TW200920399A - Methods for screening antibodies and antibodies obtained therefrom - Google Patents

Abstract

It is intended to provide a novel screening method for an antibody, an antibody obtained by the method and a pharmaceutical composition containing the antibody as an active ingredient. The screening method for an antibody against a B-cell specific surface antigen, comprising the steps of: i) adding an antibody against a B-cell specific surface antigen to B cells; ii) confirming that IL-6 production by B cells is changed and/or confirming that IL-10 production by B cells is changed. The antibody which is against a B-cell specific surface antigen and has an action of changing IL-6 production by B cells and/or has an action of changing IL-10 production by B cells. The pharmaceutical composition containing the antibody as an active ingredient.

Description

200920399 IX. INSTRUCTION DESCRIPTION OF THE INVENTION [Technical Field] The present invention relates to an antibody screening method, which is an antibody against a surface antigen specific to B cells and has an adjustable interleukin by B cells (hereinafter Also referred to as "IL")-6, and/or have an effect of regulating the production of IL-10 by B cells, and an antibody obtained by the screening method and a medicine containing the antibody as an active ingredient. Composition [Prior Art] In recent years, antibody medicines using B cells as a target have been known to be effective for cancer treatment of non-Hodgkin's lymphomas and associated B cell lymphoid tumors. Among them, an anti-CD20 antibody or an anti-C 2 2 antibody having a B cell-specific surface antigen CD20 or CD22 as a target is reported, and an effective method for depleting B cells by such antibodies is effective. For example, Rituxan (registered trademark) is a chimeric/monoclonal antibody specific for the CD20 antigen of the B cell surface molecule (Non-Patent Document 1), and B cell selectivity can be eliminated for a long period of time (Non-Patent Document 2). This antibody has long been used for the treatment of non-Hodgkin's lymphoma (Non-Patent Document 3). Recently, an autoimmune disease has been reported to be an effective clinical test result showing that the B cell which is used as a target for the treatment of such cancers is effective. For example, in the clinical trials of patients with refractory chronic rheumatoid arthritis, the administration of sputum and methotrexate (Methotrexate 200920399) showed a greater improvement than that of methotrexate alone, and The effect continues (Non-Patent Document 4). On the other hand, there has been reported a clinical trial report on the effectiveness of Epratuzmab against primary Sjogren's syndrome in the anti-CD22f preparation (Non-Patent Document 5). These are thought to be caused by the disappearance of the autoimmune response due to the disappearance of the cause b cells. However, although the possibility of providing an inhibitory signal to B cells by the mechanism of autolysis of the autoimmune response of Epto is also discussed, that is, the phosphorylation-derived activity of CD22 is explored (Non-Patent Document 6), but The mechanism of action is currently unclear. Further, a method for treating an autoimmune disease by a novel anti-CD22 monoclonal antibody has been reported (Patent Documents 1 and 2). Although the results of clinical trials have been shown above, these antibody drugs have long-term continuous disappearance of B cells in the peripheral circulation for a long period of time. As a result, it becomes "B cells that are members of the body's defense function" and "B cells that control autoimmunity caused by the production of interleukin-10 in the direction of healing" (Non-Patent Documents 7 and 8). Lack of the risk of infection and cancer, but also reduce the natural healing power. Moreover, since these antibody medicines are very expensive treatments, the cost of the medicines becomes an obstacle to use. For example, in a clinical trial for patients with chronic rheumatic arthritis, a dose of 1 g as high as 1 g is administered (Non-Patent Document 4). In addition, in the clinical trials for the primary Shergar's syndrome, the Apocytium is administered in an amount of 360 mg/m 2 four times a week (Non-Patent Document 5), which is also a high dose, and becomes a patient economy. The burden on it. -6- 200920399 If you consider the above, the ideal antibody medicine system does not completely eliminate the B cells in the peripheral circulation, and can use B cells as the target antibody medicine to control the effect of the B cell function by a relatively low amount. These are considered to be achieved, in particular, by reducing the IL-6 production capacity of the causative B cells and/or by inducing or maintaining an antibody drug capable of producing B cells of IL-10 which are important for natural healing. As a mechanism, it is expected that the inhibitory receptor (CD2, Fc r RUB (CD32), CD72 and paired immunoglobulin-like substances present on the B cell membrane is intervened in the pathogenic B cells in an abnormally activated state. The receptor (PIR)-B or the like provides an inhibitory signal (Non-Patent Document 9) to induce B cells to a normal state. As described above, although antibodies which bind to B cell-specific surface antigens and which do not selectively degrade B cells or reduce their functions have been reported so far, in the treatment of autoimmune diseases, IL-6 is used. The screening method for the B cell surface antigen antibody, which is used as an indicator of IL-1 production and the production of IL-6 and/or IL-10, and the effective antibody as an indicator, has not been reported at all. Moreover, the agonist antibody for treating autoimmune diseases by providing an inhibitory signal to the pathogenic B cells of the abnormally activated state is completely unrelated, and the screening method for using a certain stage of the inhibitory signal transmission path as an index Nor has it been reported. [Patent Document 1] Japanese Laid-Open Patent Publication No. 2005-526501 [Non-Patent Document 1] Blood 83, 435-445 (1994) [Non-Patent Document 2] Arthritis Rheum 54, 6 1 3 - 620 (20 0 6) [Non-Patent Document 3] Curr Opin. Oncol. 1 0, 54 8-5 5 1 (1 99 8) 200920399 [Non-Patent Document 4] N Engl. J Med 3 5 0, 2572-25 8 1 (2004) [Non-Patent Document 5] American Rheumatology Association 2005 (speech No. 681, Society Transcription: S277) [Non-Patent Document 6] Clin Cancer Res 9, 3982s-3990s, (2003) [ Non-Patent Document 7] J Exp Med 184, 2271-2278 (1996) [Non-Patent Document 8] Nat Immunol 3, 944-950 (2002) [Non-Patent Document 9] Current Opinion in Immunology 17, 290-297 (2005) SUMMARY OF THE INVENTION [Problem to be Solved by the Invention] An object of the present invention is to provide a simple and effective screening method for providing an antibody against B cells as a target for the treatment of an autoimmune disease, the like, the provision of the antibody, and the provision of the antibody. The obtained antibody is used as a pharmaceutical composition of an active ingredient. [Means for Solving the Problem] The present inventors have found that an antibody against B cells which is effective for use in an autoimmune disease as a target has been found to have a regulatory effect on IL-6 production and/or to IL-10. It produces a regulatory antibody that does not cause peripheral B cells to disappear, and can control B cell function at a lower dose and can be used for the prevention of autoimmune diseases or B-cell lymphoid tumors. That is, the gist of the present invention is as follows. [1] An antibody which is anti--8-200920399 for a surface antigen specific to a B cell and which has a function of changing the production of interleukin-6 by B cells and/or has a whitening effect on B cells. The prime-1 0 produces an anti-ja^ body that undergoes a change. [2] The antibody according to [1], wherein the B cell-specific surface antigen is at least one selected from the group consisting of CD22a, CD22p, CD 19, CD20, CD79a and CD79P. [3] The antibody according to [1] or [2] wherein the B cell-specific surface antigen is CD22a and/or CD22P. [4] The antibody according to [3], wherein the CD22a and/or CD22P is derived from a human or non-human primate. [5] The antibody according to any one of [1] to [4] wherein the antibody against the B cell-specific surface antigen is a monoclonal antibody. [6] The antibody according to any one of [1] to [5] wherein the antibody against the B cell-specific surface antigen is a mouse, a non-primate or a human monoclonal antibody or any two of the antibodies A chimeric antibody composed of the species. [7] The antibody according to [6], wherein the antibody against the B cell-specific surface antigen is an isolated human antibody of the complete human type. [8] The antibody according to any one of [1] to [7] which is an antibody which inhibits the production of interleukin-6 by B cells and/or an antibody which enhances the production of interleukin-10 by B cells. [9] The antibody according to any one of [1] to [8] wherein the antibody against the B cell-specific surface antigen does not have a function of reducing normal B cells. [10] The antibody according to [9], wherein the normal B cell is reduced by 〇% or more and 2% or less by the antibody against the B cell-specific 200920399 surface antigen 11 [11] such as [3] to [1] The antibody according to any one of the preceding claims, wherein the extracellular regions 1 and 2 of cD22a and/or CD22p are not recognized by antibodies against CD22a and/or CD22P. [12] The antibody according to [11], wherein an epitope of an antibody against c D 2 2 a and/or CD22P is present between the extracellular regions 3 to 7 of CD22a and/or CD22p, respectively. [13] The antibody according to [12], wherein an epitope of an antibody against c D 2 2 a and/or CD22P is present between the extracellular regions 3 to 5 of cD22a and/or CD22P, respectively. [14] The antibody according to [12], wherein an epitope of an antibody against CD22a and/or CD22β is present between CD22 (extracellular regions 5 and 7 of x and/or CD22P, respectively). [15] An antibody that binds to an epitope that is determinable by an antigen that recognizes an antibody against CD22a and/or CD220 that is selected from the group consisting of: 1Β11, 12F6, 13B1 1, 17H5, 20F4, and 23C6. The same method. [1 6] An screening method for an antibody, which is a screening method for an antibody resistant to a B cell-specific surface antigen, the method comprising: i) adding a surface specific to the B cell to the B cell The step of the antibody of the antigen; U) the step of changing the IL-6 production by the B cell and/or confirming the step of changing the IL-10 production by the B cell. [1 7] The screening method according to [16], wherein the B cell-specific-10-200920399 surface antigen is selected from the group consisting of CD22a, CD22p, CD19, CD20, CD79 (x and CD7 9p) [18] The method according to [16] or [17], wherein the b cell-specific surface antigen is selected from the group consisting of CD22a and/or CD22P. [19] [16] to [18] The screening method according to the item, wherein the B cell line is selected from the group consisting of B cells, derived from rodent peripheral blood cells, derived from rodent lymphoid cells, derived from rodent spleen cells, derived from human peripheral blood cells, and sources. A screening method according to any one of [16] to [19], wherein the B cell is a human lymphocyte, a human spleen cell, and a human spleen cell. The interleukin-6 production is induced and/or enhanced by activation of B cells. [2] The screening method according to [20], wherein the activation of B cells is by a group selected from the group consisting of At least one of which is activated: anti-IgM antibody, anti-IgM antibody beads, anti-IgG antibody, anti-IgG antibody beads, anti-Ig antibody Anti-Ig antibody beads, bacterial elements of Staphylococcus aureus Cowan I strain, lipopolysaccharide, Pokeweed mitogen, Epstein-Barr virus, CpG [22] The method of screening according to any one of [16] to [21] wherein the step ii) inhibits B cell production of interleukin-6 And / or enhance B cells to produce interleukin-10. [23] The screening method as described in [22], in which inhibition of B cell production -11 - 200920399 Interleukin-6 system only causes the disappearance of b-cells producing interleukin-6 and/or only for the generation of interleukin B cells of -6 inhibit the production of interleukin-6. [24] The screening method according to any one of [16] to [23] wherein the change in the production of the interleukin-6 is suppressed to 5% or more and 100% or less. [25] The screening method according to any one of [16] to [23] wherein the change in the production of the interleukin-6 is inhibited at 25 % or more and 100% or less. [2] The screening method according to any one of [1 6 ] to [2 3], which confirms that the change in the production of the interleukin-6 is suppressed to 40% or more and 10% or less. [2] The method of detecting the hoof according to any one of [1 6 ] to [2 3], wherein the change in the production of the interleukin-10 is enhanced to 150% or more. [28] A method for producing an antibody produced by the method of [16] to [27] or the antibody of [1] to [1 5] by a genetic recombination method. [29] An antibody obtained by the method according to any one of [16] to [27]. [3] The antibody according to [29], wherein the anti-system monoclonal antibody. [31] The antibody according to [29] or [30] wherein the anti-systematic human monoclonal antibody. [32] A pharmaceutical composition comprising the antibody according to any one of [1] to [15] or [29] to [31] as an active ingredient. [33] The pharmaceutical composition according to [32], which is for treating and/or preventing a disease selected from the group consisting of a disease associated with interleukin-6 and/or interleukin-10 and a tumor of B lymphoma. [3 4] The pharmaceutical composition as described in [3 2] is used for the prevention and/or treatment of autoimmune diseases or Castleman's diseases by -12-200920399. [3] The pharmaceutical composition according to [3 2], which is for preventing and/or treating chronic rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus. [Effect of the Invention] According to the present invention, it is possible to provide a screening method for a novel antibody which can be used for the prophylaxis of autoimmune diseases without controlling the B cell function of the peripheral circulation, and which can control the B cell function at a low dosage, and the method can be obtained by the method. An antibody and a pharmaceutical composition containing the antibody as an active ingredient. [Embodiment] Hereinafter, the present invention will be described in detail. As a state of the present invention, a screening method for an antibody against a surface antigen specific to B cells, and exemplified by having a regulatory effect on IL-6 production by B cells and/or IL-10 production by B cells. Screening method for antibodies with regulatory effects. (1) B cell-specific surface antigen The B cell-specific surface antigen used in the present invention is a protein which can be expressed on the cell surface of B precursor cells or mature B lymphocytes. In the present invention, a known B-cell-specific surface antigen can be used. Preferred examples are CD10, CD19, CD20, CD21, CD22a, CD22p, CD23, CD24, CD37, CD38, CD39, CD40, CD72, CD73, CD7 4-13-200920399, CD79a, CD79P 'CD80, CD81, CD82, CD84, CD86 or CD139. More preferably, it is CD22a, CD22P, CD19, CD20, CD79a or CD79P. It is best to be CD22a or CD22P. The origin of the B cell-specific surface antigen in the present invention is not particularly limited, and may be suitably selected from human, non-human, mouse, rat or rodent, but preferably derived from human or non-human. Primates, preferably surface antigens specific to human B cells. CD22 is known to have two isoforms. One of the isomeric forms of CD22P is a membrane-through glycoprotein with a molecular weight of 1300-140 kDa, and its extracellular region is composed of seven domains (J. Exp. Med. 1 73, 1 3 7- 1 46 ( 1 99 1 ); J. Immunol. 1 50, 50 1 3 -5 024 (1 993 ), Annu. Rev. Immunol. 15, 48 1 -504 (1 997), J. Biol. Chem. 268 , 7019-7023 (1993)). The extracellular region of CD22P is shown in Figure 1 (A). On the other hand, the extracellular domain of the isoform CD2 2α lacks regions 3 and 4 of CD220 and is composed of five regions (Nature 344, 74-77 (1 990), J. Exp. Med. 1 73, 1 3 7- 1 46 (1 99 1 ), J. Biol. Chem. 273, 2789-27815 (1 998), Glycoconj. J. 13, 913-926 (1996)). In this application, CD22P is also referred to as "full length CD22". Further, the region 1 of the extracellular region of the full-length CD22 (CD22p) is amino acid number 22 to 138, the region 2 is the amino acid number 139 to 240, the region 3 is the amino acid number 241 to 3 29, and the region 4 is the amine. The base acid number is 33〇~417, the region 5 is the amino acid number 41 8~5 03, the region 6 is the amino acid number-14-200920399 504~591, and the region 7 is the amino acid number 592~679 〇.£\ Household · 173, 137-146 (1991)). The CD22a and β aliases are also known as 8184 (:-2, 81 §16〇2; 61^, MGC130020, FLJ22814, Β-lymphocyte antigen 8 (Β 抗原 antigen 8), Lyb-8 or Lyb8. Here, B The method for obtaining a cell-specific antigen protein may be, for example, a commercially available product, a recombinant product using a gene, or the like. As for the antigen-expressing cells of the B-specific antigen, the original B cell-specific antigen may be used or may be derived from the B cell-specific cells. The antigen expresses the plastid and extracts the cell-specific antigen expression. Further, the B cell-specific antigen-expressing plastid can be constructed by inserting a B-cell-specific antigen cDNA into an appropriate epitope to construct the B-specific antigen cDNA system. A cell-specific antigen cDNA selected from a suitable cell-modulated cDNA or a base DNA, a cDNA gene library, a genomic DNA gene library, or a DNA obtained from a known sequence such as a literature or a database, etc. At this time, B is obtained. The cell-specific cDNA may be a full-length cDNA, a cDNA fragment lacking a base, or a chimeric cDNA which may be a mouse or a human. (2) The antibody is obtained from the surface antigen specific to the B cell used in the present invention. antibody Known to be used as the surface antigen-specific antibodies in B cells of a good example is an anti-CD22cx antibody, anti-CD22P antibody, anti-CD19, anti-CD20 antibody, an anti-CD79a antibody or anti-CD79P antibody,

Med. CAM, the ball, the example, the high-B, the current B-cell, the B-imaging antigen, or just as long. Preferably, the antibody is -15-200920399 is an anti-CD22a antibody or an anti-CD22P antibody. Further, in the present invention, the anti-CD22 anti-system means anti-CD22a antibody and anti-CD220 antibody, and CD22 means CD22a and CD220 unless otherwise specified. The antibody to the B cell-specific surface antigen can be prepared by a commercially available product or a known method. For example, there is a method of collecting only a positive plasma-modulated high-valence preparation for an autoimmune disease patient, and a method for obtaining an antiserum from an animal immunized with a B cell-specific surface antigen protein or a B cell-specific surface antigen expression cell (Current Protocols) In Immunolgy, Unit 2.4, (1991)), a method for modulating a monoclonal antibody by culturing an antibody-producing cell (hybrid tumor) obtained by a cell fusion method (Current Protocols In Immunolgy, Unit 2.6-2.7, (1991)) A method for modulating a recombinant antibody by a genetic engineering method by a method for immunomodulating an antibody by a vector (Neurobiol. Dis. 14, 3 65 -3 79 (2 0 0 3 )) (Μ ο 1 · Cells 20, 1 7- 29 (2005), J. Immunother. 29, 1 -9 (2006)) et al. The antibody of the present invention is not particularly limited to binding to a B cell-specific surface antigen, and a mouse antibody, a rat antibody, a rabbit antibody, a goat antibody, a goat antibody, a camel antibody, or the like can be suitably used, but in order to make a heterologous to human The antigenicity reduction is preferably a chimeric antibody, a humanized antibody, a human antibody or the like. It is best to be a fully human type antibody. The fully human type antibody can be produced by a known method, and can be produced, for example, according to the method described in International Publication No. 01/025492 or International Publication No. 02/043478. Specifically, the transgenic non-human lactation containing the two human immunoglobulin gene loci can be made by -16-200920399 as described in International Publication No. 02/043478 or International Publication No. 02/043 478. The animal 'immunizes with a B cell-specific surface antigen, and the lymphocytes derived from the non-human mammal of the transgenic gene are fused with immortalized cells to form a hybridoma cell, thereby confirming the antibody produced by the hybridoma cell. A step of binding to a B cell-specific surface antigen, whereby a hybridoma that secretes a fully human type antibody of a human sequence can be produced. The hybridoma thus produced is cultured by a known method, and the secreted antibody is purified by a known method, whereby a fully human type antibody can be obtained. Alternatively, as described in Example 12, a fully human type antibody can also be obtained by a method using HuMAb-Mouse (registered trademark) or KM-Mouse (registered trademark). Further, the antibody of the present invention may be either a full-length antibody or an antibody fragment as long as it has binding ability to an antigen specific for B cells. Specific examples of the antibody fragment include Fab, Fab', F(ab') 2, Fv, scFv and the like. Further, the antibody of the present invention may be a conjugate antibody which binds to various molecules such as a polynucleotide, a radioactive substance or a toxin according to a known method. Further, the antibody of the present invention is preferably an individual. (3) Antibody type The antibody of the present invention contains any of a plurality of antibodies and monoclonal antibodies. Preferably, it is a monoclonal antibody (for example, a mouse, a non-human primate, a human monoclonal antibody, etc.), and more preferably a chimeric antibody consisting of two of these types (for example, including mouse and human origin) A portion of the chimeric antibody) is preferably a human monoclonal antibody. In the present invention, the antibodies selected and manufactured by the screening method are also the same. -17- 200920399 (4) B cell type The B cell used in the screening method of the present invention may be a B cell or a B cell in the primary culture. As the strain B cells, a known cell strain having IL-6 production ability and/or il-1 0 production ability is preferred, and Ramos (ATCC NO. CRL-1596) and Ramos. 2G6 4C10 (ATCC) are preferably used. NO. CRL- 1 923 ), RPMI 1 788 (ATCC NO. CCL-156) 'Raji (ATCC NO. CCL-86). As a primary culture, B cells are derived from rodent peripheral blood cells, from rodent lymphoid cells, from rodent spleen cells, from human peripheral blood, from human lymph nodes, from human spleen cells, or from humans. B cells of the tonsils. Preferably, it is a B cell derived from the blood of human peripheral blood. Further, whether or not the IL-6 or IL-1 〇-producing cell line is examined by an enzyme-linked immunosorbent assay (ELISA) method as shown in the examples below. (5) B cell depletion (B cell depletion) In the present invention, the B cell depletion action or the B cell killing action means that when the antibody of the present invention is added to the B cell, the antibody is caused by killing the B cell. The number of B cells is reduced. Normal B cells are important in the defense of the body, and if they are not indiscriminately killed, there is a risk of inducing severe infection. In the present invention, an antibody which is not destructive to normal B cells is preferred. More preferably, the antibody which only causes the IL-6-producing B cells to disappear. The B cell depletion action or B cell killing action in the present invention can be confirmed by the change in the number of B cells coexisting with the antibody of the present invention. B cells -18- 200920399 The number of B cells was directly calculated by a microscope, and the method of analyzing the B cell marker gene using the flow cytometry shown in the examples was also carried out. As a method of adding the antibody of the present invention to B cells and investigating the B cell depletion action or the B cell killing action of the antibody, there is a method described in Example 7 or 14. Further, there is a method in which a B cell of the present invention is added to a cultured B cell and cultured, and the damaged B cell is quantified by a known method. In the case of culturing, the method of culturing in the case of culturing on the second day of the invention is also carried out in a short period of time such as 24 hours, 12 hours, 6 hours, 3 hours, 1 hour or the like. In the present invention, it is preferred that the antibody of the present invention reduces the number of sputum cells to from 〇% to 40%. More preferably, the number of cells is reduced from 〇% to 30%, and more preferably 0 °/. Up to 20%. It is best to reduce the number of cells to 〇% to 1%. (6) IL-6 In the present invention, IL-6 is an abnormal expression in autoimmune diseases such as chronic rheumatoid arthritis (RA) and multiple bone marrow tumors, and is one of the causative substances of such pathological conditions ( Eur. J. Immunol. 18,1 797- 1 80 1 (1988)). As for the method for measuring the amount of IL-6 produced, there is a method using ELISA as described in the Examples below. Others, by using the flow cytometry flow cell microsphere array kit (Cyt 〇 metric B ead A rray K it ) ( BD Pharmingen ), using mouse B cell hybridoma strain, B9 cells Biological Analysis Methods (Current Protocols In Immunology, -19- 200920399

Unit 6.6.1-6.6.4, (1991)), etc. Determine the amount of IL-6 produced. (7) IL-10 In the present invention, IL-1 0 is the same factor for inhibiting the production of interleukin from Th 1 cells (J Exp. Med. 170, 2081-2095 (1989)) and is known to be An autoimmune disease such as multiple sclerosis (MS) or RA has a prophylactic therapeutic effect (Eur. J Immunol 23, 1745-1751 (1 993), J Immunol. 164, 1 576- 1 5 8 1 (2000) )). The method for measuring IL-10 was carried out by the same method as the above-described IL-6 production by a method using ELISA or a flow cytometer flow cytometry microarray kit (BD Pharmingen). (8) Changes in IL-6 production In the present invention, the change in IL-6 production includes any of inhibition of IL-6 production and induction and/or enhancement of IL-6 production. The production of IL-6 specifically means induction and enhancement by activation of B cells, which are anti-IgM antibodies, anti-IgM antibody beads, anti-IgG antibodies, anti-IgG antibody beads, anti-Ig antibodies, anti-Ig Antibody beads, Staphylococcus aureus Cowan I strain bacterial composition (SAC), lipopolysaccharide (LPS), Pokeweed fruit mitogen (

Pokeweed mitogen) (PWN), human vesicular virus (Epstein-

Barr virus ) ( EBV ) and other viruses and ion channels are activated! 1 (A23 1 8 7 ionophore, calcium ionophore, etc.), protein kinase C activator (Phorbol ester, etc.), for B cell surface molecules Anti--20- 200920399 (anti-CD20 antibody, anti-CD40 antibody, etc.), cells expressing B cell surface dominating ligand (CD40L, etc.), CPG oligodeoxynucleotides, Toll receptor ligands, Various lymphatic media (lymphokine) (IL-1, IL-2, IL-4, IL-5, IL-6, IL·10, IL-13, IN F-7, INF-α, INF- /3, TGF-etc.), 8-mercaptoguanine nucleoside (8-MG), dextran sulfate, poly(IC), indoleamine, etc. Commercial products can also be used for each of these. Preferably, activation of B cells means anti-IgM antibody, anti-IgM antibody beads, anti-IgG antibody, anti-IgG antibody beads, anti-Ig antibody, anti-Ig antibody beads, SAC, LPS, PWN, EBV, etc. Virus, ion channel activator (A23 1 87 ionophore, calcium ionophore, etc.), protein kinase C activator (phorbol ester, etc.), CpG oligodeoxynucleotide and/or Toll receptor ligand, etc. Made. Preferably, activation of B cells means anti-IgM antibody, anti-IgM antibody beads, anti-IgG antibody, anti-IgG antibody beads, anti-Ig antibody, anti-Ig antibody beads, SAC, LPS' PWN, EBV, etc. The virus, CpG oligodeoxynucleotide and/or Toll receptor ligand are prepared. As an anti-IgM antibody, an anti-human I gM polyclonal antibody of goat, rabbit, sheep, cow, horse, camel, cock, pig, monkey, woodchuck, hamster, mouse or rat is used, or mouse is used. Anti-human IgM monoclonal antibody to rat, hamster, rabbit, goat, sheep or monkey. As the antibody, mouse anti-human IgM F(ab') 2 is preferably used. The CpG oligodeoxynucleotide system recognizes the sequence of the DNA derived from bacteria by high frequency, and increases the stimulation from the B cell receptor by interrogating the Toll-like receptor ligand 9 (J. Exp. Med. 195, -21 - 200920399 1507-1512 (20 02)) to induce or enhance IL-6 producers. The method for activating B cells is selected according to the above-mentioned stimulant concentration or cell number, and can be carried out, for example, by adding a solution containing a stimulating agent to a B cell culture solution. In the present invention, it is preferred to inhibit IL-6 production by B cells by an antibody against a surface antigen specific to B cells, and it is particularly preferable to inhibit IL-6 production only by B cells producing IL-6, or by only B cells producing IL-6 are eliminated and IL-6 production is inhibited. Inhibition of IL-6 production is achieved by inhibiting the amount of IL-6 secreted from B cells. These include inhibition of transcriptional activity of the IL-6 gene, inhibition of the amount of IL-6 protein synthesis, and inhibition of secretion from cells of the IL-6 protein. The disappearance of only B cells producing IL-6 is achieved by causing B cells producing IL-6 to disappear due to necrosis or apoptosis. Necrosis is caused by exogenous (antibody-dependent cell-mediated cytotoxicity (ADCC), complement, toxin, anaerobic state, etc.), and the cells are initially swollen and the membrane is changed by autolysis. Apoptosis is a form of cell death induced by the autologous destruction process of the originally written gene, and is characterized by nuclear DNA decomposition, nuclear degeneration or condensation, and gluttony of cell debris. Necrosis or apoptosis can be achieved by using Ann exin V, biotin cell apoptosis detection kit (R&D Systems, Inc.), etc., in the cell membrane of phosphonyl-serine seen during apoptosis. Distribution changes by fluorescence or biotin-labeled Annexin V to induce apoptosis and stain the cells, so that the necrotic cells are stained with red fluorescent DNA fluorescent staining agent iodide-22-200920399 (Propidium Iodide) ) staining, each of these cells was analyzed by flow cytometry and detected. In the present invention, the amount of IL-6 produced prior to the addition of an antibody against a B cell-specific surface antigen to B cells is preferably 5% or more and 100% or less after the addition of the antibody. This means that when the amount of IL-6 produced by the B cells before the addition of the antibody is 1.0%, the amount produced is reduced to 95% to 〇% after the addition of the antibody. Preferably, the amount of IL-6 produced before the addition of the antibody to the b cell is inhibited by 25% or more and 10% or less after the addition of the antibody. This means that when the amount of IL-6 produced by the B cells before the addition of the antibody is 100%, the amount produced is reduced to 75% to 0% after the addition of the antibody. More preferably, the amount of IL-6 produced before the addition of the antibody to the b cell is inhibited by 40% or more and 100% or less after the addition of the antibody. This means that when the amount of IL-6 produced by the B cells before the addition of the antibody is 1.0%, the amount produced is reduced to 60% to 0% after the addition of the antibody. (9) Changes in IL-10 production The changes in the so-called IL-10 production in the present invention include any of inhibition of IL-10 production and induction and/or enhancement of IL-10 production. In the present invention, it is preferred that the antibody to the surface antigen specific for B cells is induced and/or enhanced by IL-10 production by B cells. Induction and/or enhancement of IL-10 production is achieved by inducing and/or enhancing the amount of IL-10 secreted from B cells. These have the induction and/or enhancement of transcriptional activity of IL-1 0 gene, induction and/or enhancement of IL-10 protein synthesis amount, -23-200920399, and secretion enhancement of IL-10 protein from cells. In the present invention, it is preferred that the amount of IL-10 produced before the addition of the antibody against the surface antigen specific to the sputum cell is increased by 1.5 times or more after the addition of the antibody. The greater the increase in the amount of IL-10 production, the better, for example, is preferably 2, 10, 50 or 1 fold of the amount of IL-10 produced by the sputum cells prior to the addition of the antibody. Further, the step of "confirming the change of IL-6 production by sputum cells and/or confirming the change of IL-1 0 production by sputum cells" means confirming the step of changing IL-6 production by sputum cells. The step of confirming the change of IL-10 production by the 、 cell or the step of confirming the change of IL-6 production by the sputum cell also includes the step of confirming that the IL-10 production of the sputum cell changes. Preferably, it comprises a step of confirming inhibition of IL-6 production by sputum cells, confirming a step of inducing and/or enhancing the production of IL-10 by sputum cells, or confirming inhibition of IL-6 production by sputum cells. The step also includes the step of confirming the induction and/or enhancement of IL-10 production by the sputum cells. Preferably, it is meant to include a step of confirming that IL-6 production by sputum cells is inhibited and that induction and/or enhancement of IL-1 0 production by sputum cells is confirmed. (1) Confirmation method for confirming changes in IL-6 production or IL-10 production, for example, a method for confirming changes in transcriptional activity of individual genes, a method for confirming changes in the amount of synthesis of each protein, and confirming the amount of secretion of each protein from cells. The method of change, etc. -24- 200920399 As a method for confirming a change in transcriptional activity, for example, a method confirmed by reporting a gene analysis according to a general method is exemplified. Specifically, a method for producing a vector by introducing an appropriate reporter gene such as luciferase or the like downstream of the promoter region of the IL-6 gene or the IL-10 gene, and introducing the vector into B cells for measuring luciferase activity . As a method for confirming the change in the amount of protein synthesis, in order to homogenize B cells, a method of measuring the amount of IL-6 or IL-10 contained therein by enzyme-linked immunosorbent assay (ELIS A) is used. As a method for confirming the change in the amount of protein secreted from the cells, a method of measuring the amount of IL-6 or IL-10 contained therein by using E LI S A is to take a culture solution of B cells. A specific method by ELISA is exemplified by the method shown in Example 10. (11) Epitope The so-called epitope is a specific structural unit that recognizes the binding antigen of an antibody. In the present invention, the epitope of the antibody of the present invention is not present in the region 1 or 2 of the extracellular region of CD22a and/or CD22/3, that is, it is preferred that the antibody does not recognize the region 1 or 2. More preferably, the epitope of the antibody of the present invention is present in the region of the extracellular regions 3 to 7. Preferably, the epitope of the antibody of the present invention is present in the region of the extracellular region 3 or 4, and is preferably present in the region of the extracellular region 5 to 7. When the screening method according to the present invention is obtained as an anti-human CD 2 2 α and/or CD22 0 antibody and inhibits IL-6 production by sputum cells and/or enhances il_10 production, the antibody obtained is The antibody to which the binding epitope is -2520 200920399 is also suitable as the antibody of the present invention. In the present invention, an antibody capable of recognizing an epitope which is an antigen recognizing an anti-human CD2 2 fully human type monoclonal antibody produced by hybridoma 1B11, 12F6, 13B11, 17H5, 20F4 or 23C6 is preferred. The decision base is the same. In addition, anti-human CD22 fully human type monoclonal antibodies (also referred to as 1B 11 , 12F6, 13B11, 17H5, 20F4 or 23C6 antibodies) produced by hybridoma 1B11, 12F6, 13B11, 17H5, 20F4 or 23C6 are also Preferred antibodies in the invention. (1 2) Screening method The method of screening the antibody of the present invention is carried out, for example, by the following. That is, various human B cell-specific antibodies were added to human peripheral blood single cells (PBMC), and an appropriate amount of anti-human IgM F(ab')2 was added, and cultured in a CO 2 incubator at 37 ° C for 48 hours. The amount of IL-6 produced and the amount of IL-10 produced in the supernatant were measured. As a result of measuring the IL-6 concentration and/or the IL-10 concentration as described above, it was judged that the anti-system of the present invention regulates IL-6 production and/or regulates IL-10 production. Screening of selected antibodies according to the present invention has therapeutic and/or prophylactic effects on diseases selected from diseases associated with IL-6 and/or IL-10 and B-cell lymphoid tumors, and is effective for prevention and/or treatment. Body immune disease or B cell lymphoma and the like. Further, examples of the autoimmune diseases include chronic rheumatoid arthritis (R A ), multiple sclerosis (M S ), and systemic lupus erythematosus (SLE). -26- 200920399 (1 3 ) Method for producing antibody In the present invention, once an antibody is selected by screening, an antibody can be produced according to a general method. That is, a method of producing an antibody by a culture of a hybridoma producing an antibody, producing a recombinant antibody or the like according to a genetic engineering method. Specifically, a method of producing a target antibody by inserting all or a part of cDN 具有 having a base sequence of an antibody selected by the screening method of the present invention into a recombinant vector, and introducing a recombinant vector. The recombinant vector used in the production of the antibody of the present invention is also preferably a vector which can be expressed in prokaryotic cells such as coliforms (for example, pBR3 22, pUC 1 19 or its like), but is preferably true. A vector which can be expressed in a nucleus cell, and a vector which can be expressed in a cell derived from a mammal, for example, a plastid vector described in the examples, pcDNA3.1 (Invitrogen), pDON-AI DNA (TaKaRa), etc. . The transformant used in the production of the antibody of the present invention may also be a coliform-like prokaryotic cell, preferably a eukaryotic cell, more preferably a mammalian-derived cell, as a mammal-derived cell, for example, China. Hamster ovary cells (CHO cells) and the like. (1 4) Specific antibody of the antibody The antibody of the present invention can be specified by an amino acid sequence or a polynucleotide sequence of a Complementarity-determining region (hereinafter referred to as CDR or CRD) of a light chain and a heavy chain. . The light chain and heavy chain CDRs derived from the antibody of the present invention can be recognized as a novel antibody other than the antibody which binds to the framework region of the human antibody from the source -27-200920399, which is also an antibody of the present invention. The antibody of the present invention preferably has at least 70 °/ of the light chain and heavy chain C D R s amino acid sequence of the antibody. Preferably, 80%, more preferably 95% 'more preferably 97% of the antibodies of the same sequence of CDRs. Alternatively, an antibody comprising a CDR sequence consisting of an amino acid sequence substituted, appended or deleted from one or more amino acids of the light chain and heavy chain CDRs amino acid sequences of the antibody is preferably inhibited An antibody produced by IL-6 from B cells and/or enhancing production of IL-1 〇. (1) Pharmaceutical composition The antibody of the present invention or the antibody according to the method of the present invention can be used as a prophylactic and/or therapeutic agent, and can be a pharmaceutical composition formulated and administered according to a known method. For example, a pharmaceutical composition directly as a liquid or as a suitable dosage form can be administered orally or parenterally to humans or mammals. The amount of the antibody obtained by the method according to the present invention to humans is, for example, 0.01 mg/kg to 50 mg/kg. Further, the dosage is based on age, body weight, general state of health, sex, diet, duration of administration, method of administration, rate of excretion, combination of drugs, degree of condition at the time of treatment of the patient, and consideration of these or other factors. It is decided that the antibody of the present invention or the antibody obtained according to the method of the present invention can be used for the prevention and/or treatment of diseases associated with IL-6 and/or IL-10, B lymphoma, autoimmune disease or Castlemani. )disease. Also, -28-200920399 is an autoimmune disease, and examples include chronic rheumatoid arthritis, multiple sclerosis, or systemic lupus erythematosus. [Examples] Hereinafter, the present invention will be specifically described by way of examples, but the present invention is not limited to the following examples. Example 1: Production of recombinant mouse C D 2 2 high-performance cells Mouse CD22-expressing cells were prepared by genetic engineering methods as follows. First, mRNA was extracted from mouse B lymphoma cell line WEHI23 1 cells (ATCC NO. CRL-1702) using a Quick Prep Micro mRNA purification kit (manufactured by Amersham Pharmacia Biotech). RT-PCR was performed using the RNA PCR kit (AMV) Ver. 2 · 1 (TaKaRa B i )) using the Random 9 mers attached to the kit. Subsequently, a mouse CD22 sequence-specific primer (SEQ ID NO: 1 and 2) and Pyrobest DNA polymerase (manufactured by TaKaRa Bio) were used for PCR reaction, and an amplified 2.8 kb fragment was isolated, followed by a DNA sequencer (Amersham Pharmacia). The company) determines the base sequence (SEQ ID NO: 3 and 4) by the Dideoxy method using the primer of the camping light marker. This fragment was designated as pNT48 as a mouse CD22 cDNA 'selected into pPO2.1-TOPO of a TOPO YA selection kit (manufactured by Invitrogen). Further, the mouse CD22 cDNA fragment was inserted into an animal cell expression vector to construct PCD22/mouse. Using Lipofectamine 2000 (manufactured by Invitrogen), pCD22/mouse was transfected into Chinese hamster ovary cells -29-200920399 strain CHO-K1 cells to obtain a transformant. The mouse CD22 high expression cells (Current Protocols In Immunology Unit 2.1) were selected by cell ELISA, and re-selected by super-dilution. The most stable high-performance strain was named rCD22-CHO#34. Example 2: Preparation of hamster anti-mouse CD22 antibody Primary immunization A recombinant mouse CD22 high-expression cell (rCD22-CHO#34) was mixed with an adjuvant (FCA) in a 1:1 mixture to prepare a cell of 1χ107 cells/head. The Armenian hamster was given subcutaneously. After the second time, the antigen was diluted with physiological saline and immunized several times. After the antibody valence was measured, the individual whose antibody valence was increased was boosted by intraperitoneal administration of rCD22-CH〇#34. Armenian hamster spleen cells were subjected to cell fusion with mouse myeloma cell line P3U1 in a polyethylene glycol (PEG) method (Kohler G, Milstein C Nature 256: 495-497, 1975). The cells cultured with the HAT selection after fusion were screened, and the mouse CD22 monoclonal antibody producing cell (hybrid tumor) (Current Protocols In Immunology Unit 2.5) was selected. Finally, seven kinds of anti-mouse CD22 monoclonal antibodies against the mouse CD22-specific reaction (18 strains of the secondary strain) were determined to produce hybridoma. Preparation of various anti-mouse CD22 monoclonal antibodies by general purification method from the culture supernatant obtained by culturing the hybridoma Example 3: Production of recombinant human CD22 high-performance cells The human CD22-expressing cells were prepared by genetic engineering methods as follows. -30- 200920399. First, human c22 sequence-specific primers (SEQ ID NOs: 5 and 6) and Pyrobest DNA polymerase (manufactured by TaKaRa Bio) obtained from a commercially available cDN A gene library derived from human lymphoid tumor cells (manufactured by Invitrogen) were used for P c . R reaction, after amplifying the amplified 2.6 kb fragment, using a DNA sequencer (Amersham Pharmacia or Applied Biosystem), using a fluorescently labeled primer for Dideoxy or double termination ( The Diterminator method determines the base sequence, but considers the known human CD22 sequence (SEQ ID NO: 7 and 8). This fragment was designated as human CD22 and cloned into pCR-Bluntll-TOPO of the Blunt TOPO PCR selection kit (Invitrogen) and designated as pNT62. Further, the human CD22 cDNA fragment was inserted into an animal cell expression vector pcDNA3.1Zeo( + ) (manufactured by Invitrogen) to construct pTM025. PTM02 5 was transfected into mouse myeloma cell line N S 0 cells using Lipofectamine 2000 (manufactured by Invitrogen) to contain 10 ° /. FBS and 250//g/mL Zeocin DMEM medium were used as the selection medium to obtain a transformant. The human CD22 high-performance cell line was selected in the same manner as in Example 1, and was selected by super-dilution. The most stable high-performance strain was named G11-14. The human CD22 expressing the selected strain was of a full human type (Fig. 1(A)). Next, regions 1, 2 (J Immunol. 1 49: 264 1 - 2649, 1 992 ) of mouse CD22 and regions 3 to 7 of human CD22 were prepared as follows (J Exp_ Med. 173: 137-146, 1991) The first mouse-human-hybrid CD22 expressing cells are combined. Using the primer shown in SEQ ID NO: 9 and the primer shown in SEQ ID NO: 10 with a phosphate group attached to the 5' side, a PCR reaction was carried out using the plastosome 48 of mouse CD22 of -31 - 200920399 as a template to obtain a small reaction. DNA fragments of regions 1 and 2 of murine CD22 were digested with Hindlll. Further, a PCR reaction was carried out using pTM02 5 as a template using primers shown in SEQ ID NO: 1 1 and SEQ ID NO: 1, and a DNA fragment containing regions 3 to 7 of human CD22 was obtained and digested with PstI. The two fragments were combined with a vector fragment obtained by digesting pcDNA3.1Zeo(+) (manufactured by Invitrogen) with Hindlll and PstI to construct a first mouse which binds regions 1 and 2 of mouse CD22 to regions 3 to 7 of human CD22. Human-hybrid CD22 expresses the plastid pTM029. Subsequently, pTM029 was transfected into mouse myeloma cell line NS0 cells using L i p o f e c t am i n e e 2 0 0 (manufactured by Invitrogen) to obtain a transformant. In the same manner as in Example 1, the first mouse-human-hybrid CD22 high-performance cells were selected and selected by super-dilution to obtain two strains of the highest stable strains, which were named 34-1/30 and B4-, respectively. 152. The CD22 showing the selected strain was a hybrid type of regions 1 and 2 of the mouse and regions 3 to 7 of the human (Fig. 1 (B)). Further, a second mouse-human-hybrid CD22 expressing cell in which the regions 1 and 2 of mouse CD22 and the regions 5 to 7 of human CD22 were bound was prepared as follows. The primer shown in SEQ ID NO: 9 and the primer shown in SEQ ID NO: 10 with a phosphate group attached to the 5' side were subjected to a PC R reaction. A DNA fragment containing regions 1 and 2 of mouse CD 22 was obtained and digested with Hindlll. . Further, a PCR reaction was carried out using primers shown in SEQ ID NO: 13 and SEQ ID NO: 14, and a DNA fragment containing regions 5 to 7 of human CD22 was obtained and digested with PstI. These fragments were combined with the vector fragment -32-200920399 obtained by digesting pcDNA3.1Zeo(+) (manufactured by Invitrogen) with Hindlll and PstI to construct a region 2 of mouse CD 2 2 and a region of human CD 2 2 The second mouse-human-hybrid CD12 that binds to 7 expresses the plastid pTM027. Subsequently, PTM027 was transfected into mouse myeloma cell line NS0 cells using Lipofectamine 2000 (manufactured by Invitrogen) to obtain a transformant. The second mouse of interest was selected in the same manner as in Example 1. The human-hybrid CD22 high-expression cell was selected by super-dilution to obtain the most stable high-performance strain, and was named B9_4. The CD22 showing the selected strain was a hybrid type of the region 1 and 2 of the mouse and the region 5 to 7 of the human (Fig. 1 (C)). Example 4: Preparation of a mouse anti-human CD22 antibody Recombinant human CD 2 2 High-performance cells G 1 1 -1 4 were suspended in physiological saline as 1 X 1 〇7 cells/1 〇〇VL, and BALB/c mice (5 weeks old, female) with F cr R11 b deficiency (Nature 3 79 :346-349 1 996 ) Subcutaneous immunization 4 times. The recombinant human CD22 high-performance cells 34-1 / 30 and B9-4 were treated with mitomycin C (1 50 // g/mL, 60 minutes at 30 ° C), and then suspended in physiological saline to 1 X. 1 〇7 cells/1 00 β L 'Immune to the Fc 7 RIIb-deficient BALB/c mice (5 weeks 齢, female) were immunized 11 or 15 times, respectively. After the antibody titer was determined in the mice, the human CD 22 high-performance cells (G Π -1 4, 3 4 - 1 / 3 0, B 9 - 4 ) were administered intraperitoneally to the individuals whose antibody prices were elevated. Add it. Mouse spleen cells were taken and subjected to cell fusion by PEG method (Nature 25 6:495 -497, 1 975) with mouse myeloma cell line X63 · Ag8 cells or P3U1 cells. In the same manner as in Example 2, cell culture supernatant-33-200920399 subjected to HAT selection after fusion was used for screening, and anti-human CD22 or anti-mouse/human-hybrid CD22 monoclonal antibody-producing cells (hybrid tumor) were selected. Next, 126 mouse anti-human CD22 monoclonal antibodies that specifically react with human CD22 were determined to produce hybridomas. Each mouse anti-human C D 2 2 monoclonal antibody was prepared by culturing the culture supernatant obtained by culturing the hybrid tumor according to the general purification method (Current Protocols In Immunology Unit 2.6-2.7). These antibodies are then referred to as "mouse anti-human CD22 monoclonal antibodies". Example 5: Screening of anti-mouse CD22 antibody using local lymph node cells of mouse experimental autoimmune encephalomyelitis (EEE) model with IL-6 production inhibitory activity and IL-10 production-inducing activity as an indicator Detection system (1) Construction of mouse experimental autoimmune encephalomyelitis (EAE) model due to the use of mouse experimental autoimmune encephalomyelitis (EAE) model as multiple sclerosis (MS) The model 'is therefore produced according to the report for the experiment (Nature 3 1 7:3 5 5 -3 5 8,1 98 5 ). That is, a suspension of MBP Acl-oxime composed of amino acids 1 to U of mouse bone marrow basic protein (MBP) and complete adjuvant H37RA (Difco), PL/JX SJL/J F1 mice (The

Jackson Laboratory, 10 weeks old, female) was immunized subcutaneously once, and pertussis toxin (200 ng) was given intravenously on day 2 and day 2 (List

Pharmaceuticals) induces EAE. (2) Method of administration The EAE model animal was administered with anti-mouse CD22 antibody A by tail vein once a day for 5 days at a dose of 0.2 to 200/xg/head, self-immunization-34-200920399. , B or C ( N = 4 ). The above antibody was diluted with physiological saline and used. (3) Cell preparation method Measurement of IL-6: On the 10th day from the start of immunization, local lymph nodes were taken for each group of animals to prepare a cell suspension (N = 4). Add antigen (MBP Acl-1 1,30 // g/mL) to lymph node cells (5xl〇6 cells/mL) of each group, and start by ELISA (Mouse IL-6 BD OptEIA ELISA set, BD) The IL-6 was cultured in the supernatant after 48 hours of stimulation. Measurement of IL-1: On the 10th day after the start of immunization, local lymph nodes were taken for each group of animals to prepare a cell suspension (N = 4). Anti-Mu (10 v g/mL) + anti-CD40 (1 μg/mL) was added to each group of lymph node cells (5×10 6 cells/mL) to selectively stimulate B cells. IL-10 in the culture supernatant after 48 hours of stimulation was measured by ELISA (Mouse IL-10 BD OptEIA ELISA set, BD). (4) Results IL-6 production from local lymph node cells by antigen stimulation was strongly inhibited by the anti-mouse CD22 antibody A, B or C (Fig. 2). On the other hand, IL-1 produced from local lymph node cells was produced in a dose-dependent manner several times by anti-mouse CD22 antibody B (Fig. 3). From the above, it was determined that an anti-mouse CD22 antibody exhibiting IL-6 production inhibitory activity and an anti-mouse CD22 antibody exhibiting IL-10 production-inducing activity can be identified by using the screening system. -35- 200920399 Example 6: Effect against mouse mouse CD22 antibody against mouse EAE model Anti-mouse CD22 antibody A having IL-6 production inhibitory ability as shown in Example 5 and simultaneous inhibition of IL-6 production The anti-mouse CD22 antibody with the ability and IL-10 inducing ability was administered to the EAE model mice, and the therapeutic effect was explored by using the survival rate as an index. (1) Construction of mouse EAE model In the same manner as in Example 5, EAE-infected mice were constructed. (2) Method of administration The anti-mouse CD22 antibody shown in Example 5 was administered to the EAE model animal at a dose of 2 to 200 #g/head, once a day from the immunization, and within 5 days by the tail vein. A or B (N=10). The above antibody was diluted with physiological saline and used. (3) Results The results are shown in Fig. 4. The two antibody mouse CD22 antibodies A and B, which exert the same inhibitory effect on IL-6 production, exert the same effect on EAE. By administering an anti-mouse CD22 antibody, the effect of EAE persistence relative to the control group. The anti-mouse CD22 antibody (Fig. 4 (B)), which has both IL-6 production inhibitory ability and IL-10 inducing ability, has an anti-mouse CD22 antibody A with only IL-6 production inhibition (Fig. 4 (A) )), showing that the lower amount can play a meaningful effect on EAE. Example 7: Effect of anti-mouse CD22 antibody on lymphocyte subtypes in mouse EAE model -36- 200920399 To investigate whether anti-mouse CD22 exerts effect on B cell disappearance, explore anti-CD22 The effect of antibodies on the lymphocyte subgroup. (1) Construction of mouse EAE model In the same manner as in Example 5, EAE-infected mice were constructed. (2) Method of administration The EAE model animal was administered with an amount of 2 000 vg/head, once a day from the immunization day, and administered by tail vein within 5 days, and inhibited by IL-6 production as shown in Example 5. Anti-mouse CD22 antibody C (N = 4). The above antibody was diluted with physiological saline and used. (3) Flow cytometry analysis of local lymph node cell suspensions of each group on the 1st day of immunization, using rat phycoerythrin (PE)-labeled rat anti-mouse B220 monoclonal antibody (BD) Pharmingen) As a B cell marker gene, a fluorescent isothiocyanate (FITC)-labeled rat anti-mouse CD3 monoclonal antibody (BD P harminge η ) was used as a T cell marker gene for detection in the dark spot. Dye for 30 minutes at 4 °C. After washing, the lymphocyte subtypes of each group were analyzed using a FACS Calibur (Becton Dickinson). Further, in order to remove the influence of dead cells on the flow cytometry analysis, dead cells were stained with iodopropylidene (PI: 2 // g/ml; Sigma), and PI positive cells were excluded and analyzed. (4) Results The results of flow cytometry analysis are shown in Table 1. It was judged that the anti-CD2 2 antibody screened by the present invention did not have much influence on the survival of B cells. Therefore, it will not be -37-200920399 to make the important B cells disappear in the body defense, and it is not expected to induce serious infection. [Table 1] B cell ratio (%) T cell ratio (%) Normal animal 11 87 EAE model 25 73 Anti-CD22 antibody administration group 21 77 Example 8: Anti-mouse CD22 antibody to natural mice Effect of the erythematosus (S LE ) model The SLE model mice were administered with the mouse CD 2 2 antibody C having the IL-6 production-inhibiting gl shown in Example 5, and the therapeutic effect was examined using the urinary occult blood fraction as an index. (1) Natural-type mouse S L E model NZW X BXSB F1 mice (Sakamoto SLC) using SLE-like pathological natural disease were used. Feeding to the 8th week of morbidity for the experiment. This SLE-like pathology is caused by b 1 cells that are as ill-conditioned as human SLE (Autoimmunity 1 5, 99- 1 05 (1 993 )) ° (2) Urinary occult blood (〇 ccuitblood) scores take fresh urine every 2 weeks. Use the Ames urine test strip (Hema-combistix 'Bayer') to check the occult blood for the occult blood score. (3) Method of administration The SLE model animal was administered with a dose of 2 〇〇 # g/head, and at 8 weeks of age, once a day, within 5 days, the venous injection of Example 5 was carried out by the tail vein. - 200920399 Anti-mouse c D 2 2 antibody C with inhibitory effect of IL-6 (N: The above antibody was diluted with physiological saline. (4) Results SLE-like pathology after administration of anti-mouse CD22 antibody Figure 5. Anti-mouse antibody administration group showed progression of lupus nephritis with significant early onset compared to control group. The inhibitory effect lasted for 3 months or longer. Example 9: Anti-mouse CD22 antibody against small Effect of the murine type II gel eyebrow inflammation (CIA) model As shown in Example 5, CIA model mice were administered anti-mouse c D 2 2 with biosuppressive ability and IL-10 inducing ability to joint The inflammation score is used as an indicator to explore its therapeutic effect.

(1) mouse π-type collagen-induced arthritis (CIA) model;; CIA of the animal model of chronic rheumatoid arthritis (RA) uses the same B-news activation and self-study as human chronic rheumatoid arthritis Mouse Fc 7 Rllb deficient mice produced by body antibody: Exp. Med. 1 46:8 5 7-8 6 8 (1 977)) ° Specifically, F-deficient C57BL/6 mice (10 weeks old; Female) subcutaneously-administered collagen (collagen technical begging) and complete adjuvant H3 7RA (the suspension was immunized once to induce CIA. (2) Arthritis score was based on the following criteria, and the score was 0 to 4 points per The extremity judgment of the mice is such that the arthritis score of the mouse is the sum of the limbs (maximum = 10). The 〇i degree 7F is suppressed in the depression - the end of the i-induced IL-6 production antibody B 匕 construction model is confirmed to be excessively performed (J c 7 Rllb to cattle II D i fc ο ) 重 severe illness: 16 points -39- 200920399 ) (Int Arch Allergy Appl Immunol. 35:456-467 (1 969)) 0 criterion: 〇 point: asymptomatic; Point: only one of the small joints of the limbs is swollen and red; 2 points: more than 2 small joints or larger joints such as hand or foot. ; 3 points: 1 hand or foot all red and swollen; 4 points: further 1 hand or foot swelling to the maximum (3) antibody administration method for the CIA model animal, the dose of 200 β g / head The anti-mouse CD22 antibody B or the anti-mouse IL-6 receptor (il-6R) antibody (Clone D7715A7) shown in Example 5 was administered intraperitoneally once a day from the immunization day (day 0). , BD Pharmingen) ( N=1 〇). The above antibody was diluted with physiological saline and used. (4) Results The progress of arthritis after administration of anti-mouse CD22 antibody is shown in Fig. 6. The anti-mouse CD22 antibody B significantly inhibited the progression of arthritis with respect to the control group. This inhibition effect continued during the observation period until 2 months after the end of the administration.

Since the anti-mouse CD22 antibody B obtained by the present invention was judged to strongly inhibit IL-6 production from the results of Example 5, it was effective against the IL-6R antibody which blocked the IL-6 signal. As a result, the anti-CD22 antibody B showed a significant sustained effect over the IL-6R antibody. Example 1 〇: Screening system for anti-human CD22 using human peripheral blood mononuclear cells (PBMC)-40- 200920399 (1) Blood collection method as an anticoagulant at the time of blood collection, using heparin sodium (Mitsubishi Well Pharma; final concentration 50 units) or EDTA (GIBCO; final concentration 5 mM), blood was collected from healthy adult volunteers. (2) Separation of peripheral blood mononuclear cells (pb MC) was performed using Lymphoprep (AXIS-SHIELD), a blood single-pass PBMC from a healthy adult volunteer according to the general method (Sc and. J. Clin Lab·Invest, 21 , Suppl. 97). The resulting PBMC was prepared in a culture medium to a specific cell concentration. (3) Culture method PBMC was inoculated into a round bottom 96-well plate at 1 〇 5 cells/well, and after adding various mouse anti-human CD22 antibody for several hours, an anti-human IgM F of 1 〇" g/mL was added. (ab') 2, stimulate cells. After culturing for 48 hours at 37 ° C in a CO 2 incubator, the culture supernatant was centrifuged and recovered. (4) Method for measuring IL-6 production and IL-10 production The culture supernatant was appropriately diluted to correspond to the IL-6 production amount and the IL-10 production amount of the ELISA kit (human IL-6 BD). OptEIA ELISA set, human IL-1 0 BD OptEIA ELI SA set, BD Biosciences) assay. (5) Screening method for anti-human CD22 antibody The mouse anti-human C D 2 2 antibody obtained in Example 4 was evaluated for IL-6 production inhibitory activity and IL-10 production-inducing activity. That is, various mouse anti-human CD22 antibodies were added to PBMC, and after about 6 hours of culture, add -41 - 200920399 and add anti-human IgM F(ab,) 2 at a concentration of #g/mL, and cultured in C02. Incubate at 7 7 ° C for 48 hours. The amount of IL-10 produced by IL-6 in the supernatant was determined. (6) Results As shown in Fig. 7, an anti-human C D 2 2 antibody which showed an inhibitory activity against IL-6 production by 50% compared with the control group was identified. Further, as shown in Fig. 8', an anti-human C D 2 2 antibody which showed an activity of more than 200% higher than IL-1 of the control group was identified. Example 1 1 : Correlation between Antigenicity of Anti-Human CD22 Antibody and Production Inhibitory Activity The anti-binding group of mouse anti-human CD22 antibody obtained in Example 4 was present in the region of the human CD 2 2 extracellular region, and further Its IL-6 produces an inhibitory activity, and explores the correlation between the inhibitory activity of the epitope-determining IL-6 production. (1) The epitope of mouse anti-human CD22 antibody exists. The G11-14 showing the human CD 2 2 extracellular domain 1 to 7 obtained in Example 3, the B4-152 of the expression region 3 to 7, and the region 5 were examined. To the B9-4 of 7 and the NSO cells of the host cell, the mouse anti-human CD 2 2 antibody binding site obtained by the screening of the present invention obtained in Example 4 was determined by cytometry analysis. That is, the three kinds of human CD22 expression cells and the NS0 cell suspension of the host cells were firstly distributed, and the amount of the mouse anti-human CD 2 2 antibody box of Example 4 was respectively induced to induce IL-6. After the suspension and mixing of the probe and the positional expression flow method -42- 200920399, after culturing for 30 minutes at 4 ° C in the dark, wash it. Subsequently, FITC-labeled goat anti-mouse IgG antibody F(ab')2 (Immunobiological Research Institute) was added as a secondary antibody, and it was washed in the dark at 30 ° C for 30 minutes. These samples were analyzed using FACS Calibur (Becton Dickinson) to determine the regions in which various mouse anti-human CD22 antibody epitopes were present. Further, in order to remove the influence of dead cells on the flow cytometry analysis, dead cells were stained with PI (2 // g/ml; Sigma), and PI positive cells were excluded and analyzed. As a result, an antibody that binds to G1 1-14 but does not bind to B4-152 and B9-4, that is, an antibody that binds to regions 1 and 2, is considered to have four 'and G 1 1 - 1 4 and B. 4 -1 5 2 antibodies that bind to B 9 -4, that is, 4 antibodies that bind to regions 3 and 4, and G1 1-14, B4-l52, and B9-, respectively. 4 bound antibodies, that is, 4 antibodies which are considered to bind to regions 5 and 7. (2) Inhibition of IL-6 production by anti-human CD22 antibody The 12 antibodies obtained in the above (1) were screened by the method described in Example 10 to investigate the IL-6 production inhibitory activity. (3) Results The results are shown in Fig. 9. The mouse anti-human C D 2 2 antibody combined with the regions 3 and 4 showed strong IL-6 production inhibitory activity. Example 2: Acquisition of full human type anti-human CD22 antibody and screening thereof -43- 200920399 (1) Antigen The two types of recombinant human CD22 highly expressed NS 0 cells obtained in Example 3 were used as fully human type anti-human CD22 antibody. The antigen is produced, that is, in addition to G 1 1 - 14 (a highly expressed colony of the full-length human CD 22 extracellular region) and B4-152 (regions 2 and 2 of the mouse and regions 3 to 7 of the human) In addition to the highly-selected strains of the hybrid type CD22, the following two types of antigens are also used. Human CD22 extracellular domain protein: In order to obtain human CD22 extracellular domain protein, human CD22 cDNA (Open Biosystem Inc) was used to construct a full-length human CD22/3 with a hexaminic acid tag at the C-terminus. Expression vector encoded by the extracellular domain protein (CD22ECD). Subsequently, transfection was carried out in Chinese hamster ovary cell line CHO cells, and stable and highly expressed transformants were selected. CD22ECD was purified from its culture supernatant by metal chelate affinity chromatography. High-expression CHO cells using a full-length human CD 22 extracellular region: Human CD22 cDNA (Open Biosystems Inc) and PCR primers (SEQ ID NOs: 15 and 16) were used to construct a expression vector encoding the full-length human CD22. This expression vector was then transfected into CHO cells to obtain a transformant. Next, the CHO cells highly expressed against the full-length human CD22 were isolated by flow cytometry based on the reactivity of the fluorescently labeled anti-human C D 2 2 antibody (Becton Dickinson Pharmingen, the selected strain Lu-HCL-1). (2) Human antibody-producing mouse system The mouse used in the production of a fully human type anti-human CD22 antibody -44- 200920399 The system is as follows. HuMAb-Mouse (registered trademark) 3 system (HCol 2 > HC〇20 'HCol 2/Balbc), followed by KM-Mouse (registered trademark) and KM- and HAC systems, all of which have human antibody genes Site. In these mouse systems, the endogenous mouse antibody light chain/c gene locus is inactivated (Chen et al. (1 993) EMBO J. 1 2:8 1 1 -820 ) and

The endogenous mouse antibody heavy chain gene locus is inactivated (International Publication WO 0 1 /09 1 87). On the other hand, these mouse systems maintain the human chromosome 14 fragment SC20 (International Publication WO 0 2/43 478), which maintains the human antibody light chain/C gene locus KCo5 (Fishwild et al) (1 996) Nature Biotechnology 1 4 : 8 4 5 - 8 5 1 ) and human antibody heavy chain gene loci. The HCo 1 2 system maintains the human antibody heavy chain gene locus transgene HCol2 (International Publication WO 0 1 /09 1 87). The HC(R) 20 system was produced as described in Example 4 of U.S. Provisional Application Serial No. 60/995,194. The details are as follows. The HC〇20 system was simultaneously introduced into a system comprising a transgenic PHC2 of an antibody heavy chain variable region, pVx6 of a heavy chain variable region, and YAC ylghlO comprising an antibody heavy chain variable region of a human embryonic strain. PHC2 alone has the ability to completely re-encode human immunoglobulin heavy chain variable region genes in vivo. The pVX6 and ylghl 0 additions are intended to help increase the diversity of the heavy chain variable regions of the embryonic plant. The individual components from the D N A mixture used in the preparation of H C 〇 20 are described below. pHC 2 has a human immunoglobulin heavy chain gene locus', that is, four heavy chain variable fragments derived from human embryonic strains 1-69 (-45 - 200920399 DP-10), 5-51 (DP-73), 4-34 (DP-63) and 3-30.3 (DP- 46). pHC2 in turn has 15 D fragments, 6 J fragments, ? The human genome sequence consisting of the 4 and yl normal regions and the P-Y1 switch region. PVx6 is a plastid ρ343·7.16 having VH1-18 (DP-14), a plastid p251f having ¥5_51 (DP-73), and having a heavy chain variable region (hereinafter referred to as VH) of a human embryo strain. The plastid pll2_2RR.7 of VH3-23 (DP-47) is constructed so as to have the VH regions of the three types. Ρ343.7·16 was introduced into the plastid vector PSP72 (Promega) and introduced 8.5 kb Hindlll/Sall fragment containing VH1-18 (DP-14) and 5' side group (2.5 kb) and 3' side base genome. Produced by sequence (5kb). P251f was inserted into the plastid selection vector pGFlf (Tayler et al. (1992) Nucleic Acids Res. 20:6287-6295) and simultaneously inserted a 7 kb BamHI/Hindllll fragment containing VH5-51 (DP-73) and 5, side The basal (5 kb) and 3' flanking genomic sequences (lkb) were made. P112.2RR.7 was inserted into the plastid selection vector pGFlk and inserted into the 10 kb EcoRV/BamHI fragment containing VH3-23 (DP-47) and the 5' side group (4 kb) and 3, the side group genomic sequence (5 kb). And making. Next, P251f cut the resulting 7 kb fragment with BamHI/Sall, cut it with BamHI/Sall and inserted P112.2RR.7 to obtain plastid pVx4. This pVx4 was cleaved by Xhol. At this position, the 8.5 kb fragment of P343.7.1 6 cut with Xhol/Sall was inserted in the same direction as the other two VH genes, and the plastid obtained finally was pVx6. The YAC ylgHIO system was initially mapped to human chromosome 14 using VH3 and VH4-specific primers by means of P C R screening. ylgHIO is a VH group and is established to contain VH1, VH2 -46-200920399, VH3 and VH4 (especially at least VHl-18, VH, 24, VH2-26, VH3-15, VH3-20, VH3-21, VH3-23) , VH3-30, VH3-30.5, VH3-30.3, VH3-33, VH4-30, VH4-30.4, VH4-30.3 > VH4-3 1, VH4-34). The pHC2 (80 kb) obtained as above, the PVx6 source fragment (26 kb) cut with Notl, and the ylgHIO (37 5 kb) were mixed at a molar ratio of 1:1:1, following the method of Hogan et al. (B. Hogan et al 1., Operation of Mouse Embryos, Laboratory Manual, 2nd Edition '1 994, published by Cold Spring Harbor Laboratory, Plainview, NY), microinjected to BDF1 x KCo5-CMD-JKD F1 embryo 1.5 years old In the nuclear. A system having the pVx6, pHC2, and ylgHIO source ratios obtained from the embryo was designated as HC〇20. HC〇20 system and CMD mutant mutant mice (International Publication No. 0 1 /09 1 8 7), JKD mutant mutant mice (Chen et al, (1 993) EMBO J. 12:81 820) KCo5 9272TG mice (Fishwild et al. (1 996) Nature Biotechnology 1 4: 845 -8 5 1 ) were crossed. The resulting mouse has human immunoglobulin heavy and light chain gene loci. The details of the KM-Mouse (registered trademark) system are described in U.S. Patent Application Publication No. 20020199213. The KM-λ HAC (HAC: Human Artificial Chromosome) system is quite similar to the ΚΜ-Μ 〇 u s e (registered trademark) system. The κ Μ - Μ 〇use (registered trademark) system introduces the KC 〇5 transgene and human 1 4 when the endogenous mouse antibody light chain/c gene locus and the mouse antibody heavy chain locus are inactivated. Chromosomal fragment SC20. In addition to these, the KM-AHAC system also introduces an artificial chromosome derived from human chromosome 22 with the human antibody light chain lambda locus. Therefore, if the ΚΜ-Λ HAC system is used, a human antibody having a human antibody light chain; I or /c can be produced. The KM-AHAC system is described in detail in U.S. Patent Application Publication No. 2 0 0 0 0 1 5 9 5 8 . (3) Acquisition of fully human type anti-human CD 22 antibody In order to produce a fully human type anti-human CD 22 antibody, the same procedure as in Example 2 and Example 4 was carried out. If briefly described, the antigen of Example 1 2 (1), that is, two types of recombinant human CD 2 2 high-expressing NS 0 cells (G11-14 and B4-152), and the full-length human CD22 extracellular region exhibit high expression of CHO cells, In the human CD22 extracellular domain protein, one of the antigens and the adjuvant (RIBI-Sigma # M65 3 6 ) were mixed in a 1:1 mixture to immunize the human antibody producing the various systems of Example 12 (2). 8 to 12 times. A method for producing a fully human type anti-human CD22 antibody using a human antibody is generally used in accordance with 1^〇!^6;^,1 to &1· (1 994) Nature 3 6 8 (6474): 8 5 6 - 8 5 9; Fishwild, D. et al. (1996) Nature Biotechnology 14: 845-851, International Publication WO 98/248 84. After measuring the antibody valence of the immunized mouse, the individual having the antibody valence was sufficiently increased by using the appropriate antigen of Example 1 2 (1) twice. Mouse spleen cells and mouse myeloma cell line P3X63-Ag 8.6 5 3 cells (ATCC, CRL 1581) were used, and PEG method (Nature 25 6:495-497, I975) or cell pulsed large-cavity cell fusion electroporation The instrument (Cyto Pulse Science, Inc., Glen Burnie, MD) was used for cell fusion using electrofusion based on the electric field. In the same manner as in the case of the second and fourth embodiments, the cells cultured with the HAT selected after the fusion -48-200920399 were used for screening, and the fully human-type anti-human CD22 monoclonal antibody-producing cells (hybrids) were selected. Next, 30 types of fully human-type anti-human CD22 monoclonal antibodies that specifically react with human CD22 were identified to produce hybridomas. For the analysis of the characteristics of the antibodies, 30 kinds of fully human type anti-human CD22 monoclonal antibodies were prepared from the culture supernatant obtained by culturing the hybrid tumor according to the general purification method (Current Protocols In Immunology Unit 2.6-2.7). These antibodies are referred to as "complete human type anti-human CD22 monoclonal antibodies". (4) Inhibition activity of IL-6 production by fully human type anti-human CD22 antibody The 30 fully human type anti-human CD22 antibodies obtained in the above (3) were screened by the method described in Example 1 to investigate IL- 6 produces inhibitory activity. (5) Results The results are shown in Figure 1. A fully human type anti-human CD22 antibody (20F4, 13B11, 23C6, 17H5, 12F6 and 1B1 1 ) showing an IL-6 production inhibitory activity of 40% or more compared with the control antibody can be identified. Example 13: Epitope analysis of fully human type anti-human CD22 antibody (1) Recombinant cells and host cells expressing various human CD22 extracellular regions are antigens of the fully human type anti-human CD22 antibody determined in Example 12. It is determined which of the human CD 2 2 extracellular regions is present in the region of -49-200920399, using cells expressing the following human CD22 extracellular regions. First, the three types of human CD22 highly expressed NSO cells obtained in Example 3 were used (G1 1-14: a highly expressed colony of the full-length human CD22 extracellular region, B 4 -1 5 2 : region 1 of the mouse) 2, and the hybrid CD22 high-performance selection strain of human region 3 to 7, followed by B9-4: the high-performance selection strain of hybrid CD22 in the region 1, 2 and the human region 5 to 7 of the mouse) As well as the NS0 cells of the host cell, explore the region where the epitope is present. Next, in addition to the high-expression CHO cells in the full-length human CD 2 2 extracellular region obtained in Example 1 2 (1), CHO cells and host CHO cells expressing various types of human CD22 extracellular regions prepared by the following methods were used. Further, the region in which the epitope is present is confirmed. Using human CD22 regions 1, 2, CHO cells: human CD22 cDNA (Open Biosystems Inc) and PCR primers (SEQ ID NOs: 17 and 18) were used to obtain DNA fragments of human CD22 regions 1, 2. Next, the human CD22 region 1, 2, the osteosporin (讯steonectin) signal sequence, and the rhodopsin transmembrane sequence are combined to construct an expression vector. This expression vector was transfected into CHO cells to obtain a transformant. Next, using the flow cytometry, the reactivity of the anti-human CD22 antibody (Becton Dickinson-Pharmingen, S-HCL-1) specific for the region 1 and 2 was detected by the flow cytometry, and the CD22 region was isolated from the human. 1, 2 high performance CHO cells. Mouse, region 1, 2, and human region 3 - 7 hybrid type c D 2 2 expressed C Ο Ο cells: -50- 200920399 The human CD22 extracellular region was expressed in regions 3-7, and the first mouse was produced as follows Human hybrid CD22 exhibits CHO cells. The mouse CD2 cDNA line was amplified by PCR using a mouse spleen cDNA gene library (BioChain Institute, Inc.) and a PCR primer (SEQ ID NOs: 19 and 20). Therefore, mhCD22-D3 is a DNA fragment produced by linking mouse CD22 regions 1 and 2 to regions 3 to C of human CD22. The chimeric DNA was produced by using a PCR primer (SEQ ID NOs: 16 and 21) to which a BstZ17I restriction enzyme cleavage site present at the end of the region 2 of the mouse CD22 was added, and using the amplified human CD22 DNA fragment. A performance vector containing mhCD22-D3 was produced, transfected into CHO cells to obtain a transformant, and the appropriate fluorescent labeling anti-human CD 2 based on the specific region 3 to 7 identified in the Example 藉 by the flow cytometry method. 2 The reactivity of the antibody is independent of the high-performance C Η Ο cells. This C D 2 2 shows that C Ο cells can express hybrid CD22 in regions 1, 2 of the mouse and regions 3 to 7 in humans. Mouse, region 1, 2 and human. Region 5-7 hybrid CD22 expresses C Ο Ο cells: the region 5-7 of the human CD 2 2 extracellular region is expressed as follows: Preparation of second mouse·human hybrid CD22 expression CHO cells. mhCD22-D5 is a D N A fragment produced by linking mouse CD22 regions 1 and 2 to regions 5 to C of human CD22 using the aforementioned mouse C D 2 2 c D N A . The chimeric DNA was amplified using the PCR primer (SEQ ID NO: 16 and 22) of the restriction enzyme cleavage site present at the end of the region 2 of the mouse CD22 using the amplified human CD22 DNA fragment-51 - 200920399 Production. Making a expression vector containing mhCD22-D5, transfected into CHO cells to obtain a transformant' by the flow cytometry method, based on the appropriate fluorescently labeled anti-human CD identified in the specific region 5 to 7 in Example 11. 2 2 Reactivity of antibodies, single high-performance CHO cells. This CD22 expression shows that CHO cells can express heterozygous CD22 in regions 1, 2 of the mouse and regions 5 to 7 in humans. (2) Method Among the antibodies screened in Example 12, 6 kinds of fully human type anti-human0022 antibodies (20?4, 13B11, 23C6, 17H5, 12F6 and 1B11) having IL-6 production inhibitory activity of 40% or more were selected. ), which region of the human CD 2 2 extracellular region is present in which the epitope is determined. That is, the three types of human CD22 obtained in Example 3 were respectively expressed as NSO cells and NSO cells of the host thereof, (1) of Example 12, and the human C D 2 2 obtained by (1) of Example 13 (1). A suspension of CHO cells and CHO cells of the host thereof was mixed with 6 kinds of fully human type anti-human CD22 antibodies, and washed in the dark at 15 ° C for 15 minutes, and then washed. Then add the pe standard as a secondary antibody. Several human IgGf/ij bodies F(ab')2 (Jackson ImmunoResearch Laboratories) were incubated in the dark at 30 °C for 30 minutes. After washing, the recognition area for 6 types of fully human type anti-human C D 2 2 antibodies was analyzed using FACS Calibur (Becton Dicknson). Further, in order to remove the influence of dead cells on the flow cytometry analysis, dead cells were stained with PI, and PI positive cells were excluded and analyzed. (3) Results -52- 200920399 The results are shown in Table 2. In the table, + indicates that the added antibody is combined - the antibody is not bound to the added antibody. [Table 2] _-Human CD22 region expression site antibody E SNS0| The cell expression is characterized by CHO fine and 1 S full-length region 3-7 region 5-7 control group full-length region 1 ' 2 region 3-7 region 5-7 Control group 20F4 + - - _ + . _ er 13B11 + - + + . . 17H5 + + + turn + + 1B11 + + - _ + + . 12F6 + + + - + _ + + 23C6 + + - - + _ + - It is understood that none of the antibodies bind to 1, 2 of the extracellular domain of human CD 2 2 . Example 14: B cell injury effect of a fully human type anti-human CD22 antibody A fully human type anti-human CD antibody which has been shown to exhibit IL-6 production inhibitory activity in Example 12 has been investigated for its B cell killing effect (1) Method The human B cell line Ramos (ATCC) was suspended in RPMI1640 medium containing 10% fetal calf serum, and seeded in a 96-well plate at 3xl05/200#L/well. -53- 200920399 Addition of Rituximab (anti-human CD20 antibody; human IgG1), chimeric LL2 (International Publication WO 20000747 18, anti-human CD22 antibody; human IgG1), trastuzumab (Trastuzumab) (anti-human HER2 antibody; human IgG1) or novel anti-human CD22 antibody 13B1 1, 17H5, 20F4 (all human IgG1) to a final concentration of 0.2 to 5 # g/mL. Further, a goat anti-human IgG Fc antibody (manufactured by Cappel Co., Ltd.) having a final concentration of 20 // g/mL was added, and cultured at 37 ° C for 2 days. The number of the cells was fluorescently stained with a BD cell survival kit (BD Biosciences), and then measured by a FACS Calibur (manufactured by Becton Dickinson Co., Ltd.). (2) Results The cell rate when the number of cells in the absence of antibody was taken as 100% is shown in Table 3. Trastuzumab in the negative control group had no effect on the number of B cells. On the other hand, the anti-C 2 2 antibody preparation used as a positive control group, the concentration of the chimeric LL2 of the rituximab and the anti-CD22 antibody, decreased the number of B cells. 5 // g/mL of rituximab or chimeric LL2 reduced the number of B cells by 30% and 59%, respectively. On the other hand, the fully human type anti-human CD22 antibody 13B1 1 , 1 7G5, 2 0F4 showing inhibition of IL-6 production in Example 12 has weak killing ability to B cells even at 5 // g/mL of living cells. The rate is also as high as 7 6~9 6 %. In other words, in the same manner as the autoimmune disease model of the seventh embodiment, the antibody identified by the screening system of the present invention does not cause a serious effect on the disappearance of B cells which are important for the defense of the living body. And useful. -54- 200920399 [Table 3] Antibody concentration 0.2 β g/mL 1 μg/mL 5 β g/mL No addition of 100% ± 14% 100% ± 14 ° / 〇 100% ± 14 ° / 〇 妥 珠 珠 单Anti-94% ± 5% 92% ± 14 〇 / 〇 8 8% ± 8 ° / rituximab 90% ± 10% 77% ± 24% 3 0% ± 2 ° / 〇 chimera LL2 90% ± 7% 75% ± 9% 5 9% ± 4 〇 / 〇 13B11 95% ± 5% 98% ± 18 ° / 〇 76% ± 5 ° / 〇 17H5 106% ± 2% 9 5% ± 9% 96% ± 13% 20F4 102%±14°/〇109%±16% 86%±40/〇Example 1 5: Complementarity of fully human type anti-human CD22 antibody determines region sequence analysis IL resolved in the above Example 13 - 6 produces a base of cDNA encoding heavy and light chain variable regions of 6 types of fully human type anti-human CD 22 antibodies (20F4, 13B11, 17H5, 1B11, 12F6 and 23C6) having an inhibitory activity of more than 40% The base sequence analysis and the protein chemical analysis of the expressed protein are carried out according to a general method. Sequence number 2 3 of the sequence listing and Figure 11 show the sequence of the heavy chain variable region of 1 B 1 1 . The heavy chain variable region of 1 B 1 1 is derived from human embryonic strains 3-21 and JH3b. The isotype of the normal region of the heavy bond is human IgG1. Sequence number 24 of the sequence listing and Figure 12 show the sequence of the light chain variable region of 丨b 1 1 . The light chain variable region of B 1 1 is derived from A27 and JK2 of human embryonic strains. The isotype of the normal region of the light chain is human κ. Sequence number 25 of the sequence listing and Figure 13 show the sequence of the heavy chain variable region of 12F6. The heavy chain variable region of 1 2 F 6 is derived from human embryonic strain -55-200920399 4-34, JK5b. The isotype of the heavy chain normal region is the sequence number of the sequence listing 2 6 and the sequence of the map variable region of Fig. 14. The light chain variable region of 1 2 F 6 is L6 and JK4. The isoform of the normal region of the light chain is the sequence number of the human sequence list 2 7 and the sequence of the variable region is shown in Fig. 15. 1 3 B 1 1 The heavy chain variable region is 3-2 3, JH4b. The isotype of the heavy chain normal region is the sequence number 2 8 of the sequence listing and the sequence of the variable region is shown in Fig. 16. 1 1 3 B 1 1 light chain variable region L15, JK1. Also, the homologous 正常 of the normal region of the light chain SEQ ID NO: 29 of the sequence listing and Figure 7 show the sequence of the variable region. 1 7 Η 5 heavy chain variable region system 3-3 0.3, JH4b. The sequence of the normal region of the heavy chain sequence number 3 0 and the sequence of the variable region are shown in Fig. 18. 1 7 Η 5 light chain variable area system Α 10, JK2. The isoform of the light chain normal region is 序列 sequence number 3 1 of the sequence listing and Figure 19 shows the sequence of the variable region. The heavy chain variable region of 20F4 is 4-59, JH2. Further, the isotype of the heavy chain normal region is the sequence number 3 2 of the y sequence table and the sequence of the variable region is shown in Fig. 20 . The light chain variable region of 20F4 is L15 and JK2. The isoform of the normal region of the light chain is the sequence number 3 3 of the human sequence listing and the sequence of the variable region is shown in Fig. 2 1 . The heavy chain variable region of 23C6 is human IgG1. ;: The light chain of 1 2 F 6 can be derived from human embryo strain /c. The heavy chain of 1 3B 1 1 can be derived from human embryonic strain human IgG1. The light chain of 1 3 B 1 1 can be derived from human embryos; human/c. The heavy chain of 1 7 Η 5 can be derived from human embryonic strains as human IgG1. The light chain of 17H5 can be derived from human embryonic strains, class /C. The heavy chain of 20F4 can be derived from human embryonic strains, IgG1. : The light chain of 20F4 can be derived from human embryonic strains. Class /c. The heavy chain of 23C6 can be derived from human embryonic strain -56-200920399 1-69, JH6b. The isotype of the heavy chain normal region is human IgG1. Sequence number 34 of the sequence listing and Figure 22 show the sequence of the light chain variable region 1 (V / c · 1 ) of 23 C6. The light chain variable region 1 of 2 3 C 6 is derived from L6 and JK1 of human embryonic plants. Further, the isoform of the normal region of the light chain is human K 序列 Sequence No. 3 5 of the Sequence Listing and Figure 2 3 shows the sequence of the light chain variable region 2 (V / c. 2) of 2 3 C 6 . The light chain variable region 2 of 2 3 C 6 is derived from L6 and JK1 of human embryonic plants. The fully human type antibody CD22 antibody 23C6 is disclosed in International Patent Application No. PCT/US2007/0 8 6 1 52, the details of which are as follows. By comparing the heavy chain antibody sequence of 23C6 with the heavy chain antibody sequence of a known human embryo strain, it was revealed that the 23C6 heavy chain system utilizes the VH region derived from human embryonic strain VH 1-69. The D region is derived from the human embryonic strain 2-15' JH region using human embryonic strain JH 6B. In order to determine the C D R region in the VH sequence of 23C6, the K a b a t system was used for analysis, and Figure 21 shows the CDR1, CDR2, and CDR3 regions. By comparing the 2 3 C 6 V / c . 1 light chain antibody sequence with the known human embryonic strain / C light chain antibody sequence, the 23C6V /c · 1 light chain shows that the V / C line is derived from human embryonic strain V / c L6, JK region is derived from human embryonic strain JK1. To determine the CDR regions in the V/c.1 sequence of 23C6, the Kabat system was used for analysis, and Figure 22 shows the CDR1, CDR2, and CDR3 regions. By comparing the 23 C6 V/c.2 light chain antibody sequence with the known human embryonic strain/C light chain antibody sequence, the 23C6V/c.2 light chain shows that the V/c line is derived from the human embryonic strain V/c L6. The JK region is derived from the human embryonic strain JK 1. -57-200920399 To determine the CDR regions in the V/c.2 sequence of 23C6, the Kabat system was used for analysis. Figure 23 shows the CDR1, CDR2, and CDR3 regions. [Industrial Applicability] According to the present invention, it is possible to provide an effective therapeutic and/or preventive agent for a disease selected from diseases associated with IL-6 and/or IL-10 and B cell lymphoid tumors. Further, a screening method for efficiently selecting antibodies effective for such diseases can be provided. Further, since the antibody of the present invention exerts an IL-6 production inhibitory action and/or an IL-10 production-enhancing effect without causing the B cell to disappear, it is considered to be clinically useful because it does not induce a serious infection. This application is based on Japanese Patent Application No. 2007-160531 (filed on Jun. 18, 2007), the entire contents of which are hereby incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic view showing the CD22 molecular portion of three recombinant human CD22-expressing cells obtained in Example 3. (A) shows the molecular pattern represented by the recombinant CD22 expressing cell line G1 1-14, and (B) shows the molecular pattern represented by the recombinant CD22 expressing cell lines 34-1/30 and B4-152. (C) shows a molecular pattern diagram expressed by recombinant CD22 expressing cell line B9-4. Fig. 2 is a graph showing the results of measurement of the inhibitory activity against IL-6 production by the anti-mouse CD22 antibody of Example 5. Fig. 3 is a graph showing the results of measurement of the induction activity of IL-10 produced by the anti-mouse CD22 antibody of Example 5. Fig. 4 is a graph showing the effect of the mouse model CD22 antibody against the mouse E A E model of Example 6, using survival rate as an index. The survival rate when anti-CD22 antibody A was administered, and (B) indicates the survival rate of CD22 antibody BB. Fig. 5 is a graph showing the effect of administering the anti-mouse CD22 antibody to the mouse model of whole-spotted lupus (SLE) of the natural disease of Example 8 (occult blood) as an index. Fig. 6 is a graph showing the effect of administering the anti-mouse CD22 antibody to the mouse type II collagen inflammatory (CIA) model of Example 9 with the number of points as an index. Fig. 7 is a graph showing the results of measurement of the inhibitory activity against mouse anti-human CD22 anti-IL-6 production of Example 10. The bar "0" indicates the amount of I L - 6 produced without stimulation. The column "anti-1 g Μ" indicates the amount of IL-6 produced by the addition of human IgM F(ab') 2 to stimulate PBMC. The photograph indicates the amount of IL-6 produced by the addition of anti-human IgM F(ab') 2 stimulation. The column "1 to 26" indicates the amount of IL-6 produced by various mouse anti-human CD22 antibodies (No.) added under anti-human F(ab')2 stimulation. Fig. 8 shows the mouse of Example 10. The results of the measurement of the inducing activity against human CD22 against IL-1 0. The column "〇" indicates the amount of IL-10 produced without stimulation. The column "anti-1 g M" indicates the addition of human IgM F (ab, 2) The amount of IL-10 produced by PBMC is stimulated. "Control" means that the anti-small (A) table is added to the anti-small (A)-stimulated anti-human IgM joint. Inflammatory splitting of PBMC plus anti-column "control antibody class IgM 1 ~26" body pair PBMC plus anti-column "Control anti-59- 200920399 body IL-10 production. Column "1~26 The expression of IL-10 produced by addition of various mouse anti-human CD22 antibodies (No. 1 to 26) stimulated by anti-human IgM F(ab') 2 . Fig. 9 shows the inhibitory activity of the mouse anti-human CD22 antibody which binds to each region of human CD22 of Example 11 to IL-6 production. "No stimulation" indicates that the IL-6 concentration in the supernatant of the PBMC was not stimulated by the anti-IgM antibody, and "IgM antibody stimulation" indicates that the IL-6 concentration in the supernatant of the PBMC stimulated with the anti-IgM antibody was also the control group. Bars 1 to 4 show the results of addition of mouse anti-human CD22 antibody binding to regions 1 to 2, and bars 5 to 8 show the results of addition of mouse anti-human CD22 antibody binding to regions 3 to 4, The 9 to 12 lines show the results of adding mouse anti-human CD 2 2 antibodies bound to regions 5 to 7. Figure 10 is a graph showing the ability of the fully human type anti-human CD22 antibody of Example 12 to inhibit IL-6 production. "No stimulation" showed the amount of IL-6 produced when no IgM antibody was added to stimulate PBMC, and the other bars showed the amount of IL-6 produced by the addition of IgM antibody to stimulate PBMC. "Anti-IgM antibody stimulation" showed the amount of IL-6 produced by the addition of anti-IgM antibody to stimulate PBMC. The "control antibody" shows the amount of IL-6 produced when an unrelated monoclonal antibody is added. "Positive mouse anti-CD22 antibody" showed the amount of IL-6 produced when the mouse anti-human C D 2 2 antibody having the inhibitory effect on IL-6 production obtained in Example 4 was added. Figure 10 (a) shows the results of adding the full human type anti-human CD22 antibody No. 1 to No. 10 obtained in Example 12, and Figure 1 (b) shows the addition of a fully human type anti-human CD22 antibody No. 1 to No. 2 Results of the nickname, Figure 10 (c) shows the results of adding full human type anti-human-60-200920399 CD22 antibody Nos. 21 to 30. Figure 11 shows the heavy chain variable region sequence of the fully human type anti-CD22 antibody 1B11 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 12 shows the light chain variable region sequence of the fully human type anti-CD22 antibody 1B1 1 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 13 shows the heavy chain variable region sequence of the fully human type anti-CD22 antibody 12F6 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 14 shows the light chain variable region sequence of the fully human type anti-CD22 antibody 12F6 of Example 15. The position of the C D R s sequence is shown in the figure. Figure 15 shows the heavy chain variable region sequence of the fully human type anti-CD22 antibody 13B11 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 16 shows the light chain variable region sequence of the fully human type anti-CD22 antibody 1 3 B 1 1 of Example 15. The position of the C D Rs sequence is shown in the figure. Figure 17 shows the heavy chain variable region sequence of the fully human type anti-CD22 antibody 17H5 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 18 shows the light chain variable region sequence of the fully human type anti-CD22 antibody 1 7H5 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 19 shows the heavy chain variable region sequence of the fully human type anti-CD22 antibody 20F4 of Example 15. The position of the C D R s sequence is shown in the figure. Figure 20 shows the light chain variable region sequence of the fully human type anti-CD22 antibody 20F4 of Example 15. The figure shows the position of the c D R s sequence. Figure 21 shows the heavy chain variable region sequence of the fully human type anti-CD22 antibody 23C6 of Example 15. The position of the CDRs sequence is shown in the figure. Figure 22 shows the light chain variable region 1 (Vk.I) sequence of the fully human type anti-CD22 antibody of Example 15 -61 - 200920399 2 3C6. The figure shows the CDRs sequence position. Figure 23 shows the light chain variable region 2 (Vk. 2) sequence of the fully human type anti-CD22 antibody 23C6 of Example 15. The figure shows the CDRs sequence position. -62- 200920399 Sequence Listing <110>Mitsubishi Tanabe Pharmaceutical Co., Ltd. <120> antibody screening method and antibody obtained therefrom <130> 091255 <150> JP 2007-160531 <151>2007118 <160&gt 35 <170> Patent Version 3.3 <210> 1 <211>30 <212> DNA <213>Manual<220><223> Based on Mouse CD22 Primer<220> Inventor: Okamoto, Tadao: Miura, Masami; King, David; Masood,

Asna <400> 1 gggaattcgc cacaccaccc ccaaccgtgg 30 <210〉 2 <211> 30 <212> DNA <213>manual<220><223>based on mouse CD22 primer <400> 2 tttctagatc Aagaacctga aacagcagct 30 <210> 3 <211> 2607 <212> DNA <213〉家鼷鼠<220><221> CDS <222>(1)·(2607)

<223> cDNA based on mouse CD22 <400> 3 atg cgc gtc cat tac ctg tgg etc etc ttg ate eta gga cat gtg get 48

Met Arg Vai His Tyr Leu Trp Leu Leu Leu lie Leu Gly His Val Ala 15 10 teg get egg tac age tea gca aat gat tgg acc gtt gac cat ccc caa 96

Ser Ala Arg Tyr Ser Ser Ala Asn Asp Trp Thr Val Asp His Pro Gin 20 25 30 acc etc ttt gee tgg gag gga gee tgc ate agg att cct tgc aag tac 144

Thr Leu Phe Ala Trp Glu Gly Ala Cys Me Arg lie Pro Cys Lys Tyr 35 40 45 aaa act cca eta ccc aag gca cgt ctg gac aac ate etc ett ttt cag 192

Lys Thr Pro Leu Pro Lys Ala Arg Leu Asp Asn lie Leu Leu Phe Gin 50 55 60 aac tat gag ttt gac aag gee acc aag aaa ttc aca gga act gtc ctg 240 200920399

Asn Tyr Glu Phe Asp Lys Ala Thr Lys Lys Phe Thr Gly Thr Val Leu 65 70 75 80 tac aac gcc aca aag act gag aag gac cca gag tct gag ctg tac ctt 288

Tyr Asn Ala Thr Lys Thr Glu Lys Asp Pro Glu Ser Glu Leu Tyr Leu 85 90 95 tct aag caa ggg aga gta aca ttt ctg ggg aac aga ata gac aat tgt 336

Ser Lys Gin Gly Arg Val Thr Phe Leu Gly Asn Arg lie Asp Asn Cys 100 105 110 acc ctg aaa ate cac ccg ata cgt gcc aat gac agt ggg aat ctg ggg 384

Thr Leu Lys Ile His Pro Ile Arg Ala Asn Asp Ser Gly Asn Leu Gly 115 120 125 ttg agg atg acc gca ggg act gaa ega tgg atg gag ccc att cac etc 432

Leu Arg Met Thr Ala Gly Thr Glu Arg Trp Met Giu Pro He His Leu 130 135 140 aat gtc teg gag aaa ccg ttc caa cct tac ate cag atg cca tea gaa 480

Asn Val Ser Glu Lys Pro Phe Gin Pro Tyr lie Gin Met Pro Ser Glu 145 150 155 160 ate egg gaa tcc caa agt gtc act ctg acc tgt gga ctg aat ttc tcc 528 lie Arg Glu Ser Gin Ser Val Thr Leu Thr Cys Gly Leu Asn Phe Ser 165 170 175 tgc ttt ggg tat gac ate etc ttg aag tgg ttc eta gaa gat tct gag 576

Cys Phe Gly Tyr Asp Me Leu Leu Lys Trp Phe Leu Glu Asp Ser Glu 180 185 190 ate acc tcc ate acc tct tct gtc acc tcc ate acc tct tct gtc acc 624 lie Thr Ser lie Thr Ser Ser Val Thr Ser lie Thr Ser Ser Val Thr 195 200 205 tcc tcc att aag aat gtc tat aca gag age aag etc acc ttc caa cca 672

Ser Ser lie Lys Asn Vai Tyr Thr Glu Ser Lys Leu Thr Phe Gin Pro 210 215 220 aag tgg aeg gac cac gga aag agt gtg aag tgc caa gtc cag cac tcc 720

Lys Trp Thr Asp His Gly Lys Ser Val Lys Cys Gin Val Gin His Ser 225 230 235 240 tcc aaa gtg etc tea gag tgc act gtg cat ctg gat gtt aag tat acc 768

Ser Lys Val Leu Ser Glu Cys Thr Val His Leu Asp Val Lys Tyr Thr 245 250 255 ccg aag ctg gag ate aag gtc aat ccc aca gag gtg gaa aag aac aat 816

Pro Lys Leu Glu Ile Lys Val Asn Pro Thr Giu Val Glu Lys Asn Asn 260 265 270 tct gtg acc atg aca tgc egg gtt aac age age aac ccg aaa etc agg 864

Ser Val Thr Met Thr Cys Arg Val Asn Ser Ser Asn Pro Lys Leu Arg 275 280 285 acc gtg geg gtg tct tgg ttc aag gat ggg ege ccc eta gag gat cag 912

Thr Val Ala Val Ser Trp Phe Lys Asp Gly Arg Pro Leu Glu Asp Gin 290 295 300 gaa ctg gaa cag gaa caa cag atg tcc aag eta att ctg cat tea gtg 960

Glu Leu Glu Gin Glu Gin Gin Met Ser Lys Leu lie Leu His Ser Val 305 310 315 320 acc aag gac atg aga ggg aaa tac egg tgc cag get tcc aac gac ata 1008

Thr Lys Asp Met Arg Gly Lys Tyr Arg Cys Gin Ala Ser Asn Asp Me 325 330 335 ggc cca gga gag teg gaa gaa gtg gaa etc ag gtg cac tat get cca 1056

Gly Pro Gly Glu Ser Glu Glu Val Glu Leu Thr Val His Tyr Ala Pro 340 345 350 gaa ccc tcc agg gtt cac ate tac cct tcc ccc get gaa gag gga cag 1104

Glu Pro Ser Arg Val His Ile Tyr Pro Ser Pro Ala Glu Glu Gly Gin 355 360 365 -2- 200920399 tea gta gag ctg att tgt gag tea ctg gee agt cca agt gca aca aac 1152

Ser Val Glu Leu lie Cys Glu Ser Leu Ala Ser Pro Ser Ala Thr A$n 370 375 380 tac acc tgg tat cac aac agg aaa cct ata cct gga gac acc caa gag 1200

Tyr Thr Trp Tyr His Asn Arg Lys Pro lie Pro Gly Asp Thr Gin Glu 385 390 395 400 aag etc ege ate cct aaa gtc tee ccg tgg cat get ggg aat tac tet 1248

Lys Leu Arg lie Pro Lys Val Ser Pro Trp His Ala Gly Asn Tyr Ser 405 410 415 tgc ttg gca gag aac cgt ctg ggt cat gga aag ata gac cag gaa get 1296

Cys Leu Ala Glu Asn Arg Leu Gly His Gly Lys lie Asp Gin Glu Ala 420 425 430 aag ctg gat gtc cat tat get ccc aag geg gtg acc aca gtg att cag 1344

Lys Leu Asp Val His Tyr Ala Pro Lys Ala Val Thr Thr Val lie Gin 435 440 445 age ttc aca cca ate ctg gaa gga gac agt gtg acc ctg gtc tgt agg 1392

Ser Phe Thr Pro lie Leu Glu Gly Asp Ser Val Thr Leu Val Cys Arg 450 455 460 tac aac tee age aat cca gac gtc acc tee tac aga tgg aac cct caa 1440

Tyr Asn Ser Ser Asn Pro Asp Val Thr Ser Tyr Arg Trp Asn Pro Gin 465 470 475 480 ggt tet ggg agt gtg etc aaa ccc gga gtg ctg agg att cag aaa gtg 1488

Gly Ser Gly Ser Val Leu Lys Pro Gly Val Leu Arg lie Gin Lys Val 485 490 495 aca tgg gat tee atg cct gtc age tgt get gee tgc aac cac aag tgt 1536

Thr Trp Asp Ser Met Pro Val Ser Cys Ala Ala Cys Asn His Lys Cys 500 505 510 teg tgg gee etc cct gtc ate ctg aat gtc cac tac gee ccc aga gac 1584

Ser Trp Ala Leu Pro Val lie Leu Asn Val His Tyr Ala Pro Arg Asp 515 520 525 gtg aag gta ctg aag gta age ccc gca tea gag ate ege get ggg cag 1632

Val Lys Val Leu Lys Val Ser Pro Ala Ser Glu Me Arg Ala Gly Gin 530 535 540 cgt gtc etc etc caa tgc gac ttc gca gag age aac ccg gca gag gtc 1680

Arg Val Leu Leu Gin Cys Asp Phe Ala Glu Ser Asn Pro Ala Glu Val 545 550 555 560 ege ttc ttc tgg aag aag aat ggg agt etc gtg cag gaa ggg agg tac 1728

Arg Phe Phe Trp Lys Lys Asn Gly Ser Leu Val 6ln Glu Gly Arg Tyr 565 570 575 ctg age ttc ggc tee gtc tee cca gaa gat tet gga aat tat aac tgc 1776

Leu Ser Phe Gly Ser Val Ser Pro Glu Asp Ser Gly Asn Tyr Asn Cys 580 585 590 atg gtc aac aac tee ate gga gag acc ttg tea cag gee tgg aac etc 1824

Met Val Asn Asn Ser lie Gly Glu Thr Leu Ser Gin Ala Trp Asn Leu 595 600 605 caa gtg ctg tat get cct egg agg eta cgt gtg tee ate age cct ggg 1872

Gin Val Leu Tyr Ala Pro Arg Arg Leu Arg Val Ser lie Ser Pro Gly 610 615 620 gac cat gtg atg gag ggg aag aag gee acc ttg tee tgt gag agt gat 1920

Asp His Val Met Glu Giy Lys Lys Ala Thr Leu Ser Cys Glu Ser Asp 625 630 635 640 gee aat ccg ccc ate tea cag tac acc tgg ttt gac tee agt ggc caa 1968

Ala Asn Pro Pro lie Ser Gin Tyr Thr Trp Phe Asp Ser Ser Gly Gin 645 650 655 gac etc cac tee tea ggc cag aaa ctg aga ctg gaa ccc ctg gag gtc 2016 -3- 200920399

Asp Leu His Ser Ser Gly Gin Lys Leu Arg Leu Glu Pro Leu Glu Val 660 665 670 caa cac acg ggt tcc tac cgc tgc aaa ggg acc aat ggg ata ggc aca

Gin His Thr Gly Ser Tyr Arg Cys Lys Gly Thr Asn Gly lie Gly Thr 675 680 685 gga gag tea cca ccc age acc etc act gtc tac tac agt cca gag acc

Gi y y y y y y y y y y y y Phe 705 710 715 720 ate ctg gcc ate tgg ggg atg aaa ate cag aaa aaa tgg aag caa aac

Me Leu Ala Me Trp Gly Met Lys He Gin Lys Lys Trp Lys Gin Asn 725 730 735 cgc age cag cag ggg ett cag gaa aat tcc agt ggc cag age ttt ttt

Arg Ser Gin Gin Gin Leu Gin Glu Asn Ser Ser Gly Gin Ser Phe Phe 740 745 750 gtg agg aac aaa aag get agg agg acc cct etc tea gaa ggc ccc caa

Val Arg Asn Lys Lys Ala Arg Arg Thr Pro Leu Ser Glu Gly Pro Gin 755 760 765 tcc cag gga tgc tac aat ccg gca atg gat gac act gtt agt tat gcc

Ser Gin Gly Cys Tyr Asn Pro Ala Met Asp Asp Thr Val Ser Tyr Ala 770 775 780 ate ttg cgc ttt cca gag agt gac acg cac aat get gga gat gca ggg

Ile Leu Arg Phe Pro Glu Ser Asp Thr His Asn Ala Gly Asp Ala Gly 785 790 795 800 acc cca gca aca cag get cct cct cca aac aac age gac acg gtc act

Thr Pro Ala Thr Gin Ala Pro Pro Pro Asn Asn Ser Asp Thr Val Thr 805 810 815 tac teg gtg ata cag aag egg cct atg ggg gat tat gag aat gtg aat

Tyr Ser Val lie Gin Lys Arg Pro Met Gly Asp Tyr Glu Asn Val Asn 820 825 830 ccg age tgc ccg gag gat gag age ate cat tac tea gag ctg gtt cag

Pro Ser Cys Pro Glu Asp Glu Ser lie His Tyr Ser Glu Leu Val Gin 835 840 845 ttt ggg get ggt aag egg ccc cag gca aag gaa gac gta gac tat gtg

Phe Gly Ala Gly Lys Arg Pro Gin Ala Lys Glu Asp Val Asp Tyr Vai 850 855 860 acc etc aag cac tga Thr Leu Lys His 865 &lt;210&gt; 4 <211> 868 &lt;212> PRT &lt;213〉 Home Mole &lt;220> &lt;223> murine CD22 amino acid sequence &lt;400> 4

Met Arg Val His Tyr Leu Trp Leu Leu Leu lie Leu Gly His Val Ala 15 10 15

Ser Ala Arg Tyr Ser Ser Ala Asn Asp Trp Thr Val Asp His Pro Gin 20 25 30 2064 2112 2160 2208 2256 2304 2352 2400 2448 2496 2544 2592 2607 -4- 200920399

Thr Leu Phe Ala Trp Glu Gly Ala Cys lie Arg lie Pro Cys Lys Tyr 35 40 45

Lys Thr Pro Leu Pro Lys Ala Arg Leu Asp Asn lie Leu Leu Phe Gin 50 55 60

Asn Tyr Glu Phe Asp Lys Ala Thr Lys Lys Phe Thr Gly Thr Val Leu 65 70 75 80

Tyr Asn Ala Thr Lys Thr Glu Lys Asp Pro Glu Ser Glu Leu Tyr Leu 85 90 95

Ser Lys Gin Gly Arg Val Thr Phe Leu Gly Asn Arg Ile Asp Asn Gys 100 105 110

Thr Leu Lys lie His Pro lie Arg Ala Asn Asp Ser Gly Asn Leu Gly 115 120 125

Leu Arg Met Thr Ala Gly Thr Glu Arg Trp Met Glu Pro He His Leu 130 135 140

Asn Val Ser Glu Lys Pro Phe Gin Pro Tyr lie Gin Met Pro Ser Glu 145 150 155 160 lie Arg Glu Ser Gin Ser Val Thr Leu Thr Cys Gly Leu Asn Phe Ser 165 170 175

Cys Phe Gly Tyr Asp Ile Leu Leu Lys Trp Phe Leu Giu Asp Ser Glu 180 185 190 lie Thr Ser lie Thr Ser Ser Val Thr Ser lie Thr Ser Ser Val Thr 195 200 205

Ser Ser lie Lys Asn Val Tyr Thr Glu Ser Lys Leu Thr Phe Gin Pro 210 215 220

Lys Trp Thr Asp His Gly Lys Ser Val Lys Cys Gin Val Gin His Ser 225 230 235 240

Ser Lys Val Leu Ser Glu Cys Thr Val His Leu Asp Val Lys Tyr Thr 245 250 255

Pro Lys Leu Glu lie Lys Val Asn Pro Thr Glu Val Glu Lys Asn Asn 260 265 270

Ser Val Thr Met Thr Cys Arg Val Asn Ser Ser Asn Pro Lys Leu Arg 275 280 285

Thr Val Ala Val Ser Trp Phe Lys Asp Gly Arg Pro Leu Glu Asp Gin 290 295 300

Glu Leu Glu Gin Glu Gin Gin Met Ser Lys Leu lie Leu His Ser Val 305 310 315 320

Thr Lys Asp Met Arg Gly Lys Tyr Arg Cys Gin Ala Ser Asn Asp He 325 330 335 -5- 200920399

Gly Pro Giy Qlu Ser Glu Glu Val Glu Leu Thr Val His Tyr Ala Pro 340 345 350

Glu Pro Ser Arg Val His lie Tyr Pro Ser Pro Ala Glu Glu Gly Gin 355 360 365

Ser Val Glu Leu lie Cys Glu Ser Leu Ala Ser Pro Ser Ala Thr Asn 370 375 380

Tyr Thr Trp Tyr His Asn Arg Lys Pro lie Pro Gly Asp Thr Gin Glu 385 390 395 400

Lys Leu Arg lie Pro Lys Val Ser Pro Trp His Ala Gly Asn Tyr Ser 405 410 415

Cys Leu Ala Glu Asn Arg Leu Gly His Gly Lys lie Asp Gin Glu Ala 420 425 430

Lys Leu Asp Vat His Tyr Ala Pro Lys Ala Val Thr Thr Val lie Gin 435 440 445

Ser Phe Thr Pro Ile Leu Glu Gly Asp Ser Val Thr Leu Val Cys Arg 450 455 460

Tyr Asn Ser Ser Asn Pro Asp Val Thr Ser Tyr Arg Trp Asn Pro Gin 465 470 475 480

Gly Ser Gly Ser Val Leu Lys Pro Gly Val Leu Arg lie Gin Lys Val 485 490 495

Thr Trp Asp Ser Met Pro Val Ser Cys Ala Ala Cys Asn His Lys Cys 500 505 510

Ser Trp Ala Leu Pro Val lie Leu Asn Val His Tyr Ala Pro Arg Asp 515 520 525

Val Lys Val Leu Lys Val Ser Pro Ala Ser Glu Ile Arg Ala Gly Gin 530 535 540

Arg Val Leu Leu Gin Cys Asp Phe Ala Glu Ser Asn Pro Ala Glu Val 545 550 555 560

Arg Phe Phe Trp Lys Lys Asn Gly Ser Leu Val Gin Glu Gly Arg Tyr 565 570 575

Leu Ser Phe Gly Ser Vai Ser Pro Glu Asp Ser Gly Asn Tyr Asn Cys 580 585 590

Met Vai Asn Asn Ser lie Gly Qlu Thr Leu Ser Gin Ala Trp Asn Leu 595 600 605

Gin Val Leu Tyr Ala Pro Arg Arg Leu Arg Val Ser Ile Ser Pro Gly 610 615 620 -6- 200920399

Asp His Val Met Glu 6ly Lys Lys Ala Thr Leu Ser Cys Glu Ser Asp 625 630 635 640

Ala Asn Pro Pro lie Ser Gin Tyr Thr Trp Phe Asp Ser Ser Gly Gin 645 650 655

Asp Leu His Ser Ser Gly Gin Lys Leu Arg Leu Glu Pro Leu Glu Val 660 665 670

Gin His Thr Gly Ser Tyr Arg Cys Lys Gly Thr Asn Gly lie Gly Thr 675 680 685

Gly Glu Ser Pro Pro Ser Thr Leu Thr Val Tyr Tyr Ser Pro Glu Thr 690 695 700 lie Gly Lys Arg Val Ala Leu Gly Leu Gly Phe Cys Leu Thr He Phe 705 710 715 720 lie Leu Ala lie Trp Gly Met Lys lie Gin Lys Lys Trp Lys Gin Asn 725 730 735

Arg Ser Gin Gin Gly Leu Gin Glu Asn Ser Ser Gly Gin Ser Phe Phe 740 745 750

Val Arg Asn Lys Lys Ala Arg Arg Thr Pro Leu Ser Glu Gly Pro Gin 755 760 765

Ser Gin Gly Cys Tyr Asn Pro Ala Met Asp Asp Thr Val Ser Tyr Ala 770 775 780 lie Leu Arg Phe Pro Glu Ser Asp Thr His Asn Ala Gly Asp Ala Gly 785 790 795 800

Thr Pro Ala Thr Gin Ala Pro Pro Pro Asn Asn Ser Asp Thr Val Thr 805 810 815

Tyr Ser Val Ile Gin Lys Arg Pro Met Gly Asp Tyr Glu Asn Val Asn 820 825 830

Pro Ser Cys Pro Glu Asp Glu Ser lie His Tyr Ser Glu Leu Val Gin 835 840 845

Phe Gly Ala Gly Lys Arg Pro Gin Ala Lys Glu Asp Val Asp Tyr Val 850 855 860

Thr Leu Lys His 865 &lt;210&gt; 5 &lt;211> 20 &lt;212&gt; DNA &lt; 213 &gt; 213 &gt; 220 &lt; 223 &gt; 223 &gt; Human CD22 based &lt;400&gt; 5 gcttgcacccc agacacgaca 20 200920399 &lt;210&gt 6 &lt;211> 20 &lt;212&gt; DNA &lt;213>manual&lt;220&gt;&lt;223&gt; based on human CD22 primer &lt;400&gt; 6 cctctgctgc agcccatcca 20 &lt;210> 7 &lt;211&gt; 2544 &lt;212&gt; DNA &lt;213> (new) human &lt;220&gt;&lt;221&gt; CDS &lt;222>(1).. (2544)

&lt;223> cDNA based on human CD22 &lt;400&gt; 7 atg cat etc etc Egc ecc tgg etc ctg etc ctg gtt eta gaa tac ttg 48

Met His Leu Leu Gly Pro Trp Leu Leu Leu Leu Val Leu 61u Tyr Leu 15 10 15 get ttc tet gac tea agt aaa tgg gtt ttt gag cac cct gaa acc etc 96

Ala Phe Ser Asp Ser Ser Lys Trp Val Phe GIu His Pro Glu Thr Leu 20 25 30 tac gee tgg gag ggg gee tgc gtc tgg ate ccc tgc acc tac aga gee 144

Tyr Ala Trp Glu Gly Ala Cys Val Trp lie Pro Cys Thr Tyr Arg Ala 35 40 45 eta gat ggt gac ctg gaa age ttc ate ctg ttc cac aat cct gag tat 192

Leu Asp Gly Asp Leu Glu Ser Phe lie Leu Phe His Asn Pro Glu Tyr 50 55 60 aac aag aac acc teg aag ttt gat ggg aca aga etc tat gaa age aca 240

Asn Lys Asn Thr Ser Lys Phe Asp Gly Thr Arg Leu Tyr Glu Ser Thr 65 70 75 80 aag gat ggg aag gtt cct tet gag cag aaa agg gtg caa ttc ctg gga 288

Lys Asp Gly Lys Val Pro Ser Glu Gin Lys Arg Val Gin Phe Leu Gly 85 90 95 gac aag aat aag aac tgc aca ctg agt ate cac ccg gtg cac etc aat 336

Asp Lys Asn Lys Asn Cys Thr Leu Ser lie His Pro Val His Leu Asn 100 105 110 gac agt ggt cag ctg ggg ctg agg atg gag tee aag act gag aaa tgg 384

Asp Ser Gly Gin Leu Gly Leu Arg Met Glu Ser Lys Thr Glu Lys Trp 115 120 125 atg gaa ega ata cac etc aat gtc tet gaa agg cct ttt cca cct cat 432

Met Glu Arg Ile His Leu Asn Val Ser Glu Arg Pro Phe Pro Pro His 130 135 140 ate cag etc cct cca gaa att caa gag tee cag gaa gtc act ctg acc 480 lie Gin Leu Pro Pro Glu He Gin Glu Ser Gin Glu Val Thr Leu Thr 145 150 155 160 tgc ttg ctg aat ttc tee tgc tat ggg tat ccg ate caa ttg cag tgg 528

Cys Leu Leu Asn Phe Ser Cys Tyr Gly Tyr Pro Ile Gin Leu Gin Trp 165 170 175 etc eta gag ggg gtt cca atg agg cag get get gtc acc teg acc tee 576

Leu Leu Glu Gly Val Pro Met Arg Gin Ala Ala Val Thr Ser Thr Ser 180 185 190 -8- 200920399 ttg acc ate aag tet gtc ttc acc egg age gag etc aag ttc tee cca 624

Leu Thr lie Lys Ser Val Phe Thr Arg Ser Glu Leu Lys Phe Ser Pro 195 200 205 cag tgg agt cac cat ggg aag att gtg acc tgc cag ett cag gat gca 672

Gin Trp Ser His His Gly Lys lie Val Thr Cys Gin Leu Gin Asp Ala 210 215 220 gat ggg aag ttc etc tee aat gac aeg gtg cag ctg aac gtg aag cac 720

Asp Gly Lys Phe Leu Ser Asn Asp Thr Val Gin Leu Asn Val Lys His 225 230 235 240 acc ccg aag ttg gag ate aag gtc act ccc agt gat gee ata gtg agg 768

Thr Pro Lys Leu Glu lie Lys Val Thr Pro Ser isp Ala lie VaT Arg 245 250 255 gag ggg gac tet gtg acc atg acc tgc gag gtc age age age aac ccg 816

Glu Gly Asp Ser Val Thr Met Thr Cys Glu Val Ser Ser Ser Asn Pro 260 265 270 gag tac aeg aeg gta tee tgg etc aag gat ggg acc teg ctg aag aag 864

Glu Tyr Thr Thr Val Ser Trp Leu Lys Asp Gly Thr Ser Leu Lys Lys 275 280 285 cag aat aca ttc aeg eta aac ctg ege gaa gtg acc aag gac cag agt 912

Gin Asn Thr Phe Thr Leu Asn Leu Arg Glu Val Thr Lys Asp Gin Ser 290 295 300 ggg aag tac tgc tgt cag gtc tee aat gac gtg ggc ccg gga agg teg 960

Gly Lys Tyr Cys Cys Gin Val Ser Asn Asp Val Gly Pro Gly Arg Ser 305 310 315 320 gaa gaa gtg ttc ctg caa gtg cag tat gee ccg gaa cct tee aeg gtt 1008

Giu Glu Val Phe Leu Gin Val Gin Tyr Ala Pro Glu Pro Ser Thr Val 325 330 335 cag ate etc cac tea ccg get gtg gag gga agt caa gtc gag ttt ett 1056

Gin lie Leu His Ser Pro Ala Val Glu Gly Ser Gin Val Glu Phe Leu 340 345 350 tgc atg tea ctg gee aat cct ett cca aca aat tac aeg tgg tac cac 1104

Cys Met Ser Leu Ala Asn Pro Leu Pro Thr Asn Tyr Thr Trp Tyr His 355 360 365 aat ggg aaa gaa atg cag gga agg aca gag gag aaa gtc cac ate cca 1152

Asn Gly Lys Glu Met Gin Gly Arg Thr 6!u Glu Lys Val His lie Pro 370 375 380 aag ate etc ccc tgg cac get ggg act tat tee tgt gtg gca gaa aac 1200

Lys lie Leu Pro Trp His Ala Gly Thr Tyr Ser Cys Val Ala Giu Asn 385 390 395 400 att ett ggt act gga cag agg ggc ccg gga get gag ctg gat gtc cag 1248

Ile Leu Gly Thr Gly Gin Arg Gly Pro Gly Ala Glu Leu Asp Val Gin 405 410 415 tat cct ccc aag aag gtg acc aca gtg att caa aac ccc atg ccg att 1296

Tyr Pro Pro Lys Lys Val Thr Thr Val lie Gin Asn Pro Met Pro lie 420 425 430 ega gaa gga gac aca gtg acc ett tee tgt aac tac aat tee agt aac 1344

Arg Glu Gly Asp Thr Val Thr Leu Ser Cys Asn Tyr Asn Ser Ser Asn 435 440 445 ccc agt gtt acc egg tat gaa tgg aaa ccc cat ggc gee tgg gag gag 1392

Pro Ser Val Thr Arg Tyr Glu Trp Lys Pro His Gly Ala Trp Glu Glu 450 455 460 cca teg ett ggg gtg ctg aag ate caa aac gtt ggc tgg gac aac aca 1440

Pro Ser Leu Gly Val Leu Lys lie Gin Asn Val Gly Trp Asp Asn Thr 465 470 475 480 acc ate gee tgc gca cgt tgt aat agt tgg tgc teg tgg gee tee cct 1488

Thr lie Ala Cys Ala Arg Cys Asn Ser Trp Cys Ser Trp Ala Ser Pro -9- 200920399 485 490 495 gtc gcc ctg aat gtc cag tat gcc ccc cga gac gtg agg gtc egg aaa 1536

Val Ala Leu Asn Val 6ln Tyr Ala Pro Arg Asp Val Arg Val Arg Lys 500 505 510 ate aag ccc ett tee gag att cac tet gga aac teg gtc age etc caa 1584 lie Lys Pro Leu Ser Glu lie His Ser Gly Asn Ser Val Ser Leu Gin 515 520 525 tgt gac ttc tea age age cac ccc aaa gaa gtc cag ttc ttc tgg gag 1632

Cys Asp Phe Ser Ser Ser Pro Pro Lys Glu Val Gin Phe Phe Trp Glu 530 535 540 aaa aat ggc agg ett ctg ggg aaa gaa age cag ctg aat ttt gac tee 1680

Lys Asn Gly Arg Leu Leu Gly Lys Glu Ser 6ln Leu Asn Phe Asp Ser 545 550 555 560 ate tee cca gaa gat get ggg agt tac age tgc tgg gtg aac aac tee 1728 lie Ser Pro Glu Asp Ala Gly Ser Tyr Ser Cys Trp Val Asn Asn Ser 565 570 575 ata gga cag aca geg tee aag gcc tgg aca ett gaa gtg ctg tat gca 1776

He Gly Gin Thr Ala Ser Lys Ala Trp Thr Leu Glu Val Leu Tyr Ala 580 585 590 ccc agg agg ctg cgt gtg tee atg age ccg ggg gac caa gtg atg gag 1824

Pro Arg Arg Leu Arg Val Ser Met Ser Pro Gly Asp Gin Val Met Glu 595 600 605 ggg aag agt gca acc ctg acc tgt gag age gac gcc aac cct ccc gtc 1872

Gly Lys Ser Ala Thr Leu Thr Cys Glu Ser Asp Ala Asn Pro Pro Val 610 615 620 tee cac tac acc tgg ttt gac tgg aat aac caa age etc ccc tac cac 1920

Ser His Tyr Thr Trp Phe Asp Trp Asn Asn Gin Ser Leu Pro Tyr His 625 630 635 640 age cag aag ctg aga ttg gag ccg gtg aag gtc cag cac teg ggt gcc 1968

Ser Gin Lys Leu Arg Leu Glu Pro Vai Lys Val Gin His Ser Gly Ala 645 650 655 tac tgg tgc cag ggg acc aac agt gtg ggc aag ggc cgt teg cct etc 2016

Tyr Trp Cys Gin Gly Thr Asn Ser Val Gly Lys Gly Arg Ser Pro Leu 660 665 670 age acc ett act gtc tac tat age ccg gag acc ate ggc agg cga gtg 2064

Ser Thr Leu Thr Val Tyr Tyr Ser Pro Glu Thr lie Gly Arg Arg Val 675 680 685 get gtg gga etc ggg tee tgc etc gcc ate etc ate ctg gca ate tgt 2112

Ala Val Gly Leu Gly Ser Cys Leu Ala Me Leu lie Leu Ala lie Cys 690 695 700 ggg etc aag etc cag cga cgt tgg aag agg caca age cag cag ggg 2160

Gly Leu Lys Leu Gin Arg Arg Trp Lys Arg Thr Gin Ser Gin Gin Gly 705 710 715 720 ett cag gag aat tee age ggc cag age ttc ttt gtg agg aat aaa aag 2208

Leu Gin Glu Asn Ser Ser Gly Gin Ser Phe Phe Val Arg Asn Lys Lys 725 730 735 gtt aga agg gcc ccc etc tet gaa gac ccc cac tee ctg gga tgc tac 2256

Val Arg Arg Ala Pro Leu Ser Glu Asp Pro His Ser Leu Gly Cys Tyr 740 745 750 aat cca atg atg gaa gat ggc att age tac acc acc ctg ege ttt ccc 2304

Asn Pro Met Met Glu Asp Gly lie Ser Tyr Thr Thr Leu Arg Phe Pro 755 760 765 gag atg aac ata cca cga act gga gat gca gag tee tea gag atg cag 2352

Glu Met Asn lie Pro Arg Thr Gly Asp Ala Glu Ser Ser Glu Met Gin 770 775 780 -10- 200920399 2400 2448 2496 2544 aga cct ccc egg acc tgc gat gac aeg gtc act tat tea gca ttg cac

Arg Pro Pro Arg Thr Cys Asp Asp Thr Val Thr Tyr Ser Ala Leu His 785 790 795 800 aag ege caa gtg ggc gac tat gag aac gtc att cca gat ttt cca gaa

Lys Arg Gin Val Gly Asp Tyr Glu Asn Val Ile Pro Asp Phe Pro Glu 805 810 815 gat gag ggg att cat tac tea gag ctg ate cag ttt ggg gtc ggg gag

Asp Glu Gly lie His Tyr Ser Glu Leu He Gin Phe Gly Val Gly Glu 820 825 830 egg cct cag gca caa gaa aat gtg gac tat gtg ate etc aaa cat tga

Arg Pro Gin Ala Gin Glu Asn Val Asp Tyr Val Ile Leu Lys His 835 840 845 &lt;210〉 8 &lt;211&gt; 847 &lt;212&gt; PRT &lt;213> (new) human &lt;220&gt;&lt;223&gt; CD22 amino acid sequence &lt;400&gt; 8

Met His Leu Leu Gly Pro Trp Leu Leu Leu Leu Val Leu Glu Tyr Leu 15 10 15

Ala Phe Ser Asp Ser Ser Lys Trp Val Phe Glu His Pro Glu Thr Leu 20 25 30

Tyr Ala Trp Glu Gly Ala Cys Val Trp lie Pro Cys Thr Tyr Arg Ala 35 40 45

Leu Asp Gly Asp Leu Glu Ser Phe lie Leu Phe His Asn Pro Glu Tyr 50 55 60

Asn Lys Asn Thr Ser Lys Phe Asp Gly Thr Arg Leu Tyr Glu Ser Thr 65 70 75 80

Lys Asp Gly Lys Val Pro Ser Glu Gin Lys Arg Val Gin Phe Leu Gly 85 90 95

Asp Lys Asn Lys Asn Cys Thr Leu Ser Ile His Pro Val His Leu Asn 100 105 110

Asp Ser Gly Gin Leu Gly Leu Arg Met Glu Ser Lys Thr Glu Lys Trp 115 120 125

Met Glu Arg Ile His Leu Asn Val Ser Glu Arg Pro Phe Pro Pro His 130 135 140 lie Gin Leu Pro Pro Glu He Gin Glu Ser Gin Glu Val Thr Leu Thr 145 150 155 160

Cys Leu Leu Asn Phe Ser Cys Tyr Gly Tyr Pro Me Gin Leu Gin Trp 165 170 175

Leu Leu Glu Gly Val Pro Met Arg Gin Ala Ala Val Thr Ser Thr Ser 180 185 190 -11 - 200920399

Leu Thr lie Lys Ser Val Phe Thr Arg Ser Glu Leu Lys Phe Ser Pro 195 200 205

Gin Trp Ser His His Giy Lys lie Val Thr Cys Gin Leu Gin Asp Ala 210 215 220

Asp Giy Lys Phe Leu Ser Asn Asp Thr Val Gin Leu Asn Val Lys His 225 230 235 240

Thr Pro Lys Leu Glu lie Lys Val Thr Pro Ser Asp Ala lie Val Arg 245 250 255

Glu Giy Asp Ser Val Thr Met Thr Cys Glu Val Ser Ser Ser Asn Pro 260 265 270

Glu Tyr Thr Thr Val Ser Trp Leu Lys Asp Giy Thr Ser Leu Lys Lys 275 280 285

Gin Asn Thr Phe Thr Leu Asn Leu Arg Glu Val Thr Lys Asp Gin Ser 290 295 300

Giy Lys Tyr Cys Cys Gin Val Ser Asn Asp Val Giy Pro Giy Arg Ser 305 310 315 320

Glu Glu Val Phe Leu Gin Val Gin Tyr Ala Pro Glu Pro Ser Thr Val 325 330 335

Gin lie Leu His Ser Pro Ala Val Glu 6ly Ser Gin Val Glu Phe Leu 340 345 350

Cys Met Ser Leu Ala Asn Pro Leu Pro Thr Asn Tyr Thr Trp Tyr His 355 360 365

Asn Giy Lys Glu Met Gin Giy Arg Thr Glu Glu Lys Vai His lie Pro 370 375 380

Lys lie Leu Pro Trp His Ala Giy Thr Tyr Ser Cys Val Ala Glu Asn 385 390 395 400

Me Leu Giy Thr Giy Gin Arg Giy Pro Giy Ala Glu Leu Asp Val Gin 405 410 415

Tyr Pro Pro Lys Lys Val Thr Thr Val lie Gin Asn Pro Met Pro lie 420 425 430

Arg Glu Giy Asp Thr Val Thr Leu Ser Cys Asn Tyr Asn Ser Ser Asn 435 440 445

Pro Ser Val Thr Arg Tyr Glu Trp Lys Pro His 6ly Ala Trp Glu Glu 450 455 460

Pro Ser Leu Giy Val Leu Lys Me Gin Asn Val Giy Trp Asp Asn Thr 465 470 475 480

Thr lie Ala Cys Ala Arg Cys Asn Ser Trp Cys Ser Trp Ala Ser Pro -12- 200920399 485 490 495

Val Ala Leu Asn Val Gin Tyr Ala Pro Arg Asp Val Arg Val Arg Lys 500 505 510 lie Lys Pro Leu Ser Glu lie His Ser Gly Asn Ser Val Ser Leu Gin 515 520 525

Cys Asp Phe Ser Ser Ser His Pro Lys Glu Val Gin Phe Phe Trp Glu 530 535 540

Lys Asn Gly Arg Leu Leu Gly Lys Glu Ser Gin Leu Asn Phe Asp Ser 545 550 555 560

He Ser Pro Glu Asp Ala Gly Ser Tyr Ser Cys Trp Val Asn Asn Ser 565 570 575 lie Gly Gin Thr Ala Ser Lys Ala Trp Thr Leu Glu Val Leu Tyr Ala 580 585 590

Pro Arg Arg Leu Arg Val Ser Met Ser Pro Gly Asp Gin Val Met Glu 595 600 605

Gly Lys Ser Ala Thr Leu Thr Cys Glu Ser Asp Ala Asn Pro Pro Val 610 615 620

Ser His Tyr Thr Trp Phe Asp Trp Asn Asn Gin Ser Leu Pro Tyr His 625 630 635 640

Ser Gin Lys Leu Arg Leu Glu Pro Val Lys Val Gin His Ser Gly Ala 645 650 655

Tyr Trp Cys Gin Gly Thr Asn Ser Val Gly Lys Gly Arg Ser Pro Leu 660 665 670

Ser Thr Leu Thr Val Tyr Tyr Ser Pro Glu Thr Me G!y Arg Arg Val 675 680 685

Ala Val Gly Leu Gly Ser Cys Leu Ala lie Leu lie Leu Ala lie Cys 690 695 700

Gly Leu Lys Leu Gin Arg Arg Trp Lys Arg Thr Gin Ser Gin Gin Gly 705 710 715 720

Leu Gin Glu Asn Ser Ser Gly Gin Ser Phe Phe Val Arg Asn Lys Lys 725 730 735

Val Arg Arg Ala Pro Leu Ser Glu Asp Pro His Ser Leu Gly Cys Tyr 740 745 750

Asn Pro Met Met Glu Asp Gly Me Ser Tyr Thr Thr Leu Arg Phe Pro 755 760 765

Glu Met Asn I ie Pro Arg Thr Gly Asp Ala Glu Ser Ser Glu Met Gin 770 775 780 -13- 200920399

Arg Pro Pro Arg Thr Cys Asp Asp Thr Val Thr Tyr Ser Ala Leu His 785 790 795 800

Lys Arg Gin Val Gly Asp Tyr Glu Asn Val lie Pro Asp Phe Pro Glu 805 810 815

Asp Glu Gly lie His Tyr Ser Glu Leu lie Gin Phe Gly Val Gly Glu 820 825 830

Arg Pro Gin Ala 6ln Glu Asn Val Asp Tyr Val ile Leu Lys His 835 840 845 &lt;210&gt; 9 &lt;211> 26 &lt;212&gt; DNA &lt;213>Manual&lt;220&gt;&lt;223&gt; The primer &lt;400&gt; 9 ccaagcttgc caccatgcgc gtccat 26 &lt;210&gt; 10 &lt;211> 20 &lt;212&gt; DNA &lt;213>manual&lt;220&gt;&lt;223>based on mouse CD22 primer &lt;400&gt; Atacttaaca tccagatgca 20 &lt;210> 11 &lt;211&gt; 24 &lt;212> DNA &lt;213>Artificial <220> &lt;223>Based on human CD22&lt;400> 11 accccgaagt tggagatcaa ggtc 24 &lt;210&gt; 12 &lt ;211> 20 &lt;212> DNA &lt;213>Artifical&lt;220&gt;&lt;223>Introduction based on human CD22&lt;400> 12 cttttctgct cagaaggaac 20 &lt;210&gt; 13 &lt;211> 20 &lt;212&gt; DNA &lt;213>Artifical&lt;220&gt;&lt;223&gt; Based on human CD22 primer &lt;400&gt; 13 -14- 20 200920399 cctcccaaga aggtgaccac &lt;210> 14 &lt;211> 26 &lt;212> DNA &lt;213&gt;&lt;220> <223> Based on the introduction of human CD22 &lt;400> 14 gcctgcaggt gtcaatgttt gaggat 26 &lt;210 15 &lt;211> 31 &lt;212> DNA &lt;213>manual&lt;220&gt;&lt;223>introduction based on human CD22&lt;400&gt; 15 tataccggta tgcatctcct cggcccctgg c 31 &lt;210> 16 &lt;211&gt; 34 &lt;;212>DNA&lt;213>Artificial <220> &lt;223>Introduction based on human CD22&lt;400&gt; 16 atagctagct caatgtttga ggatcacata gtcc 34 &lt;210&gt; 17 &lt;211&gt; 47 &lt;212> DNA &lt;213&gt; Artificial &lt;220> &lt;223> Based on human CD22 introduction &lt;400> 17 tatggccgga agggccttgg ccgactcaag taaatgggtt tttgagc 47 &lt;210> 18 &lt;211> 32 &lt;212> DNA &lt;213>manual&lt;220&gt;223&gt;Based on human CD22&lt;400&gt; 18 atagctagcg tgcttcacgt tcagctgcac eg 32 &lt;210> 19 &lt;211&gt; 31 &lt;212&gt; DNA &lt;213>manual&lt;220&gt;&lt;223&gt; based on mouse CD22 Primer&lt;400&gt; 19 tatgetagea tgcgcgtcca ttacctgtgg c 31 -15- 200920399 &lt;210&gt; 20 &lt;211> 53 &lt;212> DNA &lt;213> Labor &lt;220> &lt;223> Based on mouse CD22 primer &lt ;400&gt; 20 ataaagcttt caatggtgat ggtgat Gatg ccgcttgccg atggtctctg gac 53 &lt;210> 21 &lt;211> 29 &lt;212&gt; DMA &lt;213>manual <220> &lt;223> based on human CD22 introduction &lt;400> 21 tatgtatacc ccgaagttgg agatcaagg 29 &lt;210> 22 &lt;211> 34 &lt;212> DNA &lt; 213 &gt; 213 &gt; 220 &lt; 220 &lt; 223 &gt; 223> based on human CD22 &lt;400&gt; 22 tatgtatacc cctcccaaga aggtgaccac agtg 34 &lt;210&gt; 23 &lt;211&gt;;212&gt; DNA &lt;213>Artifical&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody 1B11, VH &lt;400&gt; 23 gaggtgcagc tggtggagtc tgggggaggc ctggtcaaac ctggggggtc cctgagactc 60 tcctgtgcag cctctggatt caccttcagt acctatagca tgacctgggt ccgccaggct 120 ccagggaagg gactggagtg ggtctcatcc attagtagta gtagtcgtta catatactac 180 gcagactcag tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240 ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagaaggtgg 300 ggatcgggcg cttttgatat ctggggccaa gggacaatga tcaccgtctc ttca 354 &lt; 210 &gt; 24 &lt; 211 &gt; 324 &lt; 212 &gt; DNA &lt; 213> artificial &lt; 220 &gt; &lt; 223>End Human-type anti-human CD22 antibody 1B11, VK &lt; 400 &gt; 24 gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120 cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180 -16- 200920399 gacaggttcc gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240 cctgaagatt ttgcagtgta ttactgtcag cagtatggta Gctcaccgta cacttttggc 300 caggggacca agctggagat caaa 324 &lt;210> 25 &lt;211> 351 &lt;212&gt; DNA &lt;213>manual&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody 12F6, VH &lt;400&gt; caggtgcagc tacagcagtg gggcgcagga ctgttgaagc cttcggagac cctgtccctc 60 acctgGgctg tctatggtgg gtccttcaat tttttccatt ggagctggat ccgccagocc 120 ccagggaagg ggctggagtg gattggggaa atcaatcata gtggaagcac caactacaac 180 ccgtccctca agagtcgagt caccatatca gtagacacgt ccaagaatca gttttccctg 240 aagctgagct ctgtgaccgc cgcagacacg gctgtttatt attgtgcgaa taactgggaa 300 tacaactggt tcgacccctg gggccaggga accctggtca ccgtctcctc a 351 &lt;210&gt; 26 &lt;211&gt; 321 &lt;212&gt; DNA &lt;213>manual&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody 12F6, VK &lt;400&gt; 26 gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca gggccagtca gagtgttaac agctacttag cctggtacca acagaaacct 120 ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240 gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgctcac tttcggcgga 300 gggaccaagg tggagatcaa a 321 &lt; 210> 27 &lt; 211> 351 <212> DNA &lt; 213> artificial &lt; 220 &gt; &lt; 223> fully human type anti-human CD22 antibody 13B11, VH &lt; 400> 27 gaagtgcagc tgttggagtc tgggggaggc tcggcacagc ctggggggtc cctgagactc 60 tcctgtgcag cctctggatt cgcctttagg acctatggca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagtt attagtggta gtggtggttt cacaaagtac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa Cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gaaa Gaaggg 300 ctggggattc ttgactactg gggccaggga accctggtca ccgtctcctc a 351 &lt;210> 28 -17- 200920399 &lt;211> 321 &lt;212&gt; DNA <213>&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody 13B11, VK &lt; 400 &gt; 28 gacatccaga tgacccagtc tccatcctca ctgtctgcat ctgtaggaga cagagtcacc 60 atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 120 gagaaagccc ctaagtccct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttatta ctgccaacag tataatagtt acccgtggac gttcggccaa 300 gggaccaagg tggaaatcaa a 321 &lt; 210> 29 &lt;211&gt; 354 &lt;212> DNA &lt;213>Artifical&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody17ίί5, VH &lt;400&gt; 29 caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcaggt aaatcatatg atggaagaaa taaatactat 180 gcagactccg tgaagggccg attcaccatc tccaga Galaca attccaagaa caogttgtat 240 ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gagagcctca 300 actggggact actttgacta ctggggccag ggaaccctgg tcaccgtctc ctca 354 &lt;210&gt; 30 &lt;211> 321 &lt;212&gt; DNA &lt;213>manual&lt;220&gt;&lt;223&gt; human CD22 antibodies 17H5, VK &lt; 400 &gt; 30 gaaattgtgc tgactcagtc tccagacttt cagtctgtga ctccaaagga gaaagtcacc 60 atcacctgcc gggccagtca gagcattggt agtagcttac actggtacca gcagaaacca 120 gatcagtctc caaagctcct catcaagtat gcttcccagt ccttctcagg ggtcccctcg 180 aggttcagtg gcagtggatc tgggacagat ttcacGctca ccatcaatag cctggaagct 240 gaagatgctg cagcgtatta ctgtcatcag agtagtagtt taccgtacac ttttggccag 300 gggaccaagc tggagatcaa a 321 &lt;210&gt; 31 &lt;211&gt; 354 &lt;212> DNA &lt; 213 &gt; 213 &gt; 221 &lt; 223 &lt; 223 &gt; 223 &gt; fully human type anti-human CD22 antibody 20F4, VH &lt;400&gt; 31 18- 200920399 caggtgcagc tgcaggagtc gggcccagga ctggtgaagc Cttcggagac cctgtccctc 60 acctgcactg tctctggtgc ctccatcagt agttactact ggagctggat ccggcagccc 120 ccagggaagg gactggagt g gattggatat atctattaca gtgggaacac caactacaaa 180 ccctccctct ggagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240 aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag agagaggagg 300 gactctagga tattcgatct ctggggccgt ggcaccctgg tcactgtctc ctca 354 &lt; 210> 32 &lt; 211 &gt; 321 &lt; 212 &gt; DNA &lt; 213> Artificial &lt; 220 > &lt; 223> fully human type anti-human CD22 antibody 20F4, VK &lt; 400> 32 gacatccaga tgacccagtc tccatoctca ctgtctgcat ctgtaggaga cagagtcacc 60 atcacttgtc gggcgagtca gggtgttagc agctggttag cctggtatca gcagaaacca 120 gagaaagccc ctaagtccct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttatta Ctgccaacag tataatagtt acccgtacac ttttggccag 300 gggaccaagc tggagatcaa a 321 &lt;210&gt; 33 &lt;211> 384 &lt;212> DNA &lt;213>artificial&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody 23C6, VH &lt;400 〉 33 caggtccagc tggtgcagtc tggggctgag gtgaaaaaga ctgggtcctc ggtgaaggtc 60 tcctgcaagg cttctg gagg caccttcagc agctatggta tcaactgggt gcgacaggcc 120 cctggacaag ggcttgaatg gatgggagag atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagtctac 240 atggagctga gcagcctgag agctgaggac acggccgtgt attactgtgc gagagatcag 300 ggtgtagtgg aacccactac tactactacg gtatggacgt ctggggccaa 360 gggaccacgg tcaccgtctc ctca 384 & lt tggtagctgc; 210> 34 &lt; 211> 321 &lt;212>DNA&lt;213>manual&lt;220&gt;&lt;223>Complete human type anti-human CD22 antibody 23C6, VK1 &lt;400&gt; 34 gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120 ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg Catcccagcc 180 -19- 200920399 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240 gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgtggac gttcggccaa 300 gggaccaagg tggaaatcaa a 321 &lt;210> 35 &lt;211> 321 &lt;212&gt; DNA &lt;213>manual&lt;220&gt; 223>Complete human type resistance Class-CD22 antibody 23C6, VK2 &lt; 400> 35 gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca gggccagtca gagtgttagc aacttcttag cctggtacca acagaaacct 120 ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240 gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccgac gttcggccaa 300 gggaccaagg tggaaatcaa a 321 -20-

Claims (1)

200920399 X. Patent Application 1_ An antibody 'is an antibody against a B cell-specific surface antigen and has a function of changing the production of interleukin-6 by B cells and/or has an interleukin for B cells. - ίο produces a change. The antibody according to claim 1, wherein the B cell-specific surface antigen is at least one selected from the group consisting of CD22a, CD22p, CD19, CD20, CD79a and C D 7 9 β. 3. The antibody according to claim 1 or 2, wherein the cell-specific surface antigen is CD22a and/or CD22P. 4. The antibody of claim 3, wherein the CD22a and/or CD220 is derived from a human or derived from a non-human primate. 5. The antibody according to any one of claims 1 to 4, wherein the antibody against the B cell-specific surface antigen is a monoclonal antibody. 6. The antibody according to any one of claims 1 to 5, wherein the antibody against the B cell-specific surface antigen is a mouse, a non-human primate or a human monoclonal antibody or any two of the antibodies A chimeric antibody composed of the species. 7. The antibody of claim 6, wherein the antibody to the beta cell-specific surface antigen is an isolated human type monoclonal antibody. The antibody according to any one of claims 1 to 7, which inhibits the production of interleukin-6 by B cells and/or enhances the production of interleukin-10 by B cells. The antibody according to any one of claims 1 to 8, wherein the antibody against the B cell-specific surface antigen does not have a function of reducing normal B cells. 200920399 1 0. The antibody of claim 9, wherein the normal sputum cell is reduced by 〇0/ by the antibody against the sputum cell-specific surface antigen. Above 20%. 1 1. An antibody according to any one of claims 3 to 10, wherein the antibody to CD22a and/or CD22P does not recognize the extracellular regions 1 and 2 of cD22a and/or CD220. An antibody according to claim 11 wherein an epitope for an antibody against CD22a and/or CD22p is present between c 〇 2 2 α and/or extracellular regions 3 to 7 of CD220, respectively. The antibody of claim 12, wherein an epitope for an antibody against CD22a and/or CD22p is present between cD22ct and/or an extracellular region 3 to 5 of CD22p, respectively. 14. The antibody according to claim 12, wherein an epitope for an antibody against CD22a and/or CD22P is present between the extracellular regions 5 to 7 of CD2 2a and/or C D 2 2 β, respectively. An antibody that binds to an epitope that is recognizable to a group consisting of 1B11, 12F6, 13B1 1, 17H5, 20F4, and 23C6 for CD22o [and/or CD22P] The epitope of the antibody is the same. A screening method for an antibody, which is a screening method for an antibody against a B cell-specific surface antigen, the method comprising the steps of: i) adding an antibody to a B cell-specific surface antigen to a B cell; Next, the step of changing the generation of interleukin-6 by B cells and/or the step of changing the generation of interleukin-10 by B cells is confirmed. -2 - 200920399 1 7. The screening method of claim 16 wherein the cell-specific surface antigen is selected from the group consisting of CD22a, CD22P, CD19, CD79a and CD79P. 18. The screening method according to claim 16 or 17 of the patent application, the B cell-specific surface antigen is selected from the group consisting of CD22a and/or CD22P 1 9 · as claimed in claims 16 to 18 A method in which the B cell line is selected from the group consisting of B cells, derived from rodent blood cells, derived from rodent lymphoid cells, derived from rodent splenocytes, derived from human peripheral blood cells, derived from humans Lymph node cells, human spleen cells, and groups derived from human tonsil cells. 2 如. As disclosed in any one of claims 16 to 19, B-cell interleukin-6 The production is induced and/or enhanced by B cell activity. 2 1. A screening method according to claim 20, wherein the activation of B is by at least one pre-selector selected from the group consisting of anti-IgM antibodies, anti-IgM antibody beads, Anti-IgG anti-IgG antibody beads, anti-Ig antibody, anti-Ig antibody beads, components of Staphylococcus aureus Cowan I strain, lipopolysaccharide, Pokeweed mil, human vesicular prion (Epstein-Barr virus), CpG oligodeoxynucleotide, and Toll-like receptor ligands. 2 2 · The sieving method of any one of the methods of claim 16 to 21, wherein ii) inhibits the production of interleukin-6 by B cells and/or enhances the interleukin of B cells. -10 producers. B fine CD20, wherein the end of the sputum type is from 0. The cells are in the living body, the grape cells are: 〇ge η, the oxygen nucleus is the prosthesis -3- 200920399 23 · The screening of the 22nd item of the patent scope Test method in which inhibition of B-cell interleukin-6 production only causes the disappearance of b-cell-producing b-cells and/or inhibition of interleukin-6 only on B cells capable of producing interleukin-6 produce. 24. The screening method according to any one of claims 16 to 23, wherein the change in the production of interleukin-6 is inhibited by more than 5% and less than 1% and less than 5%. 25, as claimed in claim 16 A screening method according to any one of the 23 items, wherein the change in the level of interleukin-6 is suppressed to be 25% or more and 100% or less. 2 6 . The screening method according to any one of claims 16 to 23, which confirms that the change in the production of interleukin-6 is suppressed to 4% or more and 100% or less. 2 7. A screening method according to any one of claims 16 to 26, wherein the change in the expression of interleukin-1 is enhanced by more than 50%. The method for producing an antibody is produced by subjecting an antibody obtained by the method of claims 16 to 27 or an antibody of claims 1 to 15 to a genetic recombination method. An antibody obtained by the method of any one of claims 16 to 27. 3 〇 . The antibody of claim 29, wherein the anti-system monoclonal antibody. 3 1. An antibody according to claim 29 or 30, wherein the anti-system human monoclonal antibody. -4- 200920399 3 2 _ A pharmaceutical composition comprising the antibody of any one of claims 1 to j 5 or the application of the patent scopes 29 to 31 as an active ingredient. 3 3 · If the scope of patent application is 3 2, treatment and/or prevention is selected from diseases associated with interleukin- and B lymphoid tumors. 3 4 · If you apply for a patent, you can prevent and/or treat autoimmune diseases. The pharmaceutical composition of the item, which is used for the treatment of 6 and/or interleukin-10, the stomach composition of the stomach, which is used for '^Castleman's 3 5 · as claimed Prevention and/or treatment of chronic rheumatic lupus erythematosus. 3 2 1 stomach &amp; medicinal composition, which is used to treat 4 multiple sclerosis or whole body