MXPA99010596A - Mu-1, member of the cytokine receptor family - Google Patents

Abstract

Polynucleotides encoding the MU-1 hematopoietin receptor superfamily chain and fragments thereof are disclosed. MU-1 proteins and methods for their production are also disclosed.

Description

U-1, MEMBER OF THE CYTOKINE RECEPTOR FAMILY Field of the Invention The present invention relates to new members of the mammalian cytokine receptor protein family (including, without limitation, human and murine receptor proteins) , fragments thereof and recombinant polynucleotides and cells useful for the expression of such proteins.

Background of 9th Invention A variety of regulatory molecules, known as hematopoietins, have been identified as being involved in the development and proliferation of the various populations of hematopoietic or blood cells. Most hematopoietins exhibit certain biological activities by interacting with a receptor on the target cell surface. The cytokine receptors are commonly composed of one, two or three chains. Many of the cytokine receptors and some cytokines, such as p40 of IL-12, are members of the hematopoietin receptor protein superfamily. The identification of new members of the superfamily of hematopoietin receptors can be useful in the regulation of hematopoiesis, in the regulation of immune responses and in the identification of other members of the hematopoietin superfamily, including cytokines and receptors. It would be desirable to identify and determine the DNA and protein sequence for the hitherto unknown members of the hematopoietin receptor superfamily.

OBJECTIVES OF THE INVENTION In accordance with the present invention, polynucleotides encoding the hematopoietin-MU-1 receptor superfamily chain are described, including, without limitation, those from murine and human sources.

In certain embodiments, the invention provides an isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: a) the nucleotide sequence of SEQ ID NO: 1; b) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 238 to nucleotide 1852; c) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 301 to nucleotide 1852; d) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 301 to nucleotide 945; e) a nucleotide sequence that varies from the nucleotide sequence specified in any of (a) - (d) as a result of a degeneracy of the genetic code; f) a nucleotide sequence capable of hybridizing under stringent conditions to the nucleotide specified in any of (a) - (d); g) a nucleotide sequence encoding a homologous species of the sequence of SEQ ID NO: 2; and h) an allelic variant of the nucleotide sequence specified in any of (a) - (d). Preferably, the nucleotide sequence codes for a protein having biological activity of the hematopoietin-MU-1 receptor superfamily chain. The nucleotide sequence can be operatively linked to an expression control sequence. The invention also provides isolated polynucleotides comprising a sequence of nucleotides encoding a peptide or protein comprising an amino acid sequence selected from the group consisting of: a) the amino acid sequence of SEQ ID NO: 2; b) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 538; c) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 236; d) the amino acid sequence of SEQ ID NO: 2, from amino acid 1 to amino acid 236; and e) fragments of (a) - (d) possessing biological activity of the hematopoietin-MU-1 superfamily of receptor chain. Host cells, preferably mammalian cells, transformed with the polynucleotides are also provided. In other embodiments, the invention also provides a process for the production of a MU-1 protein. The process comprises: a) growing a culture of the host cell of the present invention in an appropriate culture medium, and b) purifying the human MU-1 protein from the culture medium. Proteins produced according to these methods are also provided. The present invention also provides an isolated MU-1 protein, which comprises an amino acid sequence selected from the group consisting of: a) the amino acid sequence of SEQ ID NO: 2; b) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 538; c) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 236; d) the amino acid sequence of SEQ ID NO: 2, from amino acid 1 to amino acid 236; and e) fragments of (a) - (d) possessing biological activity of the hematopoietin-MU-1 superfamily of receptor chain. In other preferred embodiments, the specified amino acid sequence is part of a fusion protein (with an additional amino acid sequence not derived from MU-1). Preferred fusion proteins comprise an antibody fragment, such as an Fc fragment. Also provided are pharmaceutical compositions comprising a protein of the present invention and a pharmaceutically acceptable carrier.

The present invention further provides compositions comprising an antibody that specifically reacts with a protein of the present invention.

Detailed Description of the Preferred Modalities The inventors of the present application for the first time have identified and provided polynucleotides that encode the hematopoietin receptor superfamily chain MU-1 (hereinafter referred to as "MU-1" or " MU-1 protein "), including, without limitation, polynucleotides encoding human MU-1. A region of 70 amino acids of the human IL5 receptor (LMTNAFISIIDDLSKYDVQVRAAVSSMCREAGLWSEWSQPI GNDEHKPLREWFV IVIMATICFILLIL, SEQ ID NO: 3) was used to investigate the EST database of GenBank using the TBLASTN algorithm. The sequence within clone AC002303 of BAC from human chromosome 16p12 with homology to this region was identified, suggesting that this segment could encode a gene for a new hematopoietin receptor. Examination of open reading frames within lOQObp of 40,886 nucleotides revealed an open frame of 270bp that when used in a BLASTP search of GenPept exclusively identified members of the cytokine receptor family. A stop codon present at the end of this reading frame was interpreted as an indication of the transition over an exon / intron limit. It was then determined whether the RNA was transcribed from a gene contained within the BAC clone from chromosome 16p12. PCR primaries were synthesized based on the largest segment of ORF, which contained peptide sequence conserved within the family of cytokine receptors. The primers GAGTCCGAGGAGAAAGCTGATCTCA (5p) (SEQ ID NO: 4) and GAAAGATGACCGGGTCACTCCATT (3p) (SEQ ID NO: 5) were used in PCRs to screen phage libraries from various human tissues (Clontech). PCR products of the expected size of 164 bp that specifically hybridized to a 32-P labeled oligonucleotide of the sequence ACTCGAGCTATGAGCTGCAGGTGCGGGCA (SEQ ID NO: 6) were observed in phage from lung, kidney, placenta and heart. Using the oligonucleotide ACTCGAGCTATGAGCTGCAGGTGCGGGCA (SEQ ID NO: 7) a clone NN14-1b (MU-1) of full-length cDNA was identified, purified and sequenced. The DNA sequence and the predicted amino acid sequence are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively. The open reading frame codes for a new member of the hematopoietin receptor family. It has a leader sequence, conserved cysteine pairs, PP, and WSXWS motif characteristics (SEQ ID NO: 8) of the family, as well as a transmembrane domain and broad cytoplasmic domain. The subsequent alignment of FASTA of this sequence with GenPept showed the highest homology with human IL-2Rb. The predicted amino acid sequence of the receptor chain includes a putative signal sequence of amino acids 1-21. It is believed that mature human MU-1 has the amino acid sequence 24-538 of SEQ ID NO: 2. A transmembrane domain was found at amino acids 237-254. MU-1 cDNA was deposited with the American Type Culture Collection on March 10, 1998, under accession number ATCC 98687. Any forms of MU-1 proteins of less than full length are encompassed within the present invention and they are referred to here collectively with full length and mature forms such as "MU-1" or "MU-1 proteins". MU-1 proteins of less than full length can be produced by expression of a corresponding fragment of the polynucleotide encoding the full-length MU-1 protein (SEQ ID NO: 4 or SEQ ID NO: 6). These corresponding polynucleotide fragments are also part of the present invention. Modified polynucleotides as described above can be prepared by standard techniques of molecular biology, including the construction of appropriate mutants with the desired deletion, site-directed mutagenesis methods or by polymerase chain reaction using appropriate oligonucleotide primers.

For the purposes of the invention, a protein has "a biological activity of the hematopoietin-MU-1 receptor superfamily chain" if it possesses one or more of the biological activities of the corresponding mature MU-1 protein. MU-1 or active fragments thereof (MU-1 proteins) can also be fused to carrier molecules such as immunoglobulins. For example, soluble forms of MU-1 can be fused via "linker" sequences to the Fc portion of an immunoglobulin. Other fusion proteins, such as those with GST, Lex-A or MBP, can also be used. The invention also encompasses allelic variants of the nucleotide sequence as set forth in SEQ ID NO: 1, that is, naturally occurring alternative forms of the isolated polynucleotide of SEQ ID NO: 1 which also encode MU-1 proteins, preferably those proteins that have a biological activity of MU-1. Also included in the invention are isolated polynucleotides that hybridize to the nucleotide sequence set forth in SEQ ID NO: 1 under highly stringent conditions (eg, O.lxSSC at 65 ° C). Also included in the present invention are isolated polynucleotides which encode MU-1 proteins but which differ from the nucleotide sequence set forth in SEQ ID NO: 1 by virtue of the degeneracy of the genetic code. Also, variants of the nucleotide sequence as set forth in SEQ ID NO: 1 and which are caused by point mutations or by induced modifications are comprised in the present invention. The present invention also provides polynucleotides that encode homologs of human MU-1 from other animal species, particularly other mammalian species. The homologous species can be identified and isolated by preparing probes or primaries from the murine or human sequences described herein and screening a library from an appropriate species, such as, for example, libraries constructed from PBMCs, thymus or testes of species Relevant The isolated polynucleotides of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors described in Kaufman et al., Nucleic Acids Res. 19, 4485-4490 (1991), for the purpose of produce recombinantly the MU-1 protein. Many suitable expression control sequences are known in the art. General methods of expression of recombinant proteins are also known and are exemplified in R. Kaufman, Methods in Enzymology 185, 537-566 (1990). As defined herein "operably linked" means enzymatically or chemically linked to form a covalent bond between the isolated polynucleotide of the invention and the expression control sequence, in such a way that the MU-1 protein is expressed by a host cell that has been transformed (transfected) with the linked polynucleotide / expression control sequence. A number of cell types can act as host cells suitable for the expression of MU-1 protein. Any type of cell capable of expressing functional MU-1 protein can be used. Suitable mammalian host cells include, for example, monkey COS cells, Chinese hamster's ovary (CHO) cells, human kidney 293 cells, human A431 epidermal cells, human Colo205 cells, 3T3 cells, CV-1 cells , other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro cultures of primary tissue, primary explants, HeLa cells, mouse L cells, BHK cells, HL-60, U937, HaK Rat2, BaF3, 32D , FDCP-1, PC12, M1x or C2C12 cells. The MU-1 protein can also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system. Materials and methods for insect / virus cell expression systems are commercially available in the form of equipment in, for example, Invitrogen, San Diego, California, E.U.A. (the MaxBac® kit) and such methods are well known in the art, as described in Summers and Smith, Texas Aqricultural Experiment Station Bulletin No. 1555 (1987) which is incorporated herein by reference. MU-1 protein can also be produced in insect cells using appropriate isolated polynucleotides as described above Alternatively, the MU-1 protein can be produced in lower eukaryotes such as yeast or in prokaryotes such as bacteria Suitable strains of yeast include strains Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces, Candida, or any yeast strain capable of expressing heterologous proteins Suitable strains of bacteria include Escherichia coli, Bacillus subtilis, Salmonella typhimurium or any bacterial strain capable of expressing heterologous proteins. bacteria can result in the formation of inclusion bodies that i Incorporate the recombinant protein. Thus, refolding of the recombinant protein may be required in order to produce active or more active material. Various methods are well known in the art to correctly obtain folded heterologous proteins from bacterial inclusion bodies. These methods generally involve the solubilization of the protein from the inclusion bodies, subsequently denaturing the protein completely using a chaotropic agent. When cysteine residues are present in the primary amino acid sequence of the protein, it is often necessary to perform the retraction in an environment that allows the correct formation of disulfide bonds (a redox system). General methods of refolding are described in Kohno, Meth. Enzvm .. 185: 187-195 (1990). European patent EP 0433225 and the co-pending application of US Pat. Nos. 08 / 163,877 describe suitable methods. The MU-1 protein of the invention can also be expressed as a product of transgenic animals, v. gr., as a component of the milk of transgenic cows, goats, pigs or sheep that are characterized by germ or somatic cells containing a polynucleotide sequence that encodes the MU-1 protein. The MU-1 protein of the invention can be prepared by growing a culture of transformed host cells under culture conditions necessary to express the desired protein. The resulting expressed protein can then be purified from the culture medium or cell extracts. Soluble forms of the MU-1 protein of the invention can be purified from conditioned media. Membrane binding forms of the MU-1 protein of the invention can be purified by preparing a total membrane fraction from the expression cell and extracting the membranes with a nonionic surfactant such as Triton X-100. The MU-1 protein can be purified using methods known to those skilled in the art. For example, the MU-1 protein of the invention can be concentrated using a commercially available protein concentration filter, for example, a Millipore Pellicon or Amicon ultrafiltration unit. Following the concentration step, the concentrate can be applied to a purification matrix such as a gel filtration medium. Alternatively, an ion exchange resin, for example, a matrix or a substrate having diethylaminoethyl (DEAE) or polyethylene imine (PEI) groups may be employed. The matrices can be acrylamide, agarose, dextran, cellulose or other types commonly used in protein purification. Alternatively, a cation exchange step may be employed. Suitable cation exchangers include various insoluble matrices comprising sulfopropyl or carboxymethyl groups. Sulfopropyl groups are preferred (for example, S-Sepharose® columns). Purification of the MU-1 protein from the culture supernatant can also include one or more column steps on affinity resins such as concavalin A-agarose, heparin-toyopearl®, or Cibacrom blue 3GA Sepharose®, or by interaction chromatography hydrophobic using such resins as phenyl ether, butyl ether or propyl ether; or by immunoaffinity chromatography. Finally, one or more steps of high resolution reverse phase liquid chromatography (RP-HPLC) employing hydrophobic RP-HPLC media, for example, silica gel having suspended methyl or other aliphatic groups, can be used to further purify the MU-1 protein. Affinity columns including antibodies to the MU-1 protein can also be used in the purification according to known methods. Some or all of the preceding purification steps, in various combinations or with other known methods, may also be employed to provide an isolated and substantially purified recombinant protein. Preferably, the isolated MU-1 protein is purified so that it is substantially free of other mammalian proteins. The MU-1 proteins of the invention can also be used to screen agents that are capable of binding to MU-1. Binding assays using a desired binding protein, immobilized or not, are well known in the art and can be used for this purpose using the MU-1 protein of the invention. Screening assays based on purified cells or proteins (cell-free) can be used to identify such agents. For example, the MU-1 protein can be immobilized in purified form on a carrier and the binding or potential ligands for purified MU-1 protein can be measured. MU-1 proteins, purified from cells or recombinantly produced, can be used as a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Said composition may contain, in addition to the MU-1 or inhibitor and carrier, various diluents, fillers, salts, regulators, stabilizers, solubilizers, and other materials that are well known in the art. The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient (s). The characteristics of the carrier will depend on the administration route. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6. , IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-14, IL-15, G-CSF, progenitor cell factor and erythropoietin. The pharmaceutical composition may also include anti-cytokine antibodies. The pharmaceutical composition may contain thrombolytic or anti-thrombotic factors such as plasminogen activator and HIV Factor. The pharmaceutical composition may also contain other anti-inflammatory agents. Such additional factors and / or agents may be included in the pharmaceutical composition to produce a synergistic effect with the isolated MU-1 protein, or to minimize the side effects caused by the isolated MU-1 protein. Conversely, the isolated MU-1 protein can be included in formulations of the particular cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent to minimize side effects of the cytokine, lymphokine, another factor hematopoietic, thrombolytic or antithrombotic factor, or anti-inflammatory agent. The pharmaceutical composition of the invention may be in the form of a liposome in which the isolated MU-1 protein is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids that exist in aggregate form such as miscella, insoluble monolayers, liquid crystals, or laminar layers that are in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponins, bile acids, and the like. The preparation of such liposomal formulations is within the skill of the art, as described, for example, in U.S. Pat. No. 4,235,871; U.S. Patent A. No. 4,501,728; US Patent No. 4,837,028 and U.S. Pat. No. 4,737, 323; all of which is incorporated herein by reference. As used herein, the term "therapeutically effective amount" means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a significant benefit to the patient, eg, improvement of symptoms, cure or increase in curing speed of said conditions. When an active ingredient is applied to an individual, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. In the practice of the method of treatment or use of the present invention, a therapeutically effective amount of the isolated MU-1 protein is administered to a mammal. The isolated MU-1 protein can be administered according to the method of the invention, either alone or in combination with other therapies such as treatments employing cytokines, lymphokines or other hematopoietic factors. When co-administered with one or more cytokines, lymphokines or other hematopoietic factors, the MU-1 protein can be administered either simultaneously with cytokine (s), lymphokine (s), other hematopoietic factor (s) (s). ), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the treating physician will decide on the appropriate sequence of MU-1 protein administration in combination with cytokine (s), lymphokine (s), other hematopoietic factor (s), thrombolytic factors or anti-thrombotic. Administration of the MU-1 protein used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional forms, such as oral ingestion, inhalation, cutaneous injection, subcutaneous or intravenous Intravenous administration to the patient is the most preferred. When a therapeutically effective amount of the MU-1 protein is administered orally, the MU-1 protein will be in the form of a tablet, capsule, powder, solution or elixir. When administered in tablet form, the pharmaceutical composition of the invention may additionally contain a solid carrier such as gelatin or an adjuvant. The tablet, capsule and powder contains from about 5 to 95% of the MU-1 protein, and preferably from about 25 to 90% of the MU-1 protein. When administered in liquid form, a liquid carrier such as water, petroleum, animal or vegetable oils such as peanut oil, mineral oil, soybean oil, sesame oil, or synthetic oils can be added. The liquid form of the pharmaceutical composition may further contain physiological saline, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains from about 0.5 to 90% by weight of the MU-1 protein, and preferably from about 1 to 50% of MU-1 protein.

When a therapeutically effective amount of the MU-1 protein is administered by intravenous, cutaneous or subcutaneous injection, the MU-1 protein will be in the form of a parenterally acceptable and pyrogen-free aqueous solution. It is within the skill of the art to prepare such parenterally acceptable protein solutions, with due regard to pH, isotonicity, stability and the like. A pharmaceutically preferred composition for intravenous, cutaneous or subcutaneous injection must contain, in addition to the MU-1 protein, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose Injection and Sodium Chloride, Injection of Ringer with Lactate, or other vehicle as is known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, regulators, antioxidants, or other additives known to those skilled in the art. The amount of MU-1 protein in the pharmaceutical composition of the present invention will depend on the nature and severity of the condition being treated, and on the nature of previous treatments the patient has had. Finally, the attending physician will decide the amount of MU-1 protein with which he will treat each individual patient. Initially, the attending physician will administer low doses of the MU-1 protein and observe the patient's response. Higher doses of the MU-1 protein can be administered until the optimal therapeutic effect is obtained for the patient, and at that point generally the dose will not increase further. It is contemplated that the various pharmaceutical compositions used in the practice of the method of the present invention should contain about 0.1 μg to about 100 mg of the MU-1 protein per kilogram of body weight. The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary depending on the severity of the disease being treated and the potential idiosyncratic condition and response of each individual patient. It is contemplated that the duration of each application of the MU-1 protein will be in the range of 12 to 24 hours of continuous intravenous administration.

Finally the attending physician will decide about the appropriate duration of the intravenous therapy using the pharmaceutical composition of the present invention. It is expected that the polynucleotide and proteins of the present invention exhibit one or more of the biological uses or activities (including those associated with assays cited herein) that are identified below. The uses or activities described for the proteins of the present invention can be provided by the administration or use of such proteins or by the administration or use of polynucleotides that encode such proteins (such as, for example, in appropriate gene or vector therapies for the introduction of DNA).

Cytokine and Proliferation / Cell Differentiation Activity A protein of the present invention can exhibit cytokine activity, cell proliferation (either induction or inhibition) or cell differentiation (either induction or inhibition), or it can induce production of other cytokines in certain cell populations. Many protein factors discovered to date, including all known cytokines, have exhibited activity in one or more factor-dependent cell proliferation assays, and hence the assays serve as a convenient confirmation of cytokine activity. The activity of a protein of the present invention is evidenced by one of a number of routine factor-dependent cell proliferation assays for cell lines including, without limitation, 32D, DA2, DA1G, T10, B9, B9 / 11, BaF3 , MC9 / G, M + (preB M +), 2E8, RB5, DA1, 123, T1165, HT2, CTLL2, TF-1, Mo7e and CMK The activity of the protein of the invention can, among other means, be measured by the following methods: Assays for proliferation of thymocytes or T cells include, without limitation, those described in Current Protocols in Immunology, Ed. By J.E. Coligan, A.M. Kruisbeek, D.H. Marguiles, E.M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley Interscience (Chapter 3, In Vitro Assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic Studies in Humans); Takai et al., J.

Immunol. 137: 3494-3500, 1986; Bertagnolli et al., J. Immunol. 145: 1706-1712, 1990; Bertagnolli et al., Cellular Immunology 133: 327-341, 1991; Bertagnolli, et al., J. Immunol. 149: 3778-3783, 1992; Bowman et al., J. Immunol. 152: 1756-1761, 1994. Assays for determining production and / or proliferation of spleen cells, lymph node cells or thymocytes include, without limitation, those described in: Polyclonal T cell stimulation, Kruisbeek, A.M. and Shevach. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol. 1 pp. 3.12.1-3.12.14, John Wiley and Sons, Toronto, 1994; and Measurements of mouse and human Interferon ?, Schreiber, R.D. In Current Protocols in Immunology, J.E.e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toronto, 1994. Trials to determine the proliferation and differentiation of hematopoietic and lymphopoietic cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomly, K., Davis, LS and Lipsky, P.E. In Current Protocols in Immunology. J.E.e.a Coligan eds. Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toronto. 1991; deVries et al., J. Exp. Med. 173: 1205-1211, 1991; Moreau et al., Nature 336: 690-692, 1988; Greenberger et al., Proc. Nati Acad. Sci. U.S.A. 80: 2931-2938, 1983; Measurement of mouse and human interleukin 6 - Nordan, R. In Current Protocols in Immunology. J.E.e.a Coligan eds. Vol 1 pp. 6.6.1-6.6.5, John Wiley and Sons, Toronto. 1991; Smith et al., Proc. Nati Acad. Sci. U.S.A. 83: 1857-1861, 1986; Measurement of human Interleukin 11-Bennett, F. Giannotti, J. Clark, S.C. and Turner, K.J. In Current Protocols in Immunology. J.E.e.a Coligan eds. Vol 1 pp. 6.15.1, John Wiley and Sons, Toronto. 1991. Measurement of mouse and human Interleukin 9 - Ciarletta, A. Giannotti, J., Clark, S.C. and Turner, K.J. In Current Protocols in Immunology. J.E.e.a Coligan eds. Vol 1 pp. 6.13.1, John Wiley and Sons, Toronto. 1991. Assays to determine the responses of T cell clones to antigens (which will identify, among others, proteins that affect the interactions of APC-T cells as well as the effects of direct T cells by measuring proliferation and cytokine production) include , without limitation, those described in: Current Protocols ¡n Immunology, Ed by JE Coligan, A.M. Kruisbeek, D.H.

Margulies, E.M. Shevach, W. Stroper, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro Assays for Mouse Lymphocyte Function, Chapter 6, Cytokines and their cellular receptors, Chapter 7, Immupo.ogic studies in Humans); Weinberger et al., Proc. Nati Acad. Sci. U.S.A. 77: 6091-6095, 1980; Weinberger et al., Eur. J. Immun. 11: 405-411, 1981; Takai et al., J. Immunol. 137: 3494-3500, 1986; Takei et al., J. Immunol. 140: 508-512, 1988.

Immunological Stimulation or Suppression Activity A protein of the present invention may also exhibit immunological stimulation or immune suppression activity, including without limitation the activities for which assays have been described herein. A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SCID)), for example, in the regulation (up or down) of the growth and proliferation of T and / or B lymphocytes, as well as carrying out the cytolytic activity of NK cells and other cell populations. These immunological deficiencies can be genetic or viral (for example, HIV) as well as bacterial or fungal infections, or may result from autoimmune disorders. More specifically, infectious diseases caused by viruses, bacteria, fungi or other infection can be treated using a protein of the present invention, including infections by H1V, hepatitis viruses, herpes viruses, mycobacteria, lesmania spp, malaria spp and various infections by fungi such as candidiasis. Of course, in this regard, a protein of the present invention may also be useful where it is generally indicated to strengthen the immune system, for example, in the treatment of cancer. Autoimmune disorders that can be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythromatosis, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, diabetes mellitus dependent of insulin, myasthenia gravis, graft versus host disease and eye inflammation disease by autoimmunity. Said protein of the present invention may also be useful in the treatment of reactions and allergic conditions, such as asthma (particularly allergic asthma) and other respiratory problems. Other conditions, in which immunological suppression is desired (including, for example, organ transplantation) can also be treated using a protein of the present invention. MU-1 DNA also maps to the chromosomal site for Crohn's disease. As a result, proteins of the present invention can be employed to treat Crohn's disease and other inflammatory bowel diseases. The use of the proteins of the invention may also be possible for immune responses, in a number of ways. The hyporegulation may be in the form of inhibition or blocking of an immune response already in progress, or may involve the prevention of the induction of an immune response. The functions of activated T cells can be inhibited by suppressing T cell responses or by inducing specific tolerance in T cells, or both. The immunosuppression of T cell responses is generally an active, not antigen-specific process, which requires continuous exposure of the T cells to the suppressive agent. Tolerance, which involves the induction of non-response or anergy in T cells, is susceptible to distinction from immunosuppression because it is generally antigen-specific and persists after the exposure to the tolerance-creating agent has ended. Operationally, tolerance can be demonstrated by the lack of a T cell response under re-exposure to a specific antigen in the absence of the tolerance creating agent. The hyporegulation or prevention of one or more antigenic functions (including without limitation functions of B lymphocyte antigen (such as, for example, B7)), for example, preventing the synthesis of high level lymphokine by activated T cells, will be useful in situations of tissue, skin and organ transplants and in graft versus host disease (GVHD). For example, blocking T cell function should result in reduced destruction of tissue in tissue transplants. Typically, in tissue transplants, rejection of the transplant is initiated through recognition as foreign by the T cells, followed by an immune reaction that destroys the transplant. Administration of a molecule that inhibits or blocks the interaction of a B7 lymphocyte antigen with its natural ligand (s)) in immune cells (such as a soluble monomeric form of a peptide having B7-2 activity alone or in With a monomeric form of a peptide having an activity of another B-lymphocyte antigen (eg, B7-1, B7-3) or blocking antibody), before transplantation can lead to the binding of the molecule to the ) natural ligand (s) in the immune cells without the transmission of the corresponding co-stimulatory signal. Blocking the antigen function of the B lymphocyte in this case prevents the synthesis of cytokine by immune cells, such as T cells, and thus acts as an immunosuppressant. Moreover, the lack of co-stimulation may also be sufficient to anergize the T cells, thereby inducing tolerance in a subject. The induction of long-term tolerance by B lymphocyte antigen blocking reagents can avoid the need for repeated administration of these blocking reagents. To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of B lymphocyte antigens. The effectiveness of a particular blocking reagent in the prevention of a rejection of organ transplantation or GVHD can be evaluated using animal models that are capable of predicting efficacy in humans. Examples of suitable systems that may be used include allogeneic cardiac grafts in rats and islet cell grafts, xenogenic pancreatic grafts in mice, both of which have been used to examine the immunosuppressive effects of CTLA4lg fusion proteins in vivo as described in Lenschow et al. al., Science 257: 789-792 (1992) and Turka et al., Proc. Nati Acad. Sci. USA, 89: 11102-11105 (1992). In addition, murine models of GVHD (see Paul de., Fundamental Immunology, Raven Press, New York, 1989, pp. 846-847) can be used to determine the blocking effect of the B cell antigen function in vivo in the development of that disease.

Blocking the antigen function may also be therapeutically useful for the treatment of autoimmune diseases. Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against the same tissue and which promotes the production of cytokines and auto-antibodies involved in the pathology of diseases. The prevention of the activation of autoreactive T cells can reduce or eliminate symptoms of the disease. The administration of reagents that block the co-stimulation of T cells by breaking the receptor: ligand interactions of the B lymphocyte antigens can be used to inhibit the activation of T cells and prevent the production of autoantibodies or cytokines derived from T cells that may be involved in the unhealthy process. Additionally, blocking reagents can induce antigen-specific tolerance of autoreactive T cells that could lead to long-term relief of the disease. The efficacy of blocking reagents in the prevention or amelioration of autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include experimental murine autoimmune encephalitis, systemic lupus erythomatosis in MRL / lpr / lpr mice or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB rats, and experimental murine myasthenia gravis (see Paul of., Fundamental Immunoiogy, Raven Press, New York, 1989, pp. 840-856). The overregulation of an antigen function (preferably a B lymphocyte antigen function), as a means of over-regulating immune responses, may also be useful in therapies. Overregulation of immune responses may be in the form of improving an existing immune response or eliciting an initial immune response. For example, improving an immune response through the stimulation of the B-cell antigen function may be useful in cases of viral infection. In addition, systemic viral diseases such as influenza, the common cold and encephalitis could be alleviated by the systemic administration of stimulant forms of B lymphocyte antigens.

Alternatively, immune anti-viral responses can be improved in an infected patient by removing T cells from the patient, by co-stimulating T cells in vitro with APCs pulsed with viral antigen either by expressing a peptide of the present invention or in conjunction with a stimulant of a soluble peptide of the present invention and reintroducing the T cells activated in vitro within the patient. Another method for improving anti-viral immune responses would be to isolate infected cells from a patient, transfecting them with a nucleic acid encoding a protein of the present invention as described herein, so that the cells express all or a portion of the protein on its surface, and reintroduce the transfected cells into the patient. The infected cells would now be able to deliver a co-stimulatory signal to, and thus activate, the T cells in vivo. In another application, upregulation or amelioration of the antigen function (preferably B lymphocyte antigen function) could be useful in the induction of tumor immunity. Tumor cells (e.g., sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma) transfected with a nucleic acid encoding at least one peptide of the present invention can be administered to a subject to overcome the specific tolerance to the tumor in the subject. If desired, the tumor cells can be transfected to express a combination of peptides. For example, tumor cells obtained from a patient can be transfected ex vivo with an expression vector that directs the expression of a peptide having activity similar to B7-2., alone or in combination with a peptide having activity similar to B7-1 and / or activity similar to B7-3. The transfected tumor cells are returned to the patient to result in the expression of the peptides on the surface of the transfected cells. Alternatively, gene therapy techniques can be used to target a tumor cell for transfection in vivo. The presence of the peptide of the present invention, which has the activity of a B lymphocyte antigen (s) on the surface of the tumor cells, provides the necessary co-stimulation signal for the T cells to induce a immune response mediated by T cells against transfected tumor cells. In addition, tumor cells lacking MHC class I or MHC class II molecules, or that do not reexpress sufficient amounts of MHC class I or MHC class II molecules, can be transfected with nucleic acid encoding all or a portion of (for example, a truncated portion of the cytoplasmic domain) of an MHC class I chain protein and β2 microglobulin protein or an MHC class II chain protein and an MHC class γ chain protein to express this forms MHC class I or MHC class II proteins on the surface of the cell. Expression of the appropriate MHC class I or classes II, in conjunction with a peptide having the activity of a B lymphocyte antigen (eg, B7-1, B7-2, B7-.3) induces an immune response mediated by T cells against the transfected tumor cell. Optionally, a gene encoding an antisense construct that blocks the expression of an associated MHC class II protein, such as the constant chain, can also be cotransfected with a DNA encoding a peptide that has the activity of B lymphocyte antigen to promote the presentation of antigens associated with tumor and induce tumor-specific immunity. Thus, the induction of an immune response mediated by T cells in a human subject may be sufficient to overcome tumor-specific tolerance in the subject.

The activity of a protein of the invention can also, among other means, be measured by the following met: Suitable assays for determining cytotoxicity of splenocytes or thymocytes include, without limitation, those described in Current Protocols in Immunology, Edited by J.E. Coligan, A.M. Kruisbeek, D.H. Margul.es, E.M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro Assays for Mouse Lymphocyte Function 3.1.-3.19; Chapter 7, Immunologic Studies in Humans); Herrmann et al., Proc. Nati Acad. Sci. USA 78: 2488-2492, 1981; Herrmann et al., J. Immunol. 128: 1968-1974, 1982; Handa et al., J. Immunol. 135-1564-1572, 1985; Takai et al., J. Immunol. 137: 3494-3500, 1986; Takai et al., J. Immunol. 140: 508-512. 1988; Hermann et al., Proc. Nati Acad. Sci. USA 78: 2488-2492, 1981; Herrmann et al., J. Immunol. 128: 1968-1974, 1982; Handa et al., J. Immunol. 135: 1564-1572, 1985; Takai et al., J. Immunol. 137: 3494-3500, 1986; Bowman et al., J. Virology 61: 1992-1998; Takai et al., J. Immunol. 140: 508-512, 1988; Bertagnolli et al., Cellular Immunology 133: 327-341, 1991; Brown et al., J. Immunol. 153: 3079-3092, 1994. Trials to determine T cell-dependent immunoglobulin responses and isotype switching (which will identify, among others, proteins that modulate T-cell-dependent antibody responses and that affect Th1 / Th2 profiles. ) include, without limitation, those described in: Maliszewski, J. Immunol. 144: 3028-3033, 1990; and Assays for B cell function: In vitro antibody production, Mond, J.J. and Brunswick, M. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 3.8.1-3.8.16, John Wiley and Sons, Toronto. 1994. Mixed lymphocyte reaction (MLR) tests (which will identify, among others, proteins that predominantly generate Th1 and CTL responses) include, without limitation, those described in Current Protocols Immunology, Edited by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-lnterscience (Chapter 3, Sn Vitro Assays for Mouse Lymphocyte Function 3.1.-3.19; Chapter 7, Immunologic Studies in Humans); Takai et al., J. Immunol. 137: 3494-3500, 1986; Takai et al., J. Immunol. 140: 508-512, 1988; Bertagnolli et al., J. Immunol. 149: 3778-3783, 1992. Dendritic cell-dependent assays (which will identify, among others, proteins expressed by dendritic cells that activate natural T cells) include, without limitation, those described in: Guery et al., J. Immunol. 134: 536-544, 1995; Inaba et al., Journal of Experimental Medicine 173: 549-559, 1991; Macatonia et al., Journal of Immunology 154: 5071-5079, 1995; Porgador et al., Journal of Experimental Medicine 182: 255-260, 1995; Nair et al., Journal of Virology 67: 4062-4069, 1993; Huang et al., Science 264: 961-965, 1994; Macatonia et al., Journal of Experimental Medicine 169: 1255-1264, 1989; Bhardwaj et al., Journal of Clinical Investigation 94: 797-807, 1994; and Inaba et al., Journal of Experimental Medicine 172: 631-640, 1990.

The tests to determine the survival / apoptosis of lymphocytes (which will identify, among others, the proteins that prevent apoptosis after superantigenic induction and the proteins that regulate lymphocyte homeostasis) include, without limitation, those described in: Darzynkiewicz et al. al., Cytometry 13: 795-808, 1992; Gorczyca et al., Leukemia 7: 659-670, 1993 Gorczyca et al, Cancer Research 53: 1945-1951, 1993; Itoh et al., Cell 66: 233-243, 1991; Zacharchuk, Journal of Immunology 145: 4037-4045, 1990; Zamai et al., Cytometry 14: 891-897, 1993; Gorczyca et al., International Journal of Oncology 1: 639-648, 1992. Tests for proteins that influence early stages of development and confinement of T cells include, without limitation, those described in Antica et al., Blood 84,111-117 , 1994; Fine et al., Cellular Immunology 155: 111-122, 1994; Galy et al., Blood 85: 2770-2778, 1995; Toki et al., Proc. Nat. Acad. Sci. USA 88: 7548-7551, 1991.

Hematopoiesis Regulation Activity A protein of the present invention may be useful in the regulation of hematopoiesis and, consequently, in the treatment of deficiencies of myeloid or lymphoid cells. Even a marginal biological activity in support of colony-forming cells or factor-dependent cell lines indicates involvement in the regulation of hematopoiesis, that is, in supporting the growth and proliferation of erythroid progenitor cells alone or in combination with other cytokines. , indicating utility in this way, for example, in the treatment of various anemias or to be used in conjunction with irradiation / chemotherapy to stimulate the production of erythroid precursors and / or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes / macrophages (ie, traditional CSF activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelosuppression; in supporting the growth and proliferation of megakaryocytes and consequently of platelets, thereby allowing the prevention or treatment of various platelet disorders such as thrombocytopenia, and in general to be used in place or in a complementary manner to platelet transfusions; and / or in sustaining the growth and proliferation of hematopoietic progenitor cells, which are capable of maturing each and every one of the aforementioned hematopoietic cells and therefore find therapeutic utility in various progenitor cell disorders (such as those usually treated with transplants, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria) as well as in the repopulation of the division of progenitor cells that results from irradiation / chemotherapy, either in vivo or ex vivo (that is, in conjunction with transplantation of bone marrow or with the transplantation of peripheral progenitor cells (homologous or heterologous)) as normal cells or genetically engineered by gene therapy. The activity of a protein of the invention can, among other means, be measured by the following methods: Appropriate tests to determine the proliferation and differentiation of several hematopoietic lines have been cited above. Tests for the differentiation of embryonic progenitor cells (which will identify, among others, proteins that influence the hematopoiesis of embryonic differentiation) include, without limitation, those described in Johansson et al. Cellular Biology 15: 141-151, 1995; Keller et al., Molecular and Cellular Biology 13: 473-486, 1993; McCIanahan et al., Blood 81: 2903-2915, 1993. Tests to determine the survival and differentiation of progenitor cells (which will identify, among others, proteins that regulate lympho-hematopoiesis) include, without limitation, those described in: Methylcellulose colony forming assays, Freshney, MG In Culture of Hematopoietic Cells, R.l. Freshney, et al. Vol pp 265-268, Wiley-Liss, Inc., New York, NY 1994; Hirayama et al., Proc Nati. Acad. Sci. USA 89: 5907-5911, 1992; Primitive hematopoietic colony forming cells with high proliferative potential, McNiece, I.K. and Briddell, R.A. In Culture of Hematopoietic Cells. R.l. Freshney, et al. eds. Vol pp. 23-39, Wiley-Liss, Inc. New York, NY 1994; Neben et al., Experimental Hematology 22: 353-359, 1994; Cobblestone area forming cell assay, Ploemacher, R.E. In Culture of Hematopoietic Cells. R.I. Freshney et al., Eds. Vol pp. 1-21, Wiley-Liss, Inc. New York, NY 1994; Long term bone cultures in the presence of stromal cells, Spooncer, E., Dexter, M. and Alien, T. In Culture of Hematopoietic Cells. R.l. Freshney et al., Eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, NY 1994; Long term culture initiating cell assay, Sutherland, H.J. In Culture of Hematopietic Cells, R.l. Freshney et al., Eds. Vol pp. 139-162, Wiley-Liss, Inc. New York, NY. 1994 Uses in Research and Utilities The polynucleotides provided by the present invention can be used by the research community for various purposes. The polynucleotides can be used to express recombinant protein for analysis, characterization or therapeutic use; as markers for tissues in which the corresponding protein is expressed preferentially (either constitutively or in a particular stage of tissue differentiation or development or in diseased states); as molecular weight markers on Southern gels; as markers or flags of chromosomes (when marked) to identify chromosomes or for mapping of related gene positions; for comparison with endogenous DNA sequences in patients to identify potential genetic disorders; as probes for hybridization and thus discover new related DNA sequences; as a source of information to derive primary PCR for genetic screening; as a probe for "subtracting" known sequences in the process of discovering new polynucleotides; for the selection and labeling of oligomers for binding to a "genetic chip" or other support, including for the examination of expression patterns; to generate anti-protein antibodies using DNA immunization techniques; and as an antigen to generate anti-DNA antibodies or to stimulate another immune response: Where the polynucleotide codes for a protein that binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction) , the polynucleotide can also be used in interaction trap assays (such as, for example, that described in Guris et al., Cell 75: 791-803 (1993)) to identify polynucleotides that encode the other protein with which it occurs. the binding or to identify inhibitors of the binding interaction. The proteins provided by the present invention can similarly be used in assays to determine biological activity, including in a panel of multiple proteins for high-throughput screening; to generate antibodies or to produce another immune response; as a reagent (including marker reagent) in assays designed to quantitatively determine levels of the protein (or its receptor) in biological fluids; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or in a particular stage of tissue differentiation or development or in a diseased state); and, of course, to isolate correlative receptors or ligands. Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the other protein with which the binding occurs or to identify inhibitors of the union interaction. The proteins involved in these binding interactions can also be used to screen peptides or small inhibitors of molecules or agonists of the binding interaction. Any of these research utilities are capable of being developed in reactive grade or in game format (kit) for commercialization as research products. The methods for performing the uses listed above are well known to those skilled in the art. References describing such methods include, without limitation, "Molecular Cloning: A Laboratory Manual", 2pd ed. Cold Spring Harbor Laboratory Press, Sambrook, J., E.F. Fritsch and T. Maniatis eds., 1989, and "Methods in Enzymology: Guide to Molecular Cloning Techniques", Academic Press, Berger, S.L. walk. Kimmel eds., 1987.

Nutritional Uses The polynucleotides and proteins of the present invention can also be used as nutritional sources or supplements. Such uses include, without limitation, use as a supplement of amino acids or proteins, use as a carbon source, the use as a source of nitrogen and the use as a source of carbohydrates. In such cases the protein or polynucleotide of the invention can be added to the feed of a particular organism or it can be administered as a separate solid or liquid preparation, such as in the form of a powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the protein or polynucleotide of the invention can be added to the medium in or on which the microorganism is cultured. The MU-1 proteins of the invention can also be used to immunize animals to obtain polyclonal and monoclonal antibodies that specifically react with the MU-1 protein and which can inhibit binding of ligands to the receptor. Such antibodies can be obtained using either the complete MU-1 protein as an immunogen, or using fragments of MU-1. Small fragments of MU-1 can also be used to immunize animals. The peptide immunogens may additionally contain a cysteine residue at the carboxyl terminus, and be conjugated to a hapten such as keyhole limpet hemocyanin (KLH). Additional peptide immunogens can be generated by replacing tyrosine residues with tyrosine sulphated residues. Methods for synthesizing such peptides are known in the art, for example, as in R.P. Merrifield, J. Amer. Chem. Soc. 85, 2149-2154 (1963); J.L. Krstenansky, et al. FEBS Lett. 211, 10 (1987). Neutralization or non-neutralization antibodies (preferably monoclonal antibodies) that bind to the MU-1 protein may also be useful therapeutic agents for certain tumors and also in the treatment of conditions described above. These monoclonal neutralizing antibodies may be capable of blocking the binding of ligands to the MU-1 receptor chain. All of the patents and literature references cited herein are incorporated herein by reference as if they had been fully disclosed.

LIST OF SEQUENCES (1) INFORMATION INFORMATION: (i) APPLICANT: Genetics Institute, Inc. (ii) TITLE OF INVENTION: MU-1 RECEIVER (iii) NUMBER OF SEQUENCES: 8 (iv) ADDRESS FOR CORRESPONDENCE: (A) RECIPIENT : Genetics Institute, Inc. (B) STREET: 87 CambridgePark Drive (C) CITY: Cambridge (D) STATE: MA (E) COUNTRY: USA (F) ZIP CODE: 02140 (v) COMPUTER LEADABLE FORM: (A) TYPE OF MEDIUM: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) PROGRAM: Patentln Relay # 1.0, Version # 1.30 (vi) CURRENT DATA OF THE APPLICATION: ( A) NUMBER OF APPLICATION: (B) DATE OF SUBMISSION: (C) CLASSIFICATION: (viii) INFORMATION OF POWDER / AGENT: (A) NAME: Brown, Scott A (B) REGISTRATION NUMBER: 32,724 (C) NUMBER OF RECORD / REFERENCE: GI5320 (ix) INFORMATION FOR TELECOMMUNICATIONS: (A) TELEPHONE: 617-498-8224 (B) TELEFAX: 617-876-5851 (2) INFORMATION FOR SEQ ID NO: l: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 2665 base pairs (B) TYPE: nucleic acid (C) STRING: double (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: cDNA (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: l: GTCGACTGGA GGCCCAGCTG CCCGTCATCA GAGTGACAGG TCTTATGACA GCCTGATTGG 60 TGACTCGGGC TGGGTGTGGA TTCTCACCCC AGGCCTCTGC CTGCTTTCTC AGACCCTCAT 120 CTGTCACCCC CACGCTGAAC CCAGCTGCCA CCCCCAGAAG CCCATCAGAC TGCCCCCAGC 180 ACACGGAATG GATTTCTGAG ÁAGAAGCCG AAACAGAAGG CCCGTGGGAG TCAGCATGCC 240 GCGTGGCTGG GCCGCCCCCT TGCTCCTGCT GCTGCTCCAG GGAGGCTGGG GCTGCCCCGA 300 CCTCGTCTGC TACACCGATT ACCTCCAGAC GGTCATCTGC ATCCTGGAAA TGTGGAACCT 360 CCACCCCAGC ACGCTCACCC TTACCTGGCA AGACCAGTAT GAAGAGCTGA AGGACGAGGC 420 CACCTCCTGC AGCCTCCACA GGTCGGCCCA CAATGCCACG CATGCCACCT ACACCTGCCA 480 CATGGATGTA TTCCACTTCA TGGCCGACGA CATTTTCAGT GTCAACATCA CAGACCAGTC 540 TGGCAACTAC TCCCAGGAGT GTGGCAGCTT TCTCCTGGCT GAGAGCATCA AGCCGGCTCC 600 CCCTTTCAAC GTGACTGTGA CCTTCTCAGG ACAGTATAAT ATCTCCTGGC GCTCAGATTA 660 CGAAGACCCT GCCTTCTACA TGCTGAAGGG CAAGCTTCAG TATGAGCTGC AGTACAGGAA 720 CCGGGGAGAC CCCTGGGCTG TGAGTCCGAG GAGAAAGCTG ATCTCAGTGG ACTCAAGAAG 780 TGTCTCCCTC CTCCCCCTGG AGTTCCGCAA AGACTCGAGC TATGAGCTGC AGGTGCGGGC 840 AGGGCCCATG CCTGGCTCCT CCTACCAGGG GACCTGGAGT GAATGGAGTG ACCCGGTCAT 900 CTTTCAGACC CAGTCAGAGG AGTTAAAGGA AGGCTGGAAC CCTCACCTGC TGCTTCTCCT 960 CCTGCTTGTC ATAGTCTTCA TTCCTGCCTT CTGGAGCCTG AAGACCCATC CATTGTGGAG 1020 GCTATGGAAG AAGATATGGG CCGTCCCCAG CCCTGAGCGG TTCTTCATGC CCCTGTACAA 1080 GGGCTGCAGC GGAGACTTCA AGAAATGGGT GGGTGCACCC TTCACTGGCT CCAGCCTGGA 1140 GCTGGGACCC TGGAGCCCAG AGGTGCCCTC CACCCTGGAG GTGTACAGCT GCCACCCACC 1200 ACGGAGCCCG GCCAAGAGGC TGCAGCTCAC GGAGCTACAA GAACCAGCAG AGCTGGTGGA 1260 GTCTGACGGT GTGCCCAAGC CCAGCTTCTG GCCGACAGCC CAGAACTCGG GGGGCTCAGC '1320 TTACAGTGAG GAGAGGGATC GGCCATACGG CCTGGTGTCC ATTGACACAG TGACTGTGCT 1380 AGATGCAGAG GGGCCATGCA CCTGGCCCTG CAGCTGTGAG GATGACGGCT ACCCAGCCCT 1440 GGACCTGGAT GCTGGCCTGG AGCCCAGCCC AGGCCTAGAG GACCCACTCT TGGATGCAGG 1500 GACCACAGTC CTGTCCTGTG GCTGTGTCTC AGCTGGCAGC CCTGGGCTAG GAGGGCCCCT 1560 GGGAAGCCTC CTGGACAGAC TAAAGCCACC CCTTGCAGAT GGGGAGGACT GGGCTGGGGG 1620 ACTGCCCTGG GGTGGCCGGT CACCTGGAGG GGTCTCAGAG AGTGAGGCGG GCTCACCCCT 1680 GGCCGGCCTG GATATGGACA CGTTTGACAG TGGCTTTGTG GGCTCTGACT GCAGCAGCCC 1740 TGTGGAGTGT GACTTCACCA GCCCCGGGGA CGAAGGACCC CCCCGGAGCT ACCTCCGCCA 1800 GTGGGTGGTC ATTCCTCCGC CACTTTCGAG CCCTGGACCC CAGGCCAGCT AATGAGGCTG 1860 ACTGGATGTC CAGAGCTGGC CAGGCCACTG GGCCCTGAGC CAGAGACAAG GTCACCTGGG 1920 CTGTGATGTG AAGACACCTG CAGCCTTTGG TCTCCTGGAT GGGCCTTTGA GCCTGATGTT 1980 TACAGTGTCT GTGTGTGTGT GTGCATATGT GTGTGTGTGC ATATGCATGT GTGTGTGTGT 2040 GTGTGTCTTA GGTGCGCAGT GGCATGTCCA CGTGTGTGTG TGATTGCACG TGCCTGTGGG 2100 CCTGGGATAA TGCCCATGGT ACTCCATGCA TTCACCTGCC CTGTGCATGT CTGGACTCAC 2160 GGAGCTCACC CATGTGCACA AGTGTGCACA GTAAACGTGT TTGTGGTCAA CAGATGACAA 2220 CAGCCGTCCT CCCTCCTAGG GTCTTGTGTT GCAAGTTGGT CCACAGCATC TCCGGGGCTT 2280 TGTGGGATCA GGGCATTGCC TGTGACTGAG GCGGAGCCCA GCCCTCCAGC GTCTGCCTCC 2340 AGGAGCTGCA AGAAGTCCAT ATTGTTCCTT ATCACCTGCC AACAGGAAGC GAAAGGGGAT 2400 GGAGTGAGCC CATGGTGACC TCGGGAATGG CAATTTTTTG GGCGGCCCCT GGACGAAGGT 2460 CTGAATCCCG ACTCTGATAC CTTCTGGCTG TGCTACCTGA GCCAAGTCGC CTCCCCTCTC 2520 TGGGCTAGAG TTTCCTTATC CAGACAGTGG GGAAGGCATG ACACACCTGG GGGAAATTGG 2580 CGATGTCACC CGTGTACGGT ACGCAGCCCA GAGCAGACCC TCAATAAACG TCAGCTTCCT 2640 TCAAAAAAAA AAAAAAAAAT CTAGA 2665 (2) INFORMATION FOR SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 538 amino acids (B) TYPE: amino acid (C) CHAIN: (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 2: Met Pro Arg Gly Trp Wing Wing Pro Leu Leu Leu Leu Leu Gln Gly 1 5 10 15 Gly Trp Gly Cys Pro Asp Leu Val Cys Tyr Thr Asp Tyr Leu Gln Thr 20 25 30 Val lie Cys lie Leu Glu Met Trp Asn Leu His Pro Ser Thr Leu Thr 35 40 45 Leu Thr Trp Gln Asp Gln Tyr Glu Glu Leu Lys Asp Glu Ala Thr Ser 50 55 60 Cys Ser Leu His Arg Ser Ala Ala Asn Ala Thr His Ala Thr Tyr Thr 65 70 75 80 Cys His Met Asp Val Phe His Phe Met Wing Asp Asp lie Phe Ser Val 85 90 95 Asn Lie Thr Asp Gln Ser Gly Asn Tyr Ser Gln Glu Cys Gly Ser Phe 100 105 110 Leu Leu Ala Glu Ser lie Pro Lys Pro Pro Phe Asn Val Thr Val 115 120 125 Thr Phe Ser Gly Gln Tyr Asn Lie Ser Trp Arg Ser Asp Tyr Glu Asp 130 135 140 Pro Wing Phe Tyr Met Leu Lys Gly Lys Leu Gln Tyr Glu Leu Gln Tyr 145 150 155 160 Arg Asn Arg Gly Asp Pro Trp Wing Val Ser Pro Arg Arg Lys Leu lie 165 170 175 Ser Val Asp Ser Arg Ser Val Ser Leu Leu Pro Leu Glu Phe Arg Lys 180 185 190 Asp Ser Ser Tyr Glu Leu Gln Val Arg Wing Gly Pro Met Pro Gly Ser 195 200 205 Ser Tyr Gln Gly Thr Trp Ser Glu Trp Ser Asp Pro Val lie Phe Gln 210 215 220 Thr Gln Ser Glu Glu Leu Lys Glu Gly Trp Asn Pro His Leu Leu Leu 225 230 235 240 Leu Leu Leu Leu Val lie Val Phe lie Pro Ala Phe Trp Ser Leu Lys 245 250 255 Thr His Pro Leu Trp Arg Leu Trp Lys Lys Lie Trp Wing Val Pro Ser 260 265 270 Pro Glu Arg Phe Phe Met Pro Leu Tyr Lys Gly Cys Ser Gly Asp Phe 275 280 285 Lys Lys Trp Val Gly Wing Pro Phe Thr Gly Ser Leu Glu Leu Gly 290 295 300 Pro Trp Ser Pro Glu Val Pro Ser Thr Leu Glu Val Tyr Ser Cys His 305 310 315 320 Pro Pro Arg Ser Pro Wing Lys Arg Leu Gln Leu Thr Glu Leu Gln Glu 325 330 335 Pro Wing Glu Leu Val Glu Ser Asp Gly Val Pro Lys Pro Ser Phe Trp 340 345 350 Pro Thr Wing Gln Asn Ser Gly Gly Ser Wing Tyr Ser Glu Glu Arg Asp 355 360 365 Arg Pro Tyr Gly Leu Val Ser lie Asp Thr Val Thr Val Leu Asp Wing 370 375 380 Glu Gly Pro Cys Thr Trp Pro Cys Ser Cys Glu Asp Asp Gly Tyr Pro 385 390 395 400 Wing Leu Asp Leu Asp Wing Gly Leu Glu Pro Ser Pro Gly Leu Glu Asp 405 410 415 Pro Leu Leu Asp Wing Gly Thr Thr Val Leu Ser Cys Gly Cys Val Ser 420 425 430 Wing Gly Ser Pro Gly Leu Gly Gly Pro Leu Gly Ser Leu Leu Asp Arg 435 440 445 Leu Lys Pro Pro Leu Wing Asp Gly Glu Asp Trp Wing Gly Gly Leu Pro 450 455 460 Trp Gly Gly Arg Ser Pro Gly Gly Val Ser Glu Ser Glu Wing Gly Ser 465 470 475 480 Pro Leu Wing Gly Leu Asp Met Asp Thr Phe Asp Ser Gly Phe Val Gly 485 490 495 Ser Asp Cys Ser Ser Pro Val Glu Cys Asp Phe Thr Ser Pro Gly Asp 500 505 510 Glu Gly Pro Pro Arg Ser Tyr Leu Arg Gln Trp Val Val lie Pro Pro 515 520 525 Pro Leu Ser Ser Pro Gly Pro Gln Ala Ser 530 535 (2) INFORMATION FOR SEQ ID NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 70 amino acids (B) TYPE: amino acid (C) CHAIN: (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 3: Leu Met Thr Asn Ala Phe lie Ser lie lie Asp Asu Leu Ser Lys Tyr 1 5 10 15 Asp Val Gln Val Arg Ala Ala Val Ser Ser Met Cys Arg Glu Ala Gly 20 25 30 Leu Trp Ser Glu Trp Ser Gln Pro lie Tyr Val Gly Asn Asp Glu His 35 40 45 Lys Pro Leu Arg Glu Trp Phe Val lie Val lie Met Wing Thr lie Cys 50 55 60 Phe lie Leu Leu lie Leu 65 70 (2) INFORMATION FOR SEQ ID NO: 4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (A) DESCRIPTION: / desc = "oligonucleotide" '(xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 4: GAGTCCGAGG AGAAAGCTGA TCTCA 25 (2) INFORMATION FOR SEQ ID NO: 5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (A) DESCRIPTION: / desc = "oligonucleotide" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 5: GAAAGATGAC CGGGTCACTC CATT 24 (2) INFORMATION FOR SEQ ID NO: 6: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (A) DESCRIPTION: / desc = "oligonucleotide" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 6: ACTCGAGCTA TGAGCTGCAG GTGCGGGCA 29 (2) INFORMATION FOR SEQ ID NO: 7: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (A) DESCRIPTION: / desc = "oligonucleotide" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 7: ACTCGAGCTA TGAGCTGCAG GTGCGGGCA 29 (2) INFORMATION FOR SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) CHAIN: (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 8: Trp Ser Xaa Trp Ser 1 5

Claims (25)

1. Novelty of the Invention 1. An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: a) the nucleotide sequence of SEQ ID NO: 1; b) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 238 to nucleotide 1852; c) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 301 to nucleotide 1852; d) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 301 to nucleotide 945; e) a nucleotide sequence that varies from the nucleotide sequence specified in any of (a) - (d) as a result of a degeneracy of the genetic code; f) a nucleotide sequence capable of hybridizing under stringent conditions to the nucleotide specified in any of (a) - (d); g) a nucleotide sequence encoding a homologous species of the sequence of SEQ ID NO: 2; and h) an allelic variant of the nucleotide sequence specified in any of (a) - (d).
2. 2. The polynucleotide of claim 1, wherein said nucleotide sequence codes for a protein having a biological activity of the hematopoietin-MU-1 receptor superfamily chain.
3. 3. The polynucleotide of claim 1, wherein said nucleotide sequence is operably linked to an expression control sequence.
4. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO: 1.
5. 5. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO: 1, from nucleotide 238 to nucleotide 1852
6. 6. A host cell transformed with the polynucleotide of claim 3.
7. 7. The host cell of claim 6, wherein said cell is a mammalian cell.
8. 8. A process for producing a MU-1 protein, said process comprising the steps of: a) growing a culture of the host cell of claim 6 in an appropriate culture medium; and b) purifying the MU-1 protein from the culture.
9. 9. An isolated MU-1 protein comprising an amino acid sequence selected from the group consisting of: a) the amino acid sequence of SEQ ID NO: 2; b) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 538; c) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 236; d) the amino acid sequence of SEQ ID NO: 2, from amino acid 1 to amino acid 236; and e) fragments of (a) - (d) possessing biological activity of the hematopoietin-MU-1 superfamily of hemopoietin receptors.
10. The protein of claim 9, comprising the amino acid sequence of SEQ ID NO: 2.
11. 11. The protein of claim 9, comprising the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 538
12. 12. The protein of claim 9, comprising the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 236.
13. 13. The protein of claim 9, comprising the amino acid sequence of SEQ ID NO: 2, from amino acid 1 to amino acid 236.
14. 14. A pharmaceutical composition comprising a protein of claim 9 and a pharmaceutically acceptable carrier.
15. 15. A protein produced in accordance with the process of claim 8.
16. 16. A composition comprising an antibody that specifically reacts with a protein of claim 9.
17. 17. An isolated polynucleotide comprising a nucleotide sequence encoding a peptide or protein comprising an amino acid sequence selected from the group consisting of: a) the amino acid sequence of SEQ ID NO: 2; b) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 538; c) the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 236; d) the amino acid sequence of SEQ ID NO: 2, from amino acid 1 to amino acid 236; and e) fragments of (a) - (d) possessing biological activity of the hematopoietin-MU-1 superfamily of receptor chain.
18. 18. The protein of claim 9, wherein said amino acid sequence is part of a fusion protein.
19. 19. The protein of claim 18, comprising an Fc fragment.
20. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO: 1, from nucleotide 301 to nucleotide 1852.
21. 21. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO: 1 , from nucleotide 238 to nucleotide 945.
22. 22. The polynucleotide of claim 17, which codes for a peptide or protein comprising the amino acid sequence of SEQ ID NO: 2.
23. 23. The polynucleotide of claim 17, which encodes a peptide or protein comprising the amino acid sequence of SEQ ID NO: 2, from amino acid 22 to amino acid 538.
24. 24. The polynucleotide of claim 17, which encodes a peptide or protein comprising the sequence of amino acids of SEQ ID NO: 2, from amino acid 22 to amino acid 236.
25. 25. The polynucleotide of claim 17, which encodes a peptide or protein comprising the amino acid sequence of SEQ ID NO: 2, from amino acid 1 to amino acid 236.