JP2001502359A - Sialoadhesin family 4 (SAF-4) cDNA - Google Patents

Abstract

  (57) [Summary] Disclosed are SAF-4 polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques. Also disclosed are methods of using SAF-4 polypeptides and polynucleotides for therapy, and diagnostic assays for such uses.

Description

Description: FIELD OF THE INVENTION The present invention relates to newly identified polypeptides, polynucleotides encoding the polypeptides, treatment of the polypeptides and polynucleotides. The present invention relates to the use in identifying compounds that are agonists, antagonists and / or inhibitors that may be effective in the treatment or for therapy, and methods for producing the polypeptides and polynucleotides. BACKGROUND OF THE INVENTION The drug discovery process is currently undergoing a radical change. Because it extends to "functional genomics", high-throughput (high-efficiency) genomic or gene-based biology. This approach is rapidly replacing a relatively early approach based on “positional cloning”. The phenotype, ie, biological function or genetic disease, will be identified and the etiological gene will then be located based on its genetic map location. Functional genetic science relies heavily on a variety of bioinformatics tools to identify gene sequences of interest from many currently available molecular biology databases. There still exists a need to identify and characterize yet-unknown genes and their related polypeptides / proteins as targets for drug discovery. SUMMARY OF THE INVENTION The present invention relates to SAF-4, particularly SAF-4 polypeptides and polynucleotides, recombinant materials, and methods for producing the same. In another embodiment, the present invention relates to cancer, inflammation, autoimmune disease, allergy, asthma, rheumatoid arthritis, central nervous system inflammation, cerebellar degeneration, Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic side Chordosclerosis, head injury damage, and other neurological abnormalities, septic shock, sepsis, stroke, osteoporosis, osteoarthritis, ischemic reperfusion injury, cardiovascular disease, kidney disease, liver disease, ischemic Injury, myocardial infarction, hypotension, hypertension, AIDS, myelodysplastic syndrome and other hematological abnormalities, aplastic anemia, male baldness, and bacterial, fungal, protozoal and viral infections The present invention relates to methods for using the polypeptides and polynucleotides, including the treatment of diseases (hereinafter collectively referred to as "the diseases"). In another aspect, the invention provides a method of identifying agonists and antagonists / inhibitors using the agents provided by the invention, and treating a condition associated with SAF-4 imbalance using the identified compounds. About. In yet another embodiment, the present invention relates to diagnostic assays for detecting diseases associated with inappropriate SAF-4 activity or SAF-4 levels. DESCRIPTION OF THE INVENTION In one embodiment, the present invention relates to SAF-4 polypeptides. This kind of peptide has at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity, even more preferably to the amino acid sequence of SEQ ID NO: 2 over the entire length of SEQ ID NO: 2. Comprises an isolated polypeptide comprising an amino acid sequence having at least 95% identity, most preferably at least 97-99% identity. Such polypeptides include polypeptides comprising the amino acid sequence of SEQ ID NO: 2. Other peptides of the invention have at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity to the amino acid sequence of SEQ ID NO: 2 over the entire length of SEQ ID NO: 2. Or more preferably at least 95% identity, most preferably at least 97-99% identity. Such polypeptides include polypeptides consisting of the amino acid sequence of SEQ ID NO: 2. Further peptides of the invention include an isolated polypeptide encoded by a polynucleotide comprising the nucleotide sequence contained in SEQ ID NO: 1. The polypeptides of the present invention are considered to be members of the sialoadhesin family. Therefore, they are interesting. This is because sialoadhesin, a protein of the sialoadhesin family, CD33, CD22 and myelin-binding glycoprotein (MAG) are used as cell-interacting molecules. These proteins specifically bind to carbohydrates on target cells in a sialic acid-dependent manner. Its extracellular domain consists of several immunoglobulin-like domains of V-like and C2-like subtypes, and the homology of the intracellular portion to any signaling motif is not known. Sialoadhesin expression is restricted to macrophages, has 17 Ig-like domains, and the specific recognition sequence on target cells is Neu5Aca2,3 Galb13GalNAc. Known target cells are developing myeloid cells in the bone marrow and lymphocytes in the spleen and lymph nodes (Crocker, PR, et al., EMBO J 1994, 13: 4490-4503). CD22 is expressed only on B cells and has the a and b isoforms (with 5 and 7 Ig-like domains, respectively). CD22 is known to bind T cells, B cells, monocytes, granulocytes and erythrocytes by recognizing Neu5 Aca2,6Galbl, 4Glc (NAc) in N-linked glycans (Crocker, PR. Et al.). Stamenkovic, I. and Seed, B. Nature, 1990, 345: 74-77; Wilson, GL. Et al., J Exp Med, 1991, 173: 137-146). Since myelin binding glycoprotein (MAG) is expressed by Schwann cells in the peripheral nervous system and oligodendrocytes in the central nervous system, MAG is thought to be involved in cell adhesion to axons. MAG has two alternatively spliced variants, a large MAG (L-MAG) and a small MAG (S-MAG), each of which during embryonic development or It is expressed in any of the adults. This alternative splicing results in expression of the same extracellular domain but different intracellular domains (Pedraza, L. et al., JCB, 1990, 111: 2651-2661). CD33 is most related to SAF-4. Because they are the most closely related of all members of the family. Normally, CD33 is expressed on differentiating myeloid monocytic lines. Although CD33 is not present on early stem cells, granulocyte, erythroid, monocyte, and megakaryocyte colony forming units (CFU-GEMM) and granulocyte and mononuclear phagocyte progenitor cell colony forming units (CFU-GEMM). GM). CD33 is down-regulated by mature granulocytes but retained by mature monocytes and macrophages (Andrews, RG. Et al., Blood, 1983, 62: 124; Griffin, JD. Et al., Leuk Res 1984, 8: 521). Because CD33 has two Ig-like domains, it more readily binds targets expressing NeuAca2,3GAl in N-linked and O-linked glycans. CD33 maps to chromosome 19q13.1-13.3 in the genome that is closely associated with MAG and CD22 (Freeman, SD. Et al., Blood, 1995,85: 2005-2012). In addition, CD33 has been found to be expressed on leukemic myeloblasts in about 85% of those who have acute myeloid leukemia (AML), and AML and acute lymphoblastic leukemia (AML) ALL) is often used to distinguish it. In the treatment of AML, monoclonal antibodies against CD33 have been used therapeutically ex vivo to clear residual myeloblasts from autologous bone marrow grafts (Robertson, MJ et al., Blood, 1992 79: 2229- 2236). More recently, humanized monoclonal antibodies to CD33 have been evaluated in the in vivo treatment of AML (Caron, PC. Et al., Blood, 1994, 83: 1760-1768). These properties are hereinafter referred to as “SAF-4 activity” or “SAF-4 polypeptide activity” or “biological activity of SAF-4”. These activities include the antigenic activity and the immunological activity of the SAF-4 polypeptide, particularly the antigenic activity and the immunological activity of the polypeptide of SEQ ID NO: 2. Preferably, the polypeptide of the present invention exhibits at least one biological activity of SAF-4. The polypeptides of the present invention may be in the form of the "mature" protein or may be part of a larger protein, such as a fusion protein. It is often advantageous to include additional amino acid sequences, such as secretory or leader sequences, prosequences, sequences useful for purification such as multiple histidine residues, or stable during recombinant production. There are additional arrangements to ensure the performance. Also included in the invention are variants of the above polypeptides, ie, polypeptides that differ from a reference polypeptide by conservative amino acid substitutions (where one residue is replaced with another having similar properties). . Typical such substitutions are between Ala, Val, Le u and Ile; between Ser and Thr; between the acidic residues Asp and Glu; between Asn and Gln; between the basic residues Lys and Arg; or Occurs between the aromatic residues Phe and Tyr. In particular, a mutant in which several, 5 to 10, 1 to 5, 1 to 3, 1 to 2 or 1 amino acids are substituted, deleted or added in any combination is preferable. The polypeptide of the present invention can be produced by any appropriate method. Such polypeptides include isolated natural polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. It is. The means for producing such polypeptides are well understood in the art. In a further aspect, the present invention relates to SAF-4 polynucleotides. Such a polynucleotide has at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity to the amino acid sequence of SEQ ID NO: 2 over the entire length of SEQ ID NO: 2, Preferably, an isolated polynucleotide comprising a nucleotide sequence encoding a polypeptide having at least 95% identity is included. In this regard, polypeptides having at least 97% identity are more preferred, while those with at least 98-99% identity are even more preferred, and those with at least 99% identity are most preferred. . Such polynucleotides include polynucleotides comprising the nucleotide sequence contained in SEQ ID NO: 1 encoding the polypeptide of SEQ ID NO: 2. Further polynucleotides of the invention include at least 70% identity, preferably at least 80% identity, more preferably at least 80% identity over the entire coding region to the nucleotide sequence encoding the polypeptide of SEQ ID NO: 2. An isolated polynucleotide comprising a nucleotide sequence having at least 90% identity, more preferably at least 95% identity, is included. In this regard, polynucleotides having at least 97% identity are more preferred, while those with at least 98-99% identity are even more preferred, and those with at least 99% identity are most preferred. . Additional polynucleotides of the present invention include at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity to the polynucleotide of SEQ ID NO: 1 over the entire length of SEQ ID NO: 1. And more preferably an isolated polynucleotide comprising a nucleotide sequence having at least 95% identity. In this regard, polynucleotides having at least 97% identity are more preferred, while those with at least 98-99% identity are even more preferred, and those with at least 99% identity are most preferred. . Such polynucleotides include a polynucleotide comprising the polynucleotide of SEQ ID NO: 1 and a polynucleotide of SEQ ID NO: 1. The invention also provides polynucleotides that are complementary to all of the above polynucleotides. The nucleotide sequence of SEQ ID NO: 1 shows homology to CD33 (Simmons, D. and Seed, B., JI 141: 279 7-2800, 1988). The nucleotide sequence of SEQ ID NO: 1 is a cDNA sequence and includes a polypeptide coding sequence (nucleotide numbers 51 to 1970) encoding a polypeptide consisting of 639 amino acids, which is a polypeptide of SEQ ID NO: 2. Even though the nucleotide sequence encoding the polypeptide of SEQ ID NO: 2 is identical to the polypeptide coding sequence contained in SEQ ID NO: 1, the polypeptide of SEQ ID NO: 2 is still duplicated due to the redundancy (degeneracy) of the genetic code. It may be a sequence other than the sequence contained in SEQ ID NO: 1 that encodes. The polypeptide of SEQ ID NO: 2 is structurally related to other proteins in the sialoadhesin family and has homology to CD33 (Simmons, D. and Seed, B., JI 141: 2797-2800, 1988). And / or have structural similarity. Preferred polypeptides and polynucleotides of the present invention are expected to have, among other things, similar biological functions / properties as the polypeptides and polynucleotides homologous thereto. Further, preferred polypeptides and polynucleotides of the present invention have at least one SAF-4 activity. The present invention also relates to partial or other polynucleotides and polypeptides that were initially identified prior to the determination of the corresponding full length sequences of SEQ ID NO: 1 and SEQ ID NO: 2. Thus, in a further aspect, the invention provides (a) at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity to the nucleotide sequence of SEQ ID NO: 3 over the entire length of SEQ ID NO: 3 % Identity, more preferably at least 95% identity, most preferably 97-99% identity; (b) at least 70 nucleotides of SEQ ID NO: 3 over the entire length of SEQ ID NO: 3; A nucleotide sequence having at least 80% identity, preferably at least 80% identity, more preferably at least 90% identity, even more preferably at least 95% identity, and most preferably 97-99% identity; c) the polynucleotide of SEQ ID NO: 3, or (d) the amino acid sequence of SEQ ID NO: 4 over the entire length of SEQ ID NO: 4 With at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity, even more preferably at least 95% identity, and most preferably 97-99% identity. A nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 3, and a polynucleotide of SEQ ID NO: 3. Further, the invention provides (a) at least 70% identity, preferably at least 80% identity, more preferably at least 90% identity to the amino acid sequence of SEQ ID NO: 4 over the entire length of SEQ ID NO: 4; More preferably, a polypeptide comprising an amino acid sequence having at least 95% identity, most preferably 97-99% identity; (b) at least with respect to the amino acid sequence of SEQ ID NO: 4 over the entire length of SEQ ID NO: 4 An amino acid sequence having 70% identity, preferably at least 80% identity, more preferably at least 90% identity, even more preferably at least 95% identity, and most preferably 97-99% identity. (C) a polypeptide comprising the amino acid of SEQ ID NO: 4, and (d) a polypeptide of SEQ ID NO: 4. That polypeptides, as well as polypeptides encoded by a polynucleotide comprising the nucleotide sequence contained in SEQ ID NO: 3, to provide. The nucleotide sequence of SEQ ID NO: 3 and the peptide sequence encoded thereby are derived from an Expressed Sequence Tag (EST) sequence. One skilled in the art will appreciate that there will necessarily be some nucleotide sequence reading errors in the EST sequence (see Adams, MD et al., Nature 377 (supp) 3, 1995). Accordingly, the nucleotide sequence of SEQ ID NO: 3 and the peptide sequence encoded thereby are subject to the same inherent limitations in sequence accuracy. In addition, the peptide sequence encoded by SEQ ID NO: 3 may contain regions identical to the protein with the highest homology or structural similarity, or regions of high homology and / or structural similarity (eg, conservative amino acid differences). Contains. The polynucleotides of the present invention can be obtained by standard cloning and screening techniques from cDNA libraries derived from mRNA in cells of human primary dendritic cells by EST analysis (Adams, MD. Et al., Science ( 1991) 252: 1 651-1656; Adams, MD et al., Nature (1992) 355: 632-634; Adams, MD et al., Nature (1995) 377 Supp: 3-174). In addition, the polynucleotide of the present invention can be obtained from a natural source such as a genomic DNA library, and can also be synthesized using a commercially available known technique. When a polynucleotide of the present invention is used for recombinant production of a polypeptide of the present invention, the polynucleotide may include the coding sequence of the mature polypeptide alone or other coding sequences (eg, a leader or secretory sequence, a pre- -Encoding the pro- or pre-pro-protein sequence, or other fusion peptide portion), in the same reading frame as the mature polypeptide. For example, a marker sequence that facilitates purification of the fusion polypeptide can be encoded. In a preferred embodiment of this aspect of the invention, the marker sequence is provided by the pQE vector (Qiagen, Inc.) and Gentz et al., Proc. Natl. Acad. Sci. Hexa-histidine peptide, or HA tag, as described in USA (1 989) 86: 821-824. The polynucleotide may also include 5 'and 3' non-coding sequences, for example, transcribed but not translated sequences, splicing and polyadenylation signals, ribosome binding sites, and mRNA stabilizing sequences. As further specific examples of the present invention, several, for example, 5 to 10, 1 to 5, 1 to 3, 1 to 2, or 1 amino acid residues may be substituted or deleted in any combination. Alternatively, there is an added polynucleotide encoding a polypeptide variant comprising the amino acid sequence of SEQ ID NO: 2. A polynucleotide identical or sufficiently identical to the nucleotide sequence contained in SEQ ID NO: 1 is used to isolate full length cDNA and genomic clones encoding a polypeptide of the present invention, and To isolate cDNA and genomic clones of other genes with high sequence similarity (including homologues from non-human species and genes encoding orthologs), Or as a primer for a nucleic acid amplification (PCR) reaction. Generally, these nucleotide sequences are 70%, preferably 80%, more preferably 90%, and most preferably 95% identical to the reference nucleotide sequence. Probes or primers usually contain 15 or more nucleotides, preferably 30 or more, and may have 50 or more nucleotides. Particularly preferred probes are those having a range of 30 to 50 nucleotides. Polynucleotides encoding the polypeptides of the present invention (including homologues and orthologs derived from non-human species) can be prepared using a labeled probe having the sequence of SEQ ID NO: 1 or a fragment thereof under stringent hybridization conditions. The method comprises the steps of: screening an appropriate library below and isolating a full-length cDNA and a genomic clone containing the polynucleotide sequence. Such hybridization techniques are known to those skilled in the art. Preferred stringent hybridization conditions are 50% formamide, 5 × SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5 × Denhardt solution, 10% dextran sulfate and 20 μg / ml. Incubated overnight at 42 ° C. in a solution containing denatured and sheared salmon sperm DNA, and then washing the filters at about 65 ° C. in 0.1 × SSC. Thus, the present invention also includes a polynucleotide obtained by screening an appropriate library under stringent hybridization conditions using a labeled probe having the sequence of SEQ ID NO: 1 or a fragment thereof. As will be appreciated by those skilled in the art, in many cases, the isolated cDNA sequence will be incomplete, since the region encoding the polypeptide is truncated at the 5 'end of the cDNA. It is due to the reverse transcriptase, which originally has low "processivity" (a measure of the enzyme's ability to remain bound to the template during the polymerization reaction), and the mRNA template during first-strand cDNA synthesis. DNA copy cannot be completed. There are several methods known and available to those skilled in the art for obtaining full-length cDNAs or elongating short cDNAs, such as those based on the rapid cDNA end amplification method (RACE) (eg, Frohman et al., PNAS USA 85, 8998-9002, 1988). Recent improvements in the above techniques, as demonstrated, for example, by Marathon ^™ technology (Clontech Laboratories Inc.), have greatly simplified the search for longer cDNAs. In the Marathon ^™ technology, cDNA is made from mRNA extracted from a given tissue, and an “adapter” sequence is ligated to each end. Subsequently, nucleic acid amplification (PCR) is performed using a combination of gene-specific and adapter-specific oligonucleotide primers to amplify the "deleted" 5 'end of the cDNA. Next, a “nested” primer, a primer designed to anneal inside the amplification product (typically an adapter-specific primer that anneals further 3 ′ to the adapter sequence and an additional 5 ′ to the known gene sequence) The PCR reaction is repeated using a gene-specific primer that anneals to the side. The product of this reaction is then analyzed by DNA sequencing and the product is directly linked to the existing cDNA to complete the sequence, or another full-length sequence using new sequence information for 5 'primer design. By performing PCR, a full-length cDNA can be constructed. The recombinant polypeptides of the present invention can be produced from genetically engineered host cells containing the expression system using methods known in the art. Thus, in a further aspect, the present invention relates to an expression system containing one or more polynucleotides of the present invention, a host cell engineered with said expression system, and the production of a polypeptide of the present invention by recombinant methods. Cell-free translation systems can also be used to produce such proteins using RNA derived from the DNA constructs of the present invention. For recombinant production, the host cells are genetically engineered to incorporate the polynucleotide expression system of the invention, or a portion thereof. Introduction of polynucleotides into host cells is described in Davis et al., Basic Methods in Molecular Biology (1986) and Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY ( 1989) can be performed by methods described in many standard laboratory manuals. Suitable such methods include, for example, calcium phosphate transfection, DEAE-dextran mediated transfection, transvection, microinjection, cationic lipid mediated transfection, electroporation, transduction, scrape loading, bullet loading. There is a ballistic introduction or infection. Representative examples of suitable hosts include bacterial cells (eg, Streptococcus, Staphylococcus, E. coli, Streptomyces, Bacillus subtilis), fungal cells (eg, yeast, Aspergillus), insect cells (eg, Drosophila S2, Spodoptera Sf9). ), Animal cells (eg, CHO, COS, HeLa, C127, 3T3, BHK, HEK293, Bowes melanoma cells) and plant cells. A wide variety of expression systems can be used. Such expression systems include, for example, systems derived from chromosomes, episomes and viruses, such as those derived from bacterial plasmids, bacteriophages, transposons, yeast episomes, insertion factors, yeast chromosomal elements, viruses (eg, baculovirus, SV40 Such as papovavirus, vaccinia virus, adenovirus, fowlpox virus, pseudorabies virus, and retrovirus), and vectors derived from combinations thereof, for example, plasmids such as cosmids and phagemids and bacteriophage genetics. Some come from elements. These expression systems may contain control regions that regulate as well as cause expression. Generally, any system or vector that can maintain, augment, and express a polynucleotide for production of the polypeptide in the host can be used. The appropriate nucleotide sequence can be inserted into the expression system by any of the commonly used and known techniques, as described in Sambrook et al., Molecular Cloning: A Laboratory Manual (supra). Appropriate secretion signals can be incorporated into the polypeptide of interest to secrete the translated protein into the endoplasmic reticulum lumen, into the periplasmic space, or into the extracellular environment. These signals may be endogenous to the polypeptide of interest or heterologous signals. When it is desired to express a polypeptide of the invention for use in a screening assay, it is generally preferred that the polypeptide be produced on the surface of cells. If so, the cells are harvested prior to use in the screening assay. If the polypeptide is secreted into the medium, the medium is recovered to recover and purify the polypeptide. If produced intracellularly, it is necessary to first lyse the cell and then recover the polypeptide. To recover and purify the polypeptide of the present invention from recombinant cell culture, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, Known methods including hydroxylapatite chromatography and lectin chromatography can be used. Most preferably, high performance liquid chromatography is used for purification. When the polypeptide is denatured during intracellular synthesis, isolation and / or purification, it is possible to restore the active conformation using known techniques for regenerating the protein. The present invention also relates to the use of the polynucleotide of the present invention as a diagnostic. Detection of a variant of a gene characterized by a polynucleotide of SEQ ID NO: 1 associated with a dysfunction is useful for diagnosing a disease caused by under-expression, over-expression or altered expression of the gene or susceptibility to the disease. It will provide a diagnostic tool that can be added or diagnosed. Individuals with mutations in the gene can be found at the DNA level by various techniques. Diagnostic nucleic acids can be obtained from cells of a subject, such as cells from blood, urine, saliva, tissue biopsy or autopsy material. Genomic DNA may be used directly for detection, or may be enzymatically amplified using PCR or other amplification methods prior to analysis. RNA or cDNA can be used in a similar manner. Deletions and insertions can be detected by a change in the size of the amplified product when compared to the normal genotype. Point mutations can be identified by hybridizing the amplified DNA to a labeled SAF-4 nucleotide sequence. Perfectly matched sequences and mismatched duplexes can be distinguished by RNase digestion or by differences in melting temperatures. DNA sequence differences can also be detected by changes in the electrophoretic mobility of DNA fragments on gels with or without denaturing agents, or by direct DNA sequencing (eg, Myers et al., Science (1985)). 230: 1242). Sequence changes at specific positions can also be confirmed by nuclease protection assays (eg, RNase and S1 protection) or by chemical cleavage (Cotton et al., Proc. Natl. Acad. Sci. USA (1985) 85: 4397-4401). checking). In another embodiment, an array of oligonucleotide probes comprising a SAF-4 nucleotide sequence or a fragment thereof can be constructed, for example, for efficient screening of genetic mutations. Array techniques are known and have general applicability and have been used to solve various molecular genetics problems, including gene expression, genetic linkage and genetic variability (eg, M See Chee et al., Science, Vol. 274, pp. 610-613 (1996)). The diagnostic assay provides a method for diagnosing or determining susceptibility to the disease by detecting a mutation in the SAF-4 gene by the method described above. In addition, the disease can be diagnosed by a method of measuring an abnormal decrease or increase in the level of polypeptide or mRNA from a sample obtained from a subject. The decrease or increase in expression can be determined at the RNA level by any of the polynucleotide quantification methods known in the art, such as nucleic acid amplification (eg, PCR, RT-PCR), RNase protection, Northern blotting, and other hybridization methods. Can be measured. Assays that can be used to determine levels of a protein, such as a polypeptide of the present invention, in a sample obtained from a host are well-known to those of skill in the art. Such assays include radioimmunoassays, competitive binding assays, western blot analysis, ELISA assays and flow cytometry analysis. Thus, in another embodiment, the invention provides: (a) a polynucleotide of the invention (preferably the nucleotide sequence of SEQ ID NO: 1) or a fragment thereof; (b) a nucleotide sequence complementary to the nucleotide sequence of (a) (C) an antibody against the polypeptide of the present invention (preferably, the polypeptide of SEQ ID NO: 2) or a fragment thereof, or (d) an antibody against the polypeptide of the present invention (preferably, the polypeptide of SEQ ID NO: 2). A diagnostic kit. It will be appreciated that in such a kit, (a), (b), (c) or (d) is a substantial component. Such kits may be susceptible to illness or illness, especially cancer, inflammation, autoimmune allergy, allergy, asthma, rheumatoid arthritis, central nervous system inflammation, cerebellar degeneration, Alzheimer's disease, Parkinson's disease, multiple sclerosis Amyotrophic lateral sclerosis, head injury damage, and other neurological abnormalities, septic shock, sepsis, stroke, osteoporosis, osteoarthritis, ischemic reperfusion injury, cardiovascular disease, kidney disease, Liver disease, ischemic injury, myocardial infarction, hypotension, hypertension, AIDS, myelodysplastic syndrome and other hematological abnormalities, aplastic anemia, male pattern baldness, and bacterial, fungal, protozoan It is useful for diagnosing infectious diseases and viral infections, or susceptibility to these. The nucleotide sequence of the present invention is also useful for chromosome identification. This sequence can specifically target a particular location on an individual human chromosome and hybridize to that particular location. Creating a chromosomal map of related sequences according to the present invention is an important first step in correlating these sequences with gene-related diseases. Once a sequence has been mapped to a precise chromosomal location, the physical location of that sequence on that chromosome can be correlated with genetic map data. This kind of data is, for example, McKusick, found in Mendelian Inheritance in Man (available online at Johns Hopkins University Welch Medical Library). Thereafter, the relationship between the gene mapped to the same chromosomal region and the disease is confirmed by linkage analysis (co-inheritance of physically adjacent genes). Differences in the cDNA or genomic sequence between affected and unaffected individuals can also be determined. If a mutation is observed in some or all of the affected individuals but not in any normal individuals, then the mutation may be responsible for the disease. The polypeptide of the present invention, a fragment or analog thereof, or a cell expressing the same can also be used as an immunogen for producing an antibody immunospecific for the polypeptide of the present invention. By "immunospecific" is meant that the antibody exhibits a substantially higher affinity for the polypeptides of the present invention than its affinity for other related polypeptides in the prior art. An antibody against the polypeptide of the present invention can be obtained by administering a fragment, analog or cell containing the polypeptide or epitope to an animal (preferably a non-human animal) using a conventional protocol. For preparation of monoclonal antibodies, any technique which provides antibodies produced by continuous cell line cultures can be used. Examples include the hybridoma method (Kohler, G. and Milstein, C., Nature (1975) 256: 495-497), the trioma method, the human B cell hybridoma method (Kozbor et al., Immunology Today (1983) 4: 72) and the EBV-hybridoma method (Cole et al., Monoclonal Antibodies and Cancer Therapy, pp. 77-96, Alan R. Liss, Inc., 1985). To make single-chain antibodies to a polypeptide of the invention, methods for preparing single-chain antibodies as described in US Pat. No. 4,946,778 can be adapted. Also, transgenic mice or other organisms, including other mammals, can be used to express humanized antibodies. Using the antibody described above, a clone expressing the polypeptide can be isolated and identified, or the polypeptide can be purified by affinity chromatography. Antibodies to the polypeptides of the present invention may have potential use, inter alia, in the treatment of said diseases. In addition, antibodies to SAF-4 polypeptides, to sub-characterize the cell population that is differentiating into the hematopoietic system, and to express SAF-4 as a diagnostic marker to distinguish different forms of cancer. As a tool to promote the ex vivo expansion (proliferation and / or differentiation) of hematopoietic progenitor cells expressing SAF-4 to clear cells from bone marrow ex vivo, It can be used as a stimulus in vivo and as a chemoprotectant in vivo. In a further aspect of the invention, the invention relates to a polypeptide or a fragment thereof according to the invention and various parts of the constant regions of the heavy or light chains of immunoglobulins of the various subclasses (IgG, IgM, IgA, IgE). And a genetically engineered soluble fusion protein comprising: The immunoglobulin is preferably the constant part of the heavy chain of human IgG, especially IgG1, in which case the fusion takes place in the hinge region. In certain instances, the Fc portion can be easily removed by incorporating a cleavage sequence that can be cleaved by blood coagulation factor Xa. Furthermore, the invention relates to methods for the genetic engineering of these fusion proteins and their use in drug screening, diagnosis and therapy. In another approach, a soluble form of the SAF-4 polypeptide that is still capable of binding the ligand in competition with endogenous SAF-4 may be administered. A typical embodiment of such a competitor comprises a fragment of a SAF-4 polypeptide. In one embodiment, the extracellular domain of SAF-4 fused to the human immunoglobulin Fc region is used, so that this domain can be used to treat cancer, inflammation, autoimmune diseases, allergies, etc. . In addition, SAF-4 / Fc polypeptides can be used as tools to promote the ex vivo expansion (proliferation and / or differentiation) of hematopoietic progenitor cells expressing SAF-4 ligand, for the recruitment of stem cells to the periphery. It can be used as an in vivo stimulus and as an in vivo chemoprotectant to wipe out SAF-4 ligand expressing cancer cells from bone marrow ex vivo. A further aspect of the present invention relates to a polynucleotide encoding such a fusion protein. Examples of fusion protein technology can be found in International Patent Applications WO94 / 29458 and WO94 / 22914. A further aspect of the present invention relates to a method of eliciting an immunological response in a mammal, the method comprising the step of eliciting an antibody and / or a T-cell immune response sufficient to protect the animal from the disease, in particular. Inoculating a mammal with a polypeptide of the formula (I). Yet another aspect of the invention directs the expression of a polynucleotide encoding a polypeptide of the invention in vivo to elicit an immunological response that produces an antibody that protects the mammal from the disease. A method of eliciting an immunological response in a mammal, comprising providing the polypeptide via a vector. A further aspect of the present invention relates to an immunological / vaccine formulation (composition) which, when introduced into a mammalian host, elicits an immunological response against the polypeptide of the present invention in the mammal, wherein the composition comprises Polypeptide or polynucleotide. The vaccine formulation may further comprise a suitable carrier. Since the polypeptide may be broken down in the stomach, it is preferably administered parenterally (eg, by subcutaneous, intramuscular, intravenous or intradermal injection). Formulations suitable for parenteral administration include sterile aqueous and non-aqueous injections, which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the recipient, and suspensions. There are aqueous and non-aqueous sterile suspensions which may contain agents or thickeners. Such formulations may be presented in single or multi-dose containers (eg, sealed ampules and vials) and may also be stored in a lyophilized condition which requires only the addition of a sterile liquid carrier immediately before use. it can. The vaccine formulation may include an adjuvant system to enhance the immunogenicity of the formulation, such as an oil-in-water adjuvant system or other adjuvant systems known in the art. The dose varies with the specific activity of the vaccine, but can be easily determined by routine experimentation. The polypeptides of the present invention have been implicated in a variety of biological functions, including a number of pathological conditions, particularly those diseases. Therefore, it is desirable to develop screening methods that identify compounds that stimulate or suppress the function of this polypeptide. Accordingly, in a further aspect, the present invention provides a method of screening a compound to identify a compound that stimulates or suppresses the function of the polypeptide. Generally, agonists or antagonists are used for the purpose of treating and preventing the above-mentioned diseases. Compounds can be identified from a variety of sources, such as mixtures of cells, cell-free preparations, chemical libraries and natural products. The agonist, antagonist or inhibitor so identified may optionally be a natural or modified substrate, ligand, receptor, enzyme, etc., of the polypeptide, and its structural or functional mimetic. (See Coligan et al., Current Protocols in Immunology 1 (2): Chapter 5 (1991)). In the screening method, the binding of the candidate compound to the polypeptide of the present invention, or a cell or membrane carrying the polypeptide, or a fusion protein thereof is easily performed using a label directly or indirectly bound to the candidate compound. Can be measured. Alternatively, the screening method may use competition with a labeled competitor. Furthermore, such screening methods can test whether a candidate compound results in a signal resulting from activation or suppression of the polypeptide, using a detection system suitable for cells carrying the polypeptide. . Generally, inhibitors of activation are assayed in the presence of a known agonist to determine the effect of the presence of the candidate compound on activation by the agonist. Constitutively active polypeptides are used in screening methods for inverse agonists or inhibitors by examining whether a candidate compound inhibits activation of a polypeptide in the absence of an agonist or inhibitor. Furthermore, in these screening methods, a candidate compound and a solution containing the polypeptide of the present invention are mixed to form a mixture, the SAF-4 activity in the mixture is measured, and the SAF-4 activity of the mixture is determined as a standard. Simply include each step of comparing with. In high throughput screening assays to identify antagonists of the polypeptides of the present invention, fusion proteins such as those made from the Fc portion and SAF-4 polypeptides described above can also be used (D. Bennett et al., J. Am. Mol. Recognition, 8: 52-58 (1995) and K. Johanson et al., J. Biol. Chem., 270 (16): 9459-9471 (1995)). In addition, a screening method for detecting the effect of an additional compound on the production of mRNA or polypeptide in cells can be assembled using the polynucleotide, polypeptide or antibody against the polypeptide of the present invention. For example, an ELISA assay for measuring the level of polypeptide secretion or cell binding using monoclonal or polyclonal antibodies according to standard methods known in the art can be constructed, comprising appropriately engineered cells. Alternatively, it can be used for searching for a substance that suppresses or enhances the production of a polypeptide from a tissue (also referred to as an antagonist or an agonist, respectively). If membrane bound or soluble receptors are present, the polypeptides of the invention can be used to identify such receptors by standard receptor binding methods known in the art. it can. Such receptor binding methods include, but are not limited to, ligand binding assays and cross-linking assays, in which the polypeptide is labeled with a radioactive isotope (eg, ¹²⁵ I) or chemically modified (eg, ¹²⁵ I). Eg, biotinylated) or fused to a peptide sequence suitable for detection and purification and incubated with a putative receptor source (cells, cell membranes, cell supernatants, tissue extracts, body fluids, etc.). Other methods include biophysical methods such as surface plasmon resonance and spectroscopy. These screening methods can also be used to identify agonists or antagonists of the polypeptide that compete for binding to the polypeptide or its receptor, if present. Standard methods for performing screening assays are well understood in the art. Examples of potential antagonists of the polypeptides of the present invention include antibodies, and in some cases, oligonucleotides or proteins (eg, ligands, ligands, etc.) that are closely related to the ligands, substrates, receptors, enzymes, etc., of the polypeptide. Fragments of a substrate, receptor, enzyme, etc.) or small molecules that bind to a polypeptide of the invention but do not elicit a response (and thus prevent the activity of the polypeptide). Thus, in another aspect, the invention is directed to identifying agonists, antagonists, ligands, receptors, substrates, enzymes, etc., of the polypeptides of the invention, or compounds that decrease or increase the production of such polypeptides. Regarding the screening kit, the kit comprises (a) the polypeptide of the present invention, (b) a recombinant cell expressing the polypeptide of the present invention, (c) a cell membrane expressing the polypeptide of the present invention, or (d) an antibody against the polypeptide of the present invention, wherein said polypeptide is preferably the polypeptide of SEQ ID NO: 2. It will be appreciated that in such a kit, (a), (b), (c) or (d) is a substantial component. Those skilled in the art will readily appreciate that the polypeptides of the present invention can also be used in methods for designing agonists, antagonists or inhibitors of the polypeptides based on their structure. This method comprises the steps of (a) first analyzing the three-dimensional structure of the polypeptide, (b) assuming the three-dimensional structure of a likely reactive or binding site of an agonist, antagonist or inhibitor; Synthesizing a candidate compound that is expected to bind or react with the selected reactive or binding site, and (d) determining whether the candidate compound is in fact an agonist, antagonist or inhibitor. It will be further appreciated that this is usually an interactive process. In a further embodiment, the present invention relates to any of an excess or a deficiency of SAF-4 polypeptide activity, such as cancer, inflammation, autoimmune disease, allergy, asthma, rheumatoid arthritis, central nervous system inflammation. Cerebellar degeneration, Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, head injury damage, and other neurological abnormalities, septic shock, sepsis, stroke, osteoporosis, osteoarthritis Ischemic reperfusion injury, cardiovascular disease, kidney disease, liver disease, ischemic injury, myocardial infarction, hypotension, hypertension, AIDS, myelodysplastic syndrome and other hematological abnormalities, aplastic anemia, male pattern Provided are treatments for baldness and abnormal conditions such as bacterial, fungal, protozoal and viral infections. If the activity of the polypeptide is in excess, several approaches are available. One approach is to suppress the function of the polypeptide, for example, by blocking the binding of ligands, substrates, receptors, enzymes, etc., or by reducing an abnormal condition by suppressing a second signal. Administering an inhibitor compound (antagonist) as described above to a patient in an amount effective to do so with a pharmaceutically acceptable carrier. In another approach, soluble forms of the polypeptide that are still capable of binding ligands, substrates, enzymes, receptors, etc., in competition with the endogenous polypeptide can be administered. A typical example of such a competitor is a fragment of a SAF-4 polypeptide. In yet another approach, expression of the gene encoding endogenous SAF-4 polypeptide can be suppressed using expression blocking techniques. These known techniques require the use of antisense sequences produced in the body or administered separately (eg, Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression (oligodeoxynucleotides as antisense inhibitors of gene expression), CRC O'Connor, J. Neurochem (1991) 56: 560 in Press, Boca Raton, FL (1988)). Alternatively, oligonucleotides that form a triple helix with this gene can be provided (eg, Lee et al., Nucleic Acids Res (1979) 6: 3073; Cooney et al., Science (1988) 241: 456; Dervan et al., Science). (1991) 251: 1360). These oligomers can be administered as such, or the related oligomers can be expressed in vivo. For treating abnormal conditions related to an under-expression of SAF-4 and its activity, several approaches are also available. One approach involves administering to a patient a therapeutically effective amount of a compound that activates a polypeptide of the invention (ie, the agonist) together with a pharmaceutically acceptable carrier to alleviate the abnormal condition. It becomes. Alternatively, gene therapy can be used to produce SAF-4 endogenously in the relevant cells of the patient. For example, a polynucleotide of the invention may be engineered for expression by a replication defective retroviral vector as described above. The retroviral expression construct is then isolated and introduced into packaging cells transduced with a retroviral plasmid vector containing RNA encoding a polypeptide of the present invention. As a result, the packaging cells will produce infectious viral particles containing the gene of interest. These producer cells are administered to a patient for in vivo cell manipulation and in vivo polypeptide expression. For an overview of gene therapy, see Chapter 20, Gene Therapy and other Molecular Genetic-based Therapeutic Approaches in Human Molecular Genetics, T Strachan and AP Read, BIOS Scientific Publishers Ltd (1996) (and references therein). That. Another approach is to administer a therapeutic amount of a polypeptide of the invention together with a suitable pharmaceutical carrier. In a further aspect, the invention provides a therapeutically effective amount of a polypeptide (eg, a soluble form of the polypeptide of the invention), an agonist or antagonist peptide, or a small molecule compound in a pharmaceutically acceptable carrier or excipient. And a pharmaceutical composition containing the same. Such carriers include, but are not limited to, saline, saline, dextrose, water, glycerol, ethanol, and combinations thereof. The invention further relates to pharmaceutical packs and kits comprising one or more containers filled with one or more components of the composition of the invention as described above. The polypeptides and other compounds of the present invention may be used alone or in conjunction with other compounds, eg, therapeutic compounds. The pharmaceutical composition can be adapted to the route of administration, for example by a systemic or an oral route. A suitable form for systemic administration is infusion (injection), typically intravenous. Other injection routes, such as subcutaneous, intramuscular or intraperitoneal, can also be used. Alternative means of systemic administration include transmucosal and transdermal administration using penetrants such as bile salts, fusidic acid, and other surfactants. In addition, oral administration is possible provided the polypeptide or other compound of the present invention can be formulated as a cerebral solvent or capsule. These compounds may be administered topically and / or localized in the form of an ointment, paste, gel or the like. The required dosage range depends on the choice of the peptide or other compound of the invention, the route of administration, the nature of the formulation, the condition of the patient and the judgment of the physician. However, suitable dosages are in the range of 0.1-100 μg / kg of patient weight. Given the variety of compounds available and the differing efficiencies of the routes of administration, it is expected that the required dosage will vary widely. For example, oral administration would be expected to require higher dosages than intravenous administration. Such variation in dosage level can be adjusted using standard empirical optimization procedures, as is well understood in the art. The polypeptide used for the treatment can also be produced in the body of a patient in the above-mentioned treatment called “gene therapy”. For example, cells from a patient are genetically engineered ex vivo with a polynucleotide such as DNA or RNA encoding the polypeptide, for example, using a retroviral plasmid vector. Thereafter, these cells are introduced into the patient. Polynucleotide and polypeptide sequences provide a valuable source of information in identifying alternative sequences having similar homology. This is facilitated by storing such sequences in a computer readable medium and then using the stored data to search a sequence database with known search tools such as GCG. Accordingly, in a further aspect, the present invention provides a computer readable medium having stored thereon a polynucleotide comprising the sequence of SEQ ID NO: 1 and / or a polypeptide encoded thereby. The following definitions are intended to facilitate understanding of terms that are often used in the above description. As used herein, “antibody” includes polyclonal and monoclonal antibodies, chimeric antibodies, single-chain antibodies, humanized antibodies, as well as Fab fragments, including Fab or other products of an immunoglobulin expression library. "Isolated" means altered "by the hand of man" from the natural state. If an "isolated" composition or substance occurs in nature, it has been changed or separated from its original environment, or both. For example, a polynucleotide or polypeptide naturally occurring in the body of a living animal is not "isolated", but a polynucleotide or polypeptide separated from its naturally occurring co-localized material is described herein. As used herein, "isolated". "Polynucleotide" generally refers to any polyribonucleotide or polydeoxyribonucleotide, which can be unmodified RNA or DNA, or modified RNA or DNA. “Polynucleotide” includes, but is not limited to, single- and double-stranded DNA, DNA in which single- and double-stranded regions are mixed, single- and double-stranded RNA, single-stranded RNA with mixed region and double-stranded region, hybrid molecule containing DNA and RNA (may be single-stranded or more typically double-stranded, where single-stranded and double-stranded regions are mixed May be included). In addition, "polynucleotide" refers to triple-stranded regions consisting of RNA or DNA or both RNA and DNA. The term "polynucleotide" also includes DNAs or RNAs containing one or more modified bases, and DNAs or RNAs with backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and special bases such as inosine. Various modifications can be made to DNA and RNA. Thus, "polynucleotide" embraces chemically, enzymatically or metabolically modified forms of polynucleotides which are commonly found in nature, as well as DNA and RNA chemical forms characteristic of viruses and cells. . "Polynucleotide" also embraces relatively short polynucleotides, often referred to as oligonucleotides. "Polypeptide" refers to a peptide or protein comprising two or more amino acids linked by a peptide bond or a modified peptide bond (ie, a peptide isostere). "Polypeptide" refers to both short chains (commonly referred to as peptides, oligopeptides or oligomers) and long chains (commonly referred to as proteins). Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. "Polypeptides" include amino acid sequences that have been modified by natural processes, such as post-translational processing, or by any of the chemical modification methods known in the art. Such modifications are elaborated in basic textbooks, more detailed scholarly articles and research literature. Modifications can be made anywhere in the polypeptide, including the peptide backbone, amino acid side chains, amino or carboxyl termini. The same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides can be branched for ubiquitination or cyclic, with or without branching. Cyclic, branched or branched cyclic polypeptides can result from post-translation natural processes and can also be produced by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, flavin covalent bond, heme moiety covalent bond, nucleotide or nucleotide derivative covalent bond, lipid or lipid derivative covalent bond, phosphatidylinositol covalent bond. , Cross-linking, cyclization, disulfide bond formation, demethylation, covalent cross-link formation, cystine formation, pyroglutamate formation, formylation, γ-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination Transfer RNA-mediated addition of amino acids to proteins such as, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulphation, arginylation, ubiquitination, etc. , Proteins-Structure and Molecular Properties 2nd E d., TE. Creighton, WH. Freeman and Company, New York, 1993; Posttranslational Covalent Modification of Proteins, BC, edited by Johnson, Academic Press, New York, 1998, Wold, F., Post-translational Protein. Modifications: Perspectives and Prospects, pgs. 1-12; Seifter et al., “Analysis for protein modifications and nonprotein cofactors”, Meth Enzymol (1990) 182: 626-646; and Rattan et al., “Protein Synthesis: Post-translational Modifications and Aging ", Ann NY Acad Sci (1992) 663: 48-62). As used herein, “variant” refers to a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide, but retains essential properties. A typical variant of a polynucleotide differs in nucleotide sequence from a reference polynucleotide. Changes in the nucleotide sequence of this variant may or may not alter the amino acid sequence of the polypeptide encoded by the reference polynucleotide. Nucleotide changes can result in amino acid substitutions, deletions, additions, fusions and truncations of the polypeptide encoded by the reference sequence, as described below. A typical variant of a polypeptide differs in amino acid sequence from a reference polypeptide. In general, the sequences of the reference polypeptide and the variant are generally closely similar and limited to differences so that they will be identical in many regions. A variant and reference polypeptide may differ in amino acid sequence by one or more substitutions, deletions, or additions in any combination. The amino acid residues to be substituted or added may or may not be those encoded by the genetic code. The variant of the polynucleotide or polypeptide may be a naturally occurring variant, such as an allelic variant, or a variant that is not known to occur naturally. Non-naturally occurring variants of polynucleotides and polypeptides can be made by mutagenesis methods or by direct synthesis. "Identity" as known in the art, is the relationship between two or more polypeptide or polynucleotide sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as determined by the match between strands of such sequences. "Identity" can be calculated without difficulty by known methods. Examples of such methods include Computational Molecular Biology, Les, AM, Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Pro Jects, Smith, DW. Ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, AM. and Griffin, HG. Ed., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, v on Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J.S. Ed., M Stockton Press, New York, 1991: and Carillo, H .; and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988), but not limited thereto. Methods for determining identity are designed to give the largest match between the sequences considered. In addition, methods for determining identity have been compiled into publicly available computer programs. Computer program methods for determining identity between two sequences include the GCG program package (Devereux, J. et al., Nucleic Acids Research 12 (1): 387 (1984)), BLAST P, BLASTN, and FASTA (Atschul, SF et al.). , J. Molec. Biol. 215: 4043-410 (1990)). The BLAST X program is generally available from NC BI and other sources (BLAST Manual, Altschul, S. et al., NCBI NLM NIH Bethesda, MD 20894; Altschul, S. et al., J. Mol. Biol. 215: 403). -410 (1990)). The known Smith Waterman algorithm can also be used to determine identity. Parameters for comparing polypeptide sequences include: l) Algorithm: Needleman & Wunsch, J .; Mol. Biol. 48: 443-453 (1970) Comparative matrix: BLOSSUM62, Hentikoff and Hentikoff, Proc. Natl. Acad. Sci. USA, 89: 10915-10919 (1992) Gap penalty: 12 Gap length penalty: 4 A program useful with these parameters is generally available as the "gap" program from the Genetics Computer Group (Madison WI). The above parameters are default parameters for peptide comparison (no terminal gap penalty). Parameters for comparing polynucleotide sequences include the following: 1) Algorithm: Needleman & Wunsch, J .; Mol. Biol. 48: 443-453 (1970); Comparison matrix: match = + 10, mismatch = 0 Gap penalty: 50 Gap length penalty: 3 A useful program with these parameters is available as the "gap" program from the Genetics Computer Group (Madlson WI). It is. These are the default parameters for nucleic acid comparison. Suitable implications for the "identity" of polynucleotides and polypeptides are optionally provided in (1) and (2) below. (1) Specific examples of the polynucleotide comprise a polynucleotide sequence having at least 50, 60, 70, 80, 85, 90, 95, 97 or 100% identity to the reference sequence of SEQ ID NO: 1. It further includes an isolated polynucleotide. The polynucleotide sequence may be the same as the reference sequence of SEQ ID NO: 1, or may include up to an integer number of nucleotide variations with respect to the reference sequence. The mutation is selected from the group consisting of deletions, substitutions (including transitions and transversions) or insertions of at least one nucleotide, such mutations being at the 5 ′ or 3 ′ terminal position of the reference nucleotide sequence, or at these terminal positions. Between the nucleotides in the reference sequence, individually or as one or more contiguous groups within the reference sequence. The number of nucleotide variations is determined by multiplying the total number of nucleotides of SEQ ID NO: 1 by the respective percent identity value (divided by 100) and subtracting the product from the total number of nucleotides of SEQ ID NO: 1, ie, _n n ≦ x n - it can be obtained by (x _n · y). Wherein, n _n is the number of nucleotide alterations, x _n is the total number of nucleotides in SEQ ID NO: 1, y is about 0.70,80% for 0.60,70% for 0.60 for 60% for 50% for 0.90,95% for 0.85,90% for 0.80,85% 1.00 for 0.97 or 100% for 0.95,97%, • is the symbol for the multiplication operator, the x _n and y Non-integer products are rounded down to the nearest integer before subtracting the product from _xn . The alteration of the polynucleotide sequence encoding the polypeptide of SEQ ID NO: 2 results in a nonsense, missense or frameshift mutation in the coding sequence, thereby altering the polypeptide encoded by the polynucleotide after such mutation. be able to. By way of example, a polynucleotide sequence of the present invention is identical to the reference sequence of SEQ ID NO: 2, ie, has 100% identity to the reference sequence or has a percent identity of less than 100%. It can include up to an integer number of amino acid mutations relative to the reference sequence. The mutation is selected from the group consisting of a deletion, substitution (including transition and transversion) or insertion of at least one nucleic acid, wherein the mutation is at the 5 'or 3' end position of the reference polynucleotide sequence, or at these ends. It can be anywhere between positions and can be intervening individually between nucleic acids in the reference sequence, or as one or more contiguous groups within the reference sequence. The number of nucleic acid mutations is determined by multiplying the total number of amino acids of SEQ ID NO: 2 by the respective percent identity (divided by 100) and subtracting the product from the total number of amino acids of SEQ ID NO: 2, ie: _n n ≦ x n - it can be obtained by (x _n · y). Wherein, n _n is the number of amino acid alterations, x _n is the total number of amino acids in SEQ ID NO: 2, y is about 70% for example and the like 0.85 for 0.80,85% for 0.70,80% ,. Are symbols representing the multiplicative operator, and the non-integer product of _xn and y is rounded down to the nearest integer before subtracting the product from _xn . (2) Specific examples of polypeptides include polypeptides having at least 50, 60, 70, 80, 85, 90, 95, 97 or 100% identity to the reference sequence of SEQ ID NO: 2. Further comprising an isolated polypeptide. The polypeptide sequence may be the same as the reference sequence of SEQ ID NO: 2, or may include up to an integer number of amino acid mutations with respect to the reference sequence. Such mutations are selected from the group consisting of deletions, substitutions (including conservative and non-conservative amino acid substitutions) or insertions of at least one amino acid, wherein the mutations are at the amino or carboxy terminal position of the reference polypeptide sequence. Or anywhere between these terminal positions, and may be interspersed individually between amino acids in the reference sequence, or as one or more contiguous groups within the reference sequence. The number of amino acid mutations is determined by multiplying the total number of amino acids in SEQ ID NO: 2 by the respective percent identity (divided by 100) and subtracting the product from the total number of amino acids in SEQ ID NO: 2, Required by _{n a ≦ x a - (x} a · y) wherein, n _a is the number of amino acid alterations, x _a is the total number of amino acids in SEQ ID NO: 2, y is about 0.60 for 60% for 50% Is 0.60, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95%, 0.97 for 97% or 1.00 for 100%, Is the symbol for the multiplicative operator, where the product of non-integers of x _a and y is rounded down to the nearest integer before subtracting the product from x _a . By way of example, a polypeptide sequence of the present invention is identical to the reference sequence of SEQ ID NO: 2, ie, has 100% identity to the reference sequence, or has a% identity less than 100%. The sequence can include up to an integer number of amino acid variations. Such mutations are selected from the group consisting of deletions, substitutions (including conservative and non-conservative amino acid substitutions) or insertions of at least one amino acid, wherein the mutations are at the amino or carboxy terminal position of the reference polypeptide sequence. Or anywhere between these terminal positions, and may be interspersed individually between amino acids in the reference sequence, or as one or more contiguous groups within the reference sequence. The number of amino acid mutations is determined by multiplying the total number of amino acids in SEQ ID NO: 2 by the respective percent identity (divided by 100), and subtracting the product from the total number of amino acids in SEQ ID NO: 2, Required by n _a ≦ x _a - in (x _a · y) formula, n _a is the number of amino acid alterations, x _a is the total number of amino acids in SEQ ID NO: 2, y 0.70,80% is about 70% for example Is 0.80 for 85%, 0.85 for 85%, etc. is a symbol representing a multiplicative operator, and the product of non-integers of x _a and y is converted to the closest integer before subtracting the product from x _a Round down. "Fusion protein" refers to a protein encoded by two, often unrelated, fused genes or fragments thereof. As an example, EP-A-0 464 describes a fusion protein comprising various parts of the constant region of an immunoglobulin molecule and another human protein or a part thereof. In many cases, for use in therapy and diagnosis, it will be advantageous to use the immunoglobulin Fc region as part of a fusion protein, which will improve pharmacokinetic properties, for example, EP-A- 0232 262). On the other hand, for some uses, it may be desirable to remove the Fc portion after expressing, detecting and purifying the fusion protein. All publications, including patents and patent application specifications, cited herein are hereby incorporated by reference in their entirety, as if each publication was clearly and individually indicated. Shall be.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 25/00 A61P 29/00 29/00 35/00 35/00 C07K 14/705 C07K 14/705 16 / 18 16/18 16/28 16/28 C12N 1/15 C12N 1/15 1/19 1/19 1/21 1/21 C12P 21/02 C 5/10 G01N 33/15 Z 15/09 ZNA 33/50 Z C12P 21/02 33/53 D G01N 33/15 C12N 15/00 ZNAA 33/50 5/00 A 33/53 A61K 37/02 (81) Designated countries EP (AT, BE, CH, CY, DE, DK , ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE) , SN, TD, TG), AP (GH, GM, KE, LS, M , SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY , CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE, GH, GM, GW, HU, ID, IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK , SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZW (72) Inventor Ericsson-Miller, Connie, Lynn United States 19341 Exton, Pennsylvania, Brineard Place 608

Claims (1)

[Claims] 1. An isolated polypeptide selected from the group consisting of the following (i) to (iii):   (i) at least the amino acid sequence of SEQ ID NO: 2 over the entire length of SEQ ID NO: 2       (a) 70% identity,       (b) 80% identity,       (c) 90% identity, or       (d) 95% identity,   An isolated polypeptide comprising an amino acid sequence having (ii) an isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or (iii) an isolated polypeptide consisting of the amino acid sequence of SEQ ID NO: 2. 2. An isolated polynucleotide selected from the group consisting of the following (i) to (vi):   Or a nucleotide sequence complementary to the isolated polynucleotide:   (i) at least the amino acid sequence of SEQ ID NO: 2 over the entire length of SEQ ID NO: 2       (a) 70% identity,       (b) 80% identity,       (c) 90% identity, or       (d) 95% identity,     Comprising a nucleotide sequence encoding a polypeptide having     A polynucleotide, (ii) a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2;     Over the entire length, at least       (a) 70% identity,       (b) 80% identity,       (c) 90% identity, or       (d) 95% identity,     An isolated polynucleotide comprising a nucleotide sequence having (iii) less than the nucleotide sequence of SEQ ID NO: 1 over the entire length of SEQ ID NO: 1     With       (a) 70% identity,       (b) 80% identity,       (c) 90% identity, or       (d) 95% identity,     An isolated polynucleotide comprising a nucleotide sequence having (iv) a simple sequence comprising a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2.     An isolated polynucleotide, (vi) an isolated polynucleotide consisting of the polynucleotide sequence of SEQ ID NO: 1     ,and (vi) stringent hybridization conditions     Below, screened with a labeled probe having the sequence of SEQ ID NO: 1 or a fragment thereof.     Isolated polynucleotide obtained by performing the above-described steps. 3. An antibody immunospecific for the polypeptide of claim 1. 4. A method of treating a subject, comprising:   (i) the subject needs to increase the activity or expression of the polypeptide of claim 1;     If       (a) administering to the subject a therapeutically effective amount of an agonist for the polypeptide         Administering, and / or       (b) an isolated sequence comprising a nucleotide sequence encoding the polypeptide         Producing the polypeptide activity in vivo         Administered to the subject in such a form,     Comprising   (ii) the subject needs to suppress the activity or expression of the polypeptide of claim 1;     If       (a) subject to a therapeutically effective amount of an antagonist to the polypeptide         And / or       (b) a nucleus that suppresses expression of a nucleotide sequence encoding the polypeptide         Administering an acid molecule to the subject; and / or       (c) for the polypeptide and its ligand, substrate, or receptor;         Administering to the subject a competing therapeutically effective amount of the polypeptide;     A method for treating the subject, comprising: 5. Associated with the expression or activity of the polypeptide of claim 1 in a subject.   A method of diagnosing a disease or susceptibility to the disease in the subject,   (a) in the nucleotide sequence encoding the polypeptide in the subject's genome     Determining whether the mutation is present and / or   (b) determining the presence or amount of expression of the polypeptide in a sample obtained from the subject.     Analyzing,   A method comprising: 6. Identify a compound that stimulates or suppresses the function of the polypeptide of claim 1.   Screening method for   (a) a candidate compound and the polypeptide (or a cell carrying the polypeptide);     Cell or its membrane) or a fusion protein thereof with the candidate compound.     Measuring with a label directly or indirectly attached to   (b) a candidate compound and the polypeptide (or a cell carrying the polypeptide);     Cell or its membrane) or its fusion protein     Measuring in the presence of   (c) a signal generated by the candidate compound upon activation or suppression of the polypeptide.     Whether or not it is appropriate for the cell or cell membrane carrying the polypeptide     Using a detection system   (d) combining the candidate compound and the solution containing the polypeptide to form a mixture.     And measuring the activity of the polypeptide in the mixture to determine the activity of the mixture.     Comparing with the standard, and   (e) the candidate compound is a mRNA encoding the polypeptide in a cell;     Detecting effects on polypeptide production, for example, using an ELISA assay     thing,   A screening method comprising a method selected from the group consisting of: 7. An agonist or antagonist of the polypeptide of claim 1. 8. The polypep according to claim 1, wherein the following expression system is present in a compatible host cell.   An expression system containing a polynucleotide capable of producing a tide. 9. The host cell is capable of growing the amino acid sequence of SEQ ID NO:   No amino acid sequence having at least 70% identity to the amino acid sequence.   A cell is formed using the expression system of claim 8 so as to produce a polypeptide.   Transforming or transfecting the recombinant host cell.   Production method. 10. A recombinant host cell obtained by the method according to claim 9. 11. At least for the amino acid sequence of SEQ ID NO: 2 over the entire length of SEQ ID NO: 2   Expressing a polypeptide comprising an amino acid sequence having 70% identity   The membrane of the recombinant host cell according to claim 10. 12. 11. A host cell according to claim 10 sufficient to produce said polypeptide.   Culturing under the conditions, and recovering the polypeptide from the culture medium.   A method for producing the polypeptide. 13. An isolated polynucleotide selected from the group consisting of the following (a) to (d):   (a) less than the nucleotide sequence of SEQ ID NO: 3 over the entire length of SEQ ID NO: 3     Nucleotides with 70%, 80%, 90%, 95% and 97% identity     An isolated polynucleotide comprising a nucleotide sequence,   (b) an isolated polynucleotide comprising the polynucleotide of SEQ ID NO: 3     ,   (c) the polynucleotide of SEQ ID NO: 3, or   (d) at least the amino acid sequence of SEQ ID NO: 4 over the entire length of SEQ ID NO: 4     Polypep with 70%, 80%, 90%, 95%, 97-99% identity     An isolated polynucleotide comprising a nucleotide sequence encoding a tide     De. 14． A polypeptide selected from the group consisting of (a) to (e):   (a) at least the amino acid sequence of SEQ ID NO: 4 over the entire length of SEQ ID NO: 4     Amino acids with 70%, 80%, 90%, 95%, 97-99% identity     A polypeptide comprising the sequence,   (b) at least the amino acid sequence of SEQ ID NO: 4 over the entire length of SEQ ID NO: 4     Amino acids with 70%, 80%, 90%, 95%, 97-99% identity   A polypeptide having a sequence, (c) a polypeptide comprising the amino acid of SEQ ID NO: 4, (d) a polypeptide consisting of the polypeptide of SEQ ID NO: 4, (e) encoded by a polynucleotide comprising the sequence contained in SEQ ID NO: 3   Polypeptide.