treatment of a cancer in which CD19 is expressed, wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence set forth in SEQ ID NO: 120; and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain. 21. Use of a natural killer (NK) cell comprising a polynucleotide encoding a CD19-directed chimeric antigen receptor (CAR) in the preparation of a medicament for the treatment of a subject having a cancer that expresses CD19, wherein the CAR comprises: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence set forth in SEQ ID NO: 120; and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain. 22. The use according to claim 20 or 21, wherein the cancer is leukemia. 23. The use according to claim 20 or 21, wherein the cancer is lymphoma. 24. The use according to any one of claims 20-23, wherein the NK cells are allogeneic to the subject. 25. A pharmaceutical composition for use in the treatment of a tumor in which CD19 is expressed, the composition comprising: a plurality of natural killer (NK) cells expressing a CD19-directed chimeric antigen receptor (CAR), wherein the CAR contains: extracellular CD19 binding fragment containing single-chain variable fragment (scFv) containing: a heavy chain variable domain (VH) comprising the amino acid sequence shown as SEQ ID NO: 120, and a light chain variable domain (VL) comprising the amino acid sequence shown as SEQ ID NO: 118; hinge; transmembrane domain of CD8-alpha and intracellular signaling domain containing the OX40 subdomain and the CD3-zeta subdomain. 26. The pharmaceutical composition according to claim 25, wherein the OX40 subdomain comprises the amino acid sequence under SEQ ID NO: 6. 27. The pharmaceutical composition according to claim 25 or 26, wherein the CD3-zeta subdomain comprises the amino acid sequence under SEQ ID NO: 8. 28. The pharmaceutical composition according to any one of claims 25-27, wherein the hinge comprises a CD8-alpha hinge. 29. The pharmaceutical composition according to any one of claims 25-28, wherein the transmembrane domain of CD8-alpha comprises the amino acid sequence under SEQ ID NO: 4. 30. The pharmaceutical composition according to any one of claims 25-29, wherein the plurality of NK cells additionally express membrane-associated interleukin-15 (mbIL15) comprising the amino acid sequence presented under SEQ ID NO: 12.