[go: up one dir, main page]

WO1998001580A1 - GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE - Google Patents

GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE Download PDF

Info

Publication number
WO1998001580A1
WO1998001580A1 PCT/US1996/011479 US9611479W WO9801580A1 WO 1998001580 A1 WO1998001580 A1 WO 1998001580A1 US 9611479 W US9611479 W US 9611479W WO 9801580 A1 WO9801580 A1 WO 9801580A1
Authority
WO
WIPO (PCT)
Prior art keywords
dna
protein
sequence
ser
nucleic acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1996/011479
Other languages
English (en)
Inventor
Bing Lim
Chaker N. Adra
Jean-Michel Lelias
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beth Israel Deaconess Medical Center Inc
Original Assignee
Beth Israel Deaconess Medical Center Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Beth Israel Deaconess Medical Center Inc filed Critical Beth Israel Deaconess Medical Center Inc
Priority to PCT/US1996/011479 priority Critical patent/WO1998001580A1/fr
Priority to AU64872/96A priority patent/AU6487296A/en
Publication of WO1998001580A1 publication Critical patent/WO1998001580A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • Atopic diseases which include allergy, asthma, atopic der atititis (or eczema) and allergic rhinitis, together constitute one of the largest group of clinical disorders requiring medical intervention. In the United Kingdom alone, atopy gives rise to 3-5 million cases and as many as 2,000 deaths each year.
  • Atopy is generally defined as a disorder of Immunoglobulin E (IgE) responses to common antigens, such as pollen or house dust mites. It is frequently detected by either elevated total serum IgE levels, antigen specific IgE response or positive skin tests to common allergens.
  • IgE Immunoglobulin E
  • atopy can result from dysregulation of any part of the pathway which begins with antigen exposure and IgE response to the interaction of IgE with its receptor on mast cells, the high affinity Fc receptor Fc e RI, and the subsequent cellular activation mediated by that ligand- receptor engagement (Ravetch, Nature Genetics, 7:117-118 (1994).
  • Cookson et al . Lancet, 333:1292-1295 (1989) have reported a genetic link between generalized atopic IgE responses and a locus on human chromosome llq.
  • Fc e RI is part of a tetrameric receptor complex consisting of an or chain, a ⁇ chain and two ⁇ chains (Kinet et al . , Proc. Natl . Acad. Sci . USA, 15:6483-6487 (1988)). Together, they mediate interaction with IgE-bound antigens leading to dramatic cellular responses, such as the massive degranulations of mast cells. Thought until recently to be expressed only in mast cells and basophils, the high- affinity receptor Fc € RI has been shown to be present also in Langerhans cells (Kinet, J.-P. et al . , Proc. Natl . Acad. Sci .
  • the ⁇ subunit, Fc e RI/3 is a 4-transmembrane protein with both the amino and carboxyl termini residing in the cytoplasm.
  • the human CD20 antigen Tedder, T.F., et al . , Proc. Natl . Acad. Sci . USA 55:208-212 (1988)
  • T.F. murine equivalent Ly-44
  • CD20 protein also contain four transmembrane domains with the amino and carboxyl ends on the same cytoplasmic side of the cell membrane. There is an overall amino acid similarity of 16% between CD20 and Fc.Rl ⁇ . Furthermore, the murine Fc e RI-? gene maps to the same region in chromosome 19 as the Ly-44 gene (Huppi, K. e al., J. Immunol . 143 : 3787-3791 (1989)).
  • genes that may play a role in IgE responses or atopic diseases would be desirable. It would also be desirable to develop an assay which can detect hereditary atopy.
  • the invention relates to a recombinant D ⁇ A molecule which encodes a HT m4 protein, a transfected host cell which has been stably transfected with a D ⁇ A molecule which encodes a HT rn4 protein and a recombinant HT m4 protein.
  • the invention also relates to a method for detecting the presence of a hereditary atopy.
  • Figures 1A and IB depict the cD ⁇ A sequence of the HT m4 gene and the amino acid sequence of the encoded protein.
  • ⁇ ucleotide sequence is numbered on the left.
  • the amino acid sequence of the longest open reading frame is numbered on the right beginning with the first presumed initiating methionine.
  • An upstream in-frame stop codon, TAA is indicated in bold letters at position 85.
  • a TAA stop codon (END) is followed by a 3' untranslated region containing an AATAAA poly adenylation signal.
  • the four putative transmembrane domains are underlined. Two phosphorylation sites are underlined with dotted lines.
  • the invention relates to the discovery and cloning of the HT m4 cDNA.
  • the 1672-nucleotide long cDNA contains a long open reading frame, beginning at nucleotide position 97.
  • the expression product of the cDNA is a 4 transmembrane spanning protein with a calculated molecular mass of about 25 kDa and 214 amino acids.
  • the sequences are set forth in Figures 1A and IB.
  • HT m4 contains four hydrophobic domains of 20 to 21 amino acids .
  • the amino terminal region before the beginning of the first hydrophobic domain contains four prolines.
  • Each of the hydrophilic regions between the transmembrane segments contains a single proline.
  • Several substrates for Casein kinase 2 phosphorylation (Pina, L.A. Biochim . Biophys . Acta . 1054 : 267 -284 (1990)) of serine/threonine are found at residues 24 (TGPE) , 155 (SSSE) , 181 (TLLE) , and 203 (SREE) and for Protein Kinase C phosphorylation at residue 149 (SLR) .
  • the sequence is consistent with a polypeptide chain that crosses the membrane four times, projecting two small loops extracellularly, and retaining the amino- and carboxyl- terminal portions in the cytoplasm.
  • the HT m4 protein encompasses an expression product which possesses one or more of the functions of the native protein. Included are functional protein or polypeptide fragments of the native protein and/or proteins or polypeptides where one or more amino acids have been deleted, added or substituted. Preferably, the protein or polypeptide shares at least about 50 % homology and more preferably at least about 75 % homology with the corresponding sequences of the native protein of Figures 1A and IB.
  • Recombinant DNA molecules of the invention encode an HT m4 protein, as defined herein.
  • the molecule shares at least about 50 % homology, and preferably at least about 75 % homology (such as at least about 90 % homology) with the corresponding sequences of the native gene, particularly in highly conserved regions of the 4-transmembrane protein family of HT ra4 , CD20 and Fc e RI / S.
  • the recombinant DNA molecule comprises the corresponding encoding nucleotide sequences of Figures 1A and IB .
  • recombinant DNA molecules such as probes, can be employed, for example, to isolate genes encoding transmembrane proteins or receptors, such as the Fc.RI.
  • Such molecules comprise recombinant DNA molecules which hybridize to all of or a fragment of the sequences of Figures 1A and IB.
  • the molecules hybridize under stringent conditions, such as those set forth in Sambrook et al . Molecular Clonine: A Laboratory Manual , 2nd Edition (Cold Spring Harbor Lab., Cold spring Harbor, NY (1989) .
  • the recombinant DNA molecules can contain coding and non-coding sequences.
  • the molecules comprise at least about 25 nucleotides and more preferably at least about 60 nucleotides with 95-100 % to pull out gene.
  • DNA probes comprise sequences the same as or homologous corresponding to the region encoding the N- or C-termini of the protein.
  • HT m4 The predicted structure of HT m4 as discussed herein demonstrates the relationship of this protein to Fc,RI/3 and the CD20 antigen and provides evidence for a family of 4- transmembrane spanning proteins.
  • the conservation of amino acids between all three proteins is highest in the four transmembrane domains. While much greater divergence exist in the hydrophilic amino and carboxyl termini, several amino acids within these regions are conserved such as the presence of 4 to 5 prolines in the amino terminus of all three proteins.
  • Two conserved cysteine residues in the second extracellular domain between Tm-3 and Tm-4 suggest that intra- or inter-molecular di-sulphide bonds in this domain are present in all three proteins.
  • HT m4 also contains two phosphorylation sites (threonine 24 and serine 203 ) in the cytoplasmic region of the protein. Finally, there is a well conserved SP(P) motif near the carboxyl end of all three proteins. The difference between CD20 and the other two proteins is contributed significantly by several long stretches of non-homologous amino acids.
  • the carboxyl terminus of Fc e RI/3 contains the Reth or antigen receptor activation motif (ARAM) (Reth, M. Na ture (London) 338 : 383 - 384 (1989)), which is not present in CD20 or HT m4 .
  • ARAM antigen receptor activation motif
  • the ARAM sequence is found in the cytoplasmic tail of several receptor subunits including CD3 ⁇ , ⁇ , e and ⁇ , Igo; and IgS, in MB-1 and B29 antigen, and in the ⁇ and ⁇ chain of Fc e RI (Weiss, A. and Littman, D.R. Cell 76:263-274 (1994)).
  • Tyrosine residues in ARAM sequences are believed to be critical inducers of and substrates for phosphorylation by cytoplasmic tyrosine kinases, allowing for the recruitment of additional effector molecules (Weiss, A. and Littman, D.R. Cell 76:263-274 (1994); Paolini, R. et al .
  • Chromosome mapping localized the HT m4 gene to chromosome llql2-13.1, the location of the CD20 gene.
  • the murine Fc e RI ⁇ and the murine equivalent for CD20, Ly-44 are both located in the same position in mouse chromosome 19 (Tedder, T.F. et al . , J. Immunol . 141 : 4388 - 4394 (1988); Clark, E.A. and Lane, J.L. Annu . Rev. Immunol . 9:97-127 (1991); Huppi, K. et al . , J " . Immunol . 143 : 3787- 3791 (1989)).
  • the three genes are believed to have been originated and evolved from the same locus, further supporting the proposition that they are members of the same family of related proteins. They also form a family of proteins that is quite distinct from another large family of 4-transmembrane proteins related to TAPA- KFearon, D.T. Curr. Op. Immunol . 5:341-348 (1993); Barclay, A.N. et al . , The Leucocyte Antigen Facts Book, (Academic Press Inc., San Diego, CA) (1993)) which include CD9, CD37, CD532, CD63 and R2.
  • the HT m4 gene and protein can be useful in the research and study of the induction of expression of Fc.RI and the particular function of Fc € RI / 3.
  • the HT m4 gene and protein can be useful in, for example, the design of drugs which can block or inhibit induction of Fc e RI, thereby treating atopic diseases.
  • Fc £ RI/3 was found to be associated with the low-affinity Fc receptor for IgG, Fc ⁇ RIII (CD16) , in mast cells (Kurosaki, T. et al . , J. Exp. Med . 175:447-451 (1992)).
  • Fc e RI ⁇ has also been found as a homodimer in association with Fc ⁇ RIII in macrophages (Ra, C. et al .
  • Fc £ RI ⁇ may be found as homodimers and as a heterodimer with the ⁇ - chain of T cell receptor (Letourneur, O. et al . , J. Tmmunol. 147:2652-2656 (1991)).
  • HT m4 The expression of HT m4 in all hematopoietic lineages and not in any of the non-hematopoietic cells tested indicates that HT m4 participates in biochemical pathways unique to hematopoietic lineages.
  • Nucleic acids such as DNA probes, comprising sequences of the HT m4 genes can be used in an assay to detect patients suffering from hereditary atopic disorders.
  • Either DNA or RNA can be used in the present assay method.
  • the DNA which can be used in the method can be cDNA or genomic DNA.
  • the source of DNA can be from any cell or cells removed from the individual and can include cultured progeny thereof, such as somatic cells, blood cells, sperm, fibroblasts or other somatic or germline cells. Also because the nucleic acid which is preferably analyzed is germline DNA, the method can be carried out prior or subsequent to onset of disease or disease symptoms. Where cDNA or RNA is to be used, the nucleic acid source should be from hematopoietic cells.
  • the presence of mutation can be determined using methods generally known in the art, such as by PCR (described herein below) .
  • the nucleic acid comprising the site of mutation or its complement can be sequenced, thereby identifying the presence of a mutation.
  • the protein encoded by the DNA can be sequenced or identified, thereby establishing the presence of the mutation.
  • an antibody which selectively binds to one of the mutated sequence or wild-type sequence can be made and used to screen a protein fraction for the respective proteins.
  • PCR methods using a pair of primers specific for sequence flanking the mutation region.
  • the resulting products can be sequenced, analyzed on gels, such as polyacrylamide or agarose gels, or evaluated by physical characteristics such as melting temperature or secondary structure.
  • Other methods for determining nucleic acid mutations or modifications can be employed, as well.
  • Co-amplification of two alleles in a heterozygote can generate PCR products which differ in the gene and therefore their melting and/or secondary structure characteristics are likely to differ. Under conditions as described in, e.g., Mutter and Boynton (Nucleic Acids Res. 23:1411 (1995)), amplification efficiency of the two alleles is near-equivalent, generating PCR products in a ratio proportional to that of the genomic template. Variability and biasing can be diminished by substitution of 7-deaza-2' -dGTP for dGTP during amplification, an intervention which reduces stability of intramolecular and intermolecular GC basepairing.
  • Allelic PCR fragments are easily separated, for example, by gel electrophoresis and detected by intercalating dye staining (e.g., ethidium bromide).
  • capillary electrophoresis can be employed.
  • capillary electrophoresis is in a polymer network consisting of 8% polyacryloylamino- ethoxyethanol in the absence of cross-linker, and offers a simple procedure for separation and on-line detection via UV absorbance at 254 nm, thus avoiding additional staining steps.
  • the capillary column can be used repeatedly and the electropherogram can be stored on magnetic support. Comparisons among different runs can be obtained aligning all tracings to an internal standard of a known base pair size added as a marker (Nesi et al . , Electrophoresis, 15:644-6 (1994) ) .
  • the presence of a mutation can be determined according to the method of Yamamoto et al. (Biochem. Biophys. Res. Comm. , 182:507 (1992)).
  • the DNA or RNA obtained from the subject to be tested is amplified by standard PCR, a primer extension is carried out following addition of dideoxy ATP to the reaction mixture.
  • the extension of the end-labeled reverse primer adjacent to 3' end of the site of mutation stops at a selected nucleotide after the sequence to be tested and the resultant primer products can be analyzed by denaturing polyacrylamide gel electrophoresis and autoradiography .
  • Additional PCR based methods which can be used include random rapid amplification of cDNA ends (RACE) , described by Carney et al . (Gene, 155:289, 1995); single strand confor ation polymorphism analysis (Ris-Stalpers et al . , Pediatric Res., 36:227 (1994)) and reverse transcriptase PCR (Nakamura et al . , J. Neurological Sci. 122:74 (1994)).
  • Additional hybridization techniques include the use of probes labeled with the same or different radioactive or fluorescent dyes, for example. This method allows for the direct detection of a mutation or wild type sequences (see, e.g., Sanpei et al., Biochem. Biophys. Res. Comm. 212:341-6 (1995); Taneja, J. Cell Biology, 128:995-1002 (1995) and Saito, Japanese Journal of Human Genetics, 39:421-5 (1994) ) .
  • the protein which is encoded by the gene or fragment thereof, or in the alternative, the nucleic acid can be separated by size using art-recognized separation media and methods. Standard polyacrylamide gels or a modified SDS-PAGE protocol using low concentration of methylenebisacrylamide and long runs (Ide et al . , Biochem. Biophys. Res. Comm. 209:1119 (1995)).
  • the mutation associated with hereditary atopy can be identified by isolating and sequencing the HT m 4 gene of a population of individuals suspected of having hereditary atopy and of a population of individuals believed to not possess hereditary atopy.
  • the sequences thus provided can be correlated and compared (e.g., obtaining consensus sequences or aligning sequences).
  • "Mutation”, as defined herein, is a consistent deviation in sequence shared by a population (or subpopulation thereof) of individuals suspected of having hereditary atopy, in comparison to the sequences found in the population (or subpopulation) of individuals believed to not possess hereditary atopy.
  • the nucleic acids of the invention can be useful as probes to map genes on the human chromosome, such as employing the methods of fluorescence in si tu hybridization (Kobayashi e ai., Blood, 81 : 3027-3033 (1993)).
  • the HT m4 protein can be employed in the preparation of antibodies, such as monoclonal antibodies, according to methods known in the art.
  • the antibodies can be used to block or mimic ligand binding to the receptor comprising HT m4 or other receptors, such as Fc £ RI, isolate the antibodies can be used to the HT m4 protein or hematopoietic cells which contain the HT m4 protein.
  • the antibodies can also be useful in the detection of hematopoietic cells in a sample.
  • the method comprises contacting the sample with the antibody under conditions sufficient for the antibody to bind to the HT ⁇ n4 protein and detecting the presence of bound antibody.
  • Hematopoietic cell lines used in this study included lymphomyeloid (DU528) , erythroleukemic (K562,OCIR), promyelocytic (HL60) , myeloblastic (KG-1) , monoblastic (U937) , T-cell leukemia/lymphoma (MOLT-4, Lyl7, Lyl3) and myeloma (OCI- My5) lines.
  • RNAs of neutrophils and eosinophils from normal individuals and eosinophils from a patient with hypereosinophilic syndrome were kindly provided by Dr. Peter Weller and Dr. Kaiser Lim, Harvard Medical School.
  • cDNA inserts released from two of the hematopoietic (DU528/BS-1 and KG-l/BS-1) and the non-hematopoietic (BS- 1/BS-l) subtractive libraries were purified, labeled with 32 P, and used as probes to screen the K562/BS-1 library
  • the U937 cell line was grown to a concentration of 5 x 10 s cells per ml and differentiation was induced with 50 nM phorbol 12-myristate 13-acetate (PMA; Sigma) .
  • HT ⁇ Gene Chromosomal Localization of the HT ⁇ Gene: The chromosomal location of the HT m4 gene was determined by fluorescent in situ hybridization (FISH) as previously described
  • Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) : Reverse transcriptase reaction was carried out as described (Sanger, F. et al., Proc. Natl . Acad. Sci . USA 74:5463-5467 (1977)) with R ⁇ As from cell lines or cells enriched for various cell types to obtain first strand cD ⁇ As.
  • the cD ⁇ As were subjected to PCR amplification reactions as described (Wulf, G.M. et al . , EMBO J . 12:5065-5074 (1993)), using primers spanning nucleotide 721 to 1087 of HT m4 to give a predicted PCR product of 388 nucleotides.
  • the sense primer used was 5' -TCACCTCCCAATTCTGTGTAATCAAGA-3 ' (SEQ ID NO: 1)
  • the anti-sense primer was 5' -GATTATACCGCCTTCGTTCCTTA AACC-3' (SEQ ID NO: 2) .
  • PCR reactions were carried out with 100 nM primers for 30 cycles of denaturation (1 minute at 94 °C) , annealing (1 minute at 54°C) and extension (2 minutes at 72 °C) .
  • HT m Expression pattern of HT m : A combination of Northern blot and RT-PCR analysis was used to determine the spectrum of tissue and lineage expression of the gene.
  • the cDNA insert of clone HT m4 hybridized to a transcript of about 1.7 kb in five hematopoietic lines which included myeloid and erythroid lineages and to normal human bone marrow cells.
  • the HT m4 and mRNA was not detectable in a T cell lymphoma line (Lyl7) and in a lymphomyeloid leukemic line with T and granulocytic differentiation potential (DU528) .
  • non-hematopoietic cell lines which included lung, cervical, brain, skeletal muscle, melanoma, hepatoma, and bone marrow stromal cells, no hybridizing mRNA could be detected.
  • These non-hematopoietic lines included cells of ectodermal, endodermal and mesodermal origin. The mRNA was also absent in the primary blast cells of a patient diagnosed to have M4 acute myeloid leukemia (AMD .
  • ASD M4 acute myeloid leukemia
  • RNA samples particularly those derived from cells in quantities too limited for Northern blot analysis
  • RT-PCR RNA sample-derived from cells in quantities too limited for Northern blot analysis
  • the quality of the first strand cDNAs obtained after reverse transcription was satisfactory as evaluated by using primers for the housekeeping gene HPRT.
  • oligonucleotides were synthesized and used as specific primers for PCR amplification.
  • the predicted PCR product of a 388 nucleotide-long DNA was obtained in normal bone marrow cells and the HL60 cell line but not in the HeLa and Lyl7 cell line, confirming the Northern blot analysis.
  • RNAs from a human mast cell line (HMC-1) and a megakaryocytic line (M07e) were also positive for HT m4 mRNA.
  • RNAs from normal eosinophils or neutrophils and eosinophils from a patient with hype -eosinophilie syndrome (HES) also yielded the predicted PCR product.
  • Two leukemic T-cell lines (Lyl3 and MOLT4) and a myeloma cell line (OCI-My5) were also found to be positive for HT m4 . All PCR-derived DNAs hybridized positively to radio-labeled HT m4 in subsequent Southern analysis.
  • HT ml human cDNA The nucleotide sequence of the HT m4 cDNA and the predicted amino acid sequence are shown in Figures 1A and IB.
  • the 1672- nucleotide long cDNA contains a long open reading frame, beginning at nucleotide position 97, encoding a protein of 214 amino acids with a calculated molecular mass of 25 kDa.
  • a hydrophilicity analysis with the Kyte-Doolittle algorithm (Kyte, J. and Doolittle, R.F. J. Mol . Biol . 157:105-132 (1982)), reveals that HT m4 contains four hydrophobic domains of 20 to 21 amino acids.
  • the amino terminal region before the beginning of the first hydrophobic domain contains four prolines. Each of the hydrophilic regions between the transmembrane segments contains a single proline.
  • Several substrates for Casein kinase 2 phosphorylation (Pina, L.a. Biochim. Biophys . Acta 1054 : 267- 284 (1990)) of serine/threonine are found at residues 24 (TGPE) , 155 (SSSE) , 181 (TLLE) , and 203 (SREE) and for Protein Kinase C phosphorylation at residue 149 (SLR) .
  • the sequence is consistent with a polypeptide chain that crosses the membrane four times, projecting two small loops extracellularly, and retaining the amino- and carboxyl-terminal portions in the cytoplasm.
  • HT ml during differentiation of cell line U937 To ascertain if expression of the HT m4 mRNA may vary depending on the stage of cellular differentiation, we examined the consequences of induced differentiation in the monoblastic cell line U937. Exposure of the cells to PMA rapidly induced differentiation to macrophages, as confirmed morphologically and molecularly by monitoring the marker for terminally differentiated macrophages, CDllb (Arnout, M.A. Immunol . Rev. 114:145-180 (1990)). The expression of HT m4 mRNA over a period of 48 hours showed an initial increase followed by a down regulation so that by day three, HT m4 transcripts were detectable at a very low level .
  • HT rnl is located on chromosome llql2-13: Forty-one chromosomes from 30 metaphases were scored for the positive chromosomal band. Band llql2 was labeled on eighteen of chromosome 11 homologues, band llql3.1 on twenty-one of chromosome 11 homologues and band llql3.2 on two of chromosome 11 homologues. No signal was detected on other chromosomes in these cells. Similar results were obtained in an additional experiment using this probe. Thus, HT m4 is localized to chromosome Ilql2-ql3.1.
  • MOLECULE TYPE DNA (genomic)
  • MOLECULE TYPE DNA (genomic)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention concerne une molécule ADN recombinante encodant pour une protéine HTm4, une cellule hôte transformée ayant été transfectée de manière stable avec une molécule d'ADN codant pour une protéine HTm4 et une protéine HTm4 recombinante. L'invention a également trait à une méthode de détection de la présence d'une éventuelle atopie à caractère héréditaire.
PCT/US1996/011479 1996-07-10 1996-07-10 GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE Ceased WO1998001580A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/US1996/011479 WO1998001580A1 (fr) 1996-07-10 1996-07-10 GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE
AU64872/96A AU6487296A (en) 1996-07-10 1996-07-10 Recombinant htm4 gene, protein and use thereof in methods for detecting hereditary atopy

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US1996/011479 WO1998001580A1 (fr) 1996-07-10 1996-07-10 GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE

Publications (1)

Publication Number Publication Date
WO1998001580A1 true WO1998001580A1 (fr) 1998-01-15

Family

ID=22255433

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/011479 Ceased WO1998001580A1 (fr) 1996-07-10 1996-07-10 GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE

Country Status (2)

Country Link
AU (1) AU6487296A (fr)
WO (1) WO1998001580A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115029383A (zh) * 2022-04-21 2022-09-09 苏天生命科技(苏州)有限公司 Ms4a3蛋白在调控红细胞成熟中的应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995005481A1 (fr) * 1993-08-18 1995-02-23 Isis Innovation Limited Procede de diagnostic et therapie
US5552312A (en) * 1994-10-06 1996-09-03 Beth Israel Hospital Boston Recombinant HTm4 gene, protein and assays

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995005481A1 (fr) * 1993-08-18 1995-02-23 Isis Innovation Limited Procede de diagnostic et therapie
US5552312A (en) * 1994-10-06 1996-09-03 Beth Israel Hospital Boston Recombinant HTm4 gene, protein and assays

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ADRA C ET AL: "Cloning of the cDNA for a hematopoeic cell-specific protein related to CD20 and the beta subunit of the high affinity IgE receptor: Evidence for a family of proteins with four membrane-spanning regions", PROC. NATL. ACAD. SCI. USA, vol. 91, October 1994 (1994-10-01), pages 10178 - 82, XP002026149 *
ROY A ET AL: "Molecular scanning of human diseases", SINGAPORE JOURNAL OF OBSTETRICS AND GYNAECOLOGY, vol. 26, no. 3, November 1993 (1993-11-01), pages 176 - 86, XP000600337 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115029383A (zh) * 2022-04-21 2022-09-09 苏天生命科技(苏州)有限公司 Ms4a3蛋白在调控红细胞成熟中的应用
CN115029383B (zh) * 2022-04-21 2024-02-20 苏天生命科技(苏州)有限公司 Ms4a3蛋白在调控红细胞成熟中的应用

Also Published As

Publication number Publication date
AU6487296A (en) 1998-02-02

Similar Documents

Publication Publication Date Title
Itokawa et al. A structural polymorphism of human dopamine D2 receptor D2 (Ser311→ Cys)
Hara et al. Structure and evolution of four POU domain genes expressed in mouse brain.
Deichmann et al. Common polymorphisms in the coding part of the IL4-receptor gene
JP4463329B2 (ja) Ｃａｄａｓｉｌに関与する遺伝子、診断方法および治療への適用
DeSilva et al. The human reelin gene: isolation, sequencing, and mapping on chromosome 7.
Rouillac et al. Transcript analysis of D category phenotypes predicts hybrid Rh D-CE-D proteins associated with alteration of D epitopes
AU724681B2 (en) Human genes
AU693944B2 (en) Allelic variation of the serotonin 5HT2C receptor
Adra et al. Cloning of the cDNA for a hematopoietic cell-specific protein related to CD20 and the beta subunit of the high-affinity IgE receptor: evidence for a family of proteins with four membrane-spanning regions.
WO1998058958A2 (fr) Polypeptide jagged humain, acides nucleiques codants et procedes d'utilisation
Walker et al. A novel multi-gene family of sheep gamma delta T cells
Wan et al. Conserved Chromosomal Location and Genomic Structure of Human and Mouse Fatty-Acid Amide Hydrolase Genes and Evaluation ofclasperas a Candidate Neurological Mutation
US5972688A (en) HTm4 methods of treatment and assays, agonists and antagonists
Iwasaki et al. Identification of microsatellite markers near the human genes encoding the β-cell ATP-sensitive K+ channel and linkage studies with NIDDM in Japanese
WO1995005481A1 (fr) Procede de diagnostic et therapie
US5552312A (en) Recombinant HTm4 gene, protein and assays
Coon et al. Search for mutations in the β1 GABAA receptor subunit gene in patients with schizophrenia
WO1998001580A1 (fr) GENE RECOMBINANT HTm4, PROTEINE ET UTILISATION DANS DES PROCEDES DE DETECTION D'ATOPIE HEREDITAIRE
US20030171566A1 (en) Basal cell carcinoma tumor suppressor gene
Gassner et al. Fy^ x is associated with two missense point mutations in its gene and can be detected by PCR-SSP
Vollmer et al. Molecular cloning of the critical region for glomerulopathy with fibronectin deposits (GFND) and evaluation of candidate genes
AU745234B2 (en) Genetic test for equine severe combined immunodeficiency disease
HUP9903661A2 (hu) Eljárás a humán G-fehérje béta3-alegységének génjében található génváltozás alkalmazására betegségek diagnosztizálásában
US5518880A (en) Methods for diagnosis of XSCID and kits thereof
WO1994020641A1 (fr) Methodes de diagnostic et de traitement de l'alymphocytose

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BB BG BR BY CA CH CN CZ DE DK EE ES FI GB GE HU IL IS JP KE KG KP KR KZ LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TR TT UA UG UZ VN AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): KE LS MW SD SZ UG AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

NENP Non-entry into the national phase

Ref country code: JP

Ref document number: 1998505171

Format of ref document f/p: F

NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase