HUMAN INTERLEUKIN-11 RECEPTOR

Abstract: Polynucleotides encoding the human IL-11 receptor and fragments thereof are disclosed. IL-11 receptor proteins, methods for their production, inhibitors of binding of human IL-11 and its receptor and methods for their identification are also disclosed.

Description: HUMAN INTERLEUKEV-ll RECEPTOR Field of the Invention The present invention relates to the human interleukin-11 receptor, fragments thereof and recombinant polynucleotides and cells useful for expressing such proteins. Background of the Invention A variety of regulatory molecules, known as cytokines, have been identified including interleukin-11 (IL-11). The various protein forms of IL-11 and DNA encoding various forms of IL-11 activity are described in Bennett, et al, USPN 5,215,895 (June 1, 1993); McCoy, et ai, USPN 5,270,181 (December 14, 1993); and McCoy, et ai , USPN 5.292,646 (March 8, 1994), all incorporated herein by reference. Thus, the term "IL-11 " includes proteins having the biological activity described in these patents, whether produced by recombinant genetic engineering techniques; purified from cell sources producing the factor naturally or upon induction with other factors;' or synthesized by chemical techniques; or a combination of the foregoing. IL-11 is a pleiotropic cytokine that has been implicated in production of several biological activities including: induction of multipotential hematopoietic progenitor cell proliferation (Musashi et al. (1991) Blood. 78, 1448- 1451); enhancement of megakaryocyte and platelet formation (Burstein ei al. ( 1992) J Cell. Physiol. , 153, 305-312); stimulation oϊ acute phase protein synthesis (Baumann et al. (1991) J. Biol. Chem. , 266, 20424-20427); inhibition of adipocyte lipoprotein lipase activity (Kawashima et al. (1991) FEBS Lett. , 283, 199-202); and effects on neurotransmitter phenotype (Farm et al. (1994) Proc. Natl. Acad. Sci. USA, 91 , 43-47). IL-11 may be used in a pharmaceutical preparation or formulation to treat immune deficiencies, specifically deficiencies in hematopoietic progenitor cells, or disorders relating thereto. Treatment of the other disorders or stimulation of the immune systems of cells thereof may also employ IL-11. IL- 11 may also be employed in methods for treating cancer and other disease. Such pathological states may result from disease, exposure to radiation or drugs, and include, for example, leukopenia, bacterial and viral infections, anemia, B cell or T cell deficiencies such as immune cell or hematopoietic cell deficiency following a bone marrow transplantation. IL-11 may also be used to potentiate the immune response to a variety of vaccines creating longer lasting and more effective immunity. Therapeutic treatment of cancer and other diseases with IL-11 may avoid undesirable side effects caused by treatment with presently available drugs. Like most cytokines, IL-11 exhibits certain biological activities by interacting with an IL-11 receptor (IL-11R) on the surface of target cells. It would be desirable to identify and clone the sequence for the human receptor so that IL-1 1R proteins can be produced for various reasons, including production of therapeutics and screening for inhibitors of IL- 1 1 binding to the receptor and receptor signalling. Summary of the Invention In accordance with the present invention, polynucleotides encoding the human interleukin-11 receptor are disclosed. 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 from nucleotide 803 to nucleotide 1999; (b) a nucleotide sequence varying from the sequence of the nucleotide sequence specified in (a) as a result of degeneracy of the genetic code; and (c) an allelic variant of the nucleotide sequence specified in (a). Preferably, the nucleotide sequence encodes a protein having a biological activity of the human IL-11 receptor. The nucleotide sequence may be operably linked to an expression control sequence. In preferred embodiments, the polynucleotide comprises me nucleotide sequence of SEQ ID NO: l from nucleotide 803 to nucleotide 1999 or a fragment thereof; the nucleotide sequence of SEQ ID NO: l from nucleotide 803 to nucleotide 1828 or a fragment thereof; the nucleotide sequence of SEQ ID NO: l from nucleotide 1904 to nucleotide 1999 or a fragment thereof; the nucleotide sequence of SEQ ID NO: l from nucleotide 734 to nucleotide 1999 or a fragment thereof; the nucleotide sequence of SEQ ID NO. l from nucleotide 1067 to nucleotide 1828 or a fragment thereof; or the nucleotide sequence of SEQ ID NO: l from nucleotide 1067 to nucleotide 1999 or a fragment thereof. In other embodiments, the polynucleotide comprises a nucleotide sequence capable of hybridizing to the nucleotides sequence of SEQ ID NO: 1 under highly stringent conditions. The invention also provides isolated polynucleotides 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 acids 24 to 422; (c) the amino acid sequence of SEQ ID NO: 2 from amino acids 24 to 365; (d) the amino acid sequence of SEQ ID NO:2 from amino acids 391 to 422; (e) the amino acid sequence of SEQ ID NO:2 from amino acids 112 to 422; (f) the amino acid sequence of SEQ ID NO:2 from amino acids 112 to 365; and (g) fragments of (a)-(f) having a biological activity of the human IL- 11 receptor. Host cells, preferably mammalian cells, transformed with the polynucleotides are also provided. In other embodiments, the invention provides a process for producing a human IL-11R protein. The process comprises: (a) growing a culture of the host cell of the present invention in a suitable culture medium; and (b) purifying the human IL-11R protein from the culture. Proteins produced according to these methods are also provided. The present invention also provides for an isolated human IL-11R 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 acids 24 to 422; (c) the amino acid sequence of SEQ ID NO: 2 from amino acids 24 to 365; (d) the amino acid sequence of SEQ ID NO:2 from amino acids 391 to 422; (e) the amino acid sequence of SEQ ID NO: 2 from amino acids 112 to 422; (f) the amino acid sequence of SEQ ID NO:2 from amino acids 112 to 365; and (g) fragments of (a)-(f) having a biological activity of the human IL- 11 receptor. Preferably the protein comprises the amino acid sequence of SEQ ID NO: 2; the sequence from amino acid 24 to 422 of SEQ ID NO: 2; the sequence from amino acid 24 to 365 of SEQ ID NO:2; or the sequence from amino acid 391 to 422 of SEQ ID NO:2. Pharmaceutical compositions comprising a protein of the present invention and a pharmaceutically acceptable carrier are also provided. The present invention further provides for compositions comprising an antibody which specifically reacts with a protein of the present invention. Methods of identifying an inhibitor of IL-11 binding to the human IL-11 receptor are also provided. These methods comprise: (a) combining a human IL-11R protein or a fragment thereof with IL-11 or a fragment thereof, said combination forming a first binding mixture; (b) measuring the amount of binding between the protein and the IL- 11 or fragment in the first binding mixmre; (c) combining a compound with the protein and the IL-11 or fragment to form a second binding mixmre; (d) measuring the amount of binding in the second binding mixmre; and (e) comparing the amount of binding in the first binding mixture with the amount of binding in the second binding mixture; wherein the compound is capable of inhibiting IL-11 binding to the human IL- 11 receptor when a decrease in the amount of binding of the second binding mixture occurs. Optionally, the first and/or second binding mixture may further comprise gpl30 or a fragment thereof capable of binding to the protein of claim 11 or the IL-11 or fragment used therein. Inhibitors of IL- l lR identified by these methods and pharmaceutical compositions containing them are also provided. Methods of inhibiting binding of IL-11 to the human IL-1 1 receptor in a mammalian subject are also disclosed which comprise administering a therapeutically effective amount of a composition containing a human IL- l l R protein, an IL-llR inhibitor or an antibody to a human IL-l lR protein. Methods of treating or preventing loss of bone mass in a mammalian subject using these compositions are also provided. Brief Description of the Figures Figure 1 depicts a schematic representation of the structures of the human IL-11 receptor and gpl30. Figure 2 presents data demonstrating the biological activity of a soluble form of recombinant human IL-l lR protein. Detailed Description of Preferred Embodiments The inventors of the present application have for the first time identified and provided a polynucleotide encoding the human IL-11 receptor (human IL- l lR). SEQ ID NO: 1 provides the nucleotide sequence of a cDNA encoding the human IL-l lR. SEQ ID NO:2 provides the amino acid sequence of the receptor, included a putative signal sequence from amino acids 1-23. The mature human IL-l lR is believed to have the sequence of amino acids 24-422 of SEQ ID NO:2. The mature receptor has at least three distinct domains: an extracellular domain (comprising approximately amino acids 24-365 of SEQ ID NO: 2), a transmembrane domain (comprising approximately amino acids 366-390 of SEQ ID NO:2) and an intracellular domain (comprising approximately amino acids 391 -422 of SEQ ID NO:2). The extracellular domain is further divided into an immunoglobulin-like domain (comprising approximately amino acids 24-111 of SEQ ID NO: 2) and a type-I-cytokine domain (comprising approximately amino acids 112-365 of SEQ ID NO:2). Soluble forms of human IL-l lR protein can also be produced. Such soluble forms include without limitation proteins comprising amino acids 1-365 and 24-365 of SEQ ID NO:2. The soluble forms of the human IL-l lR are further characterized by being soluble in aqueous solution, preferably at room temperature. Human IL-11R proteins comprising only the intracellular domain or a portion thereof may also be produced. Any forms of human IL-l lR of less than full length are encompassed within the present invention and are referred to herein collectively as "human IL-llR" or "human IL-llR proteins. " Human IL-11R proteins of less than full length may be produced by expressing a corresponding fragment of the polynucleotide encoding the full-length human IL-11R protein (SEQ ID NO: 1). These corresponding polynucleotide fragments are also part of the present invention. Modified polynucleotides as described above may be made by standard molecular biology techniques, including construction of appropriate desired deletion mutants, site-directed mutagenesis methods or by the polymerase chain reaction using appropriate oligonucleόtide primers. Based upon similarity to the structure of the IL-6 receptor, it is predicted that IL-l lR proteins containing only the type-I cytokine domain of the extracellular region of the full length receptor will be capable of binding IL- 1 1 and inducing receptor signalling. As a result, IL-l lR proteins comprising amino acids 1 12 to 365 of SEQ ID NO:2, IL-l lR proteins comprising amino acids 112 to 390 of SEQ ID NO:2, and IL-11R proteins comprising amino acids 112 to 422 of SEQ ID NO:2 are provided by the present invention. Polynucleotides encoding such proteins (such as for example a polynucleotide comprising the nucleotides sequence of SEQ ID NO: l from nucleotide 1067 to nucleotide 1828, a polynucleotide comprising the nucleotides sequence of SEQ ID NO: l from nucleotide 1067 to nucleotide 1906, and a polynucleotide comprising the nucleotides sequence of SEQ ID NO: 1 from nucleotide 1067 to nucleotide 1999, respectively) are also provided by the invention. For the purposes of the present invention, a protein has "a biological activity of the human IL-11 receptor" if it possess one or more of the following characteristics: (1) the ability to bind IL-11 or a fragment thereof (preferably a biologically active fragment thereof); (2) the ability to bind to cytosolic proteins or molecules involved in the signalling pathway invoked by IL-11 binding to human IL-l lR; (3) the ability to produce a signal characteristic of the binding of IL-11 to human IL-l lR (where the protein in question either contains a portion able to bind IL-11 or where the protein in question would produce such signal if joined to another protein which is able to bind IL-11); (4) the ability to bind to gpl30 or a fragment thereof (either in the presence or absence of IL-11); (5) the ability to induce tyrosine phosphorylation of gpl30; (6) the ability to induce tyrosine phosphorylation of JAK kinases; or (7) the ability to induce tyrosine phosphorylation of the STAT family of DNA binding proteins (Zhong et al. (1994) Science 264, 95-98). Preferably, the biological activity possessed by the protein is the ability to bind IL- 1 1 or a fragment hereof, more prefereably with a K D of about 0.1 to about 100 nM, most preferably with a K D of about 1 to about 10 nM. Human IL-l lR or active fragments thereof (human IL-l lR proteins) may be fused to carrier molecules such as immunoglobulins. For example, soluble forms of the human IL-l lR may be fused through "linker" sequences to the Fc portion of an immunoglobulin. Other fusions proteins, such as those with GST, Lex-A or MBP, may also be used. The invention also encompasses allelic variations of the nucleotide sequence as set forth in SEQ ID NO: l . that is, naturally-occurring alternative forms of the isolated polynucleotide of SEQ ID NO: 1 which also encode human IL-l lR proteins, preferably those proteins having a biological activity of human IL-llR. Also included in the invention are isolated polynucleotides which hybridize to the nucleotide sequence set forth in SEQ ID NO: l under highly stringent conditions (for example, O. lxSSC at 65°C). Isolated polynucleotides which encode human IL-11R proteins but which differ from the nucleotide sequence set forth in SEQ ID NO: l by virtue of the degeneracy of the genetic code are also encompassed by the present invention. Variations in the nucleotide sequence as set foπh in SEQ ID NO: 1 which are caused by point mutations or by induced modifications are also included in the invention. The isolated polynucleotides of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al. , Nucleic Acids Res. 19. 4485-4490 ( 1991 ), in order to produce the human IL-11R protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing 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 ligated to form a covalent bond between the isolated polynucleotide of the invention and the expression control sequence, in such a way that the human IL-l lR protein is expressed by a host cell which has been transformed (transfected) with the ligated polynucleotide/expression control sequence. A number of types of cells may act as suitable host cells for expression of the human IL-l lR protein. Any cell type capable of expressing functional human IL-l lR protein may be used. Suitable mammalian host cells include, for example, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro culture of primary tissue, primary explants, HeLa cells, mouse L cells, BHK, HL-60, U937, HaK, Rat2, BaF3, 32D, FDCP-1 , PC12, Mix or C2C12 cells. The human IL-l lR protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences iri one or more insect expression vectors, and employing an insect expression system. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e.g. , Invitrogen, San Diego, California, U.S.A. (the MaxBac ® kit), and such methods are well known in the art. as described in Summers and Smith. Texas Agricultural Experiment Station Bulletin No. 1555 (1987), incorporated herein by reference. Soluble forms of the human IL-l lR protein may also be produced in insect cells using appropriate isolated polynucleotides as described above. Alternatively, the human IL-l lR protein may be produced in lower eukaryotes such as yeast or in prokaryotes such as bacteria. Suitable yeast strains include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins. Suitable bacterial strains include Escherichia coli, Bacillus subtilis, Salmonella typhimunum, or any bacterial strain capable of expressing heterologous proteins. Expression in bacteria may result in formation of inclusion bodies incorporating the recombinant protein. Thus, refolding of the recombinant protein may be required in order to produce active or more active material. Several methods for obtaining correctly folded heterologous proteins from bacterial inclusion bodies are known in the art. These methods generally involve solubilizing the protein from the inclusion bodies, then 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 accomplish the refolding in an environment which allows correct formation of disulfide bonds (a redox system). General methods of refolding are disclosed in Kohno, Meth. Enzvm.. 185: 187-195 (1990). EP 0433225 and copending application USSN 08/163,877 describe other appropriate methods. The human IL- l lR protein of the invention may also be expressed as a product of transgenic animals, e.g. , as a component of the milk of transgenic cows, goats, pigs, or sheep which are characterized by somatic or germ cells containing a polynucleotide sequence encoding the human IL-l lR protein. The human IL-11R protein of the invention may be prepared by growing a culture transformed host cells under culture conditions necessary to express the desired protein. The resulting expressed protein may then be purified from the culture medium or cell extracts. Soluble forms of the human IL-l lR protein of the invention can be purified from conditioned media. Membrane- bound forms of human IL-l lR protein of the invention can be purified by preparing a total membrane fraction from the expressing cell and extracting the membranes with a non-ionic detergent such as Triton X-100. The human IL-l lR protein can be purified using methods known to those skilled in the art. For example, the human IL-llR protein of the invention can be concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. Following the concentration step, the concentrate can be applied to a purification matrix such as a gel filtration medium. Alternatively, an anion exchange resin can be employed, for example, a matrix or substrate having pendant diethylaminoethyl (DEAE) or polyetheyleneimine (PEI) groups. ' The matrices can be acrylamide, agarose, dextran, cellulose or other types commonly employed in protein purification. Alternatively, a cation exchange step can be employed. Suitable cation exchangers include various insoluble matrices comprising sulfopropyl or carboxymethyl groups. Sulfopropyl groups are preferred (e.g. , S-Sepharose ® columns). The purification of the human IL- 1 1R protein from culture supernatant may also include one or more column steps over such affinity resins as concanavalin A-agarose, heparin-toyopearl ® or Cibacrom blue 3GA Sepharose ® ; or by hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether; or by immunoaffinity chromatography. Finally, one or more reverse-phase high performance liquid chromatography (RP-HPLC) steps employing hydrophobic RP-HPLC media, e.g. , silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the human IL-11R protein. Affinity columns including IL-11 or fragments thereof or including antibodies to the IL- l lR protein can also be used in purification in accordance with known methods. Some or all of the foregoing purification steps, in various combinations or with other known methods, can also be employed to provide a substantially purified isolated recombinant protein. Preferably, the isolated human IL-l lR protein is purified so that it is substantially free of other mammalian proteins. Human IL-llR proteins of the invention may also be used to screen for agents which are capable of binding to human IL-l lR or interfere with the binding of IL-11 to the human IL-l lR (either the extracellular or intracellular domains) and thus may act as inhibitors of normal binding and cytokine action (IL-l lR inhibitors). Binding assays using a desired binding protein, immobilized or not, are well known in the art and may be used for this purpose using the human IL-1 1R protein of the invention. Purified cell based or protein based (cell free) screening assays may be used to identify such agents. For example, human IL-l lR protein may be immobilized in purified form on a carrier and binding to purified human IL-l lR protein may be measured in the presence and in the absence of potential inhibiting agents. A suitable binding assay may alternatively employ a soluble form of human IL-l lR of the invention. In such a screening assay, a first binding mixture is formed by combining IL-11 or a fragment thereof and human IL-l lR protein, and the amount of binding in the first binding mixture (B 0 ) is measured. A second binding mixmre is also formed by combining IL-11 or a fragment thereof, human IL-l lR protein, and the compound or agent to be screened, and the amount of binding in the second binding mixmre (B) is measured. The amounts of binding in the first and second binding mixtures are compared, for example, by performing a calculation of the ratio B/B 0 . A compound or agent is considered to be capable of inhibiting binding if a decrease in binding in the second binding mixmre as compared to the first binding mixmre is observed. Optionally, gpl30 can be added to one or both of the binding mixtures. The formulation and optimization of binding mixtures is within the level of skill in the art, such binding mixtures may also contain buffers and salts necessary to enhance or to optimize binding, and additional control assays may be included in the screening assay of the invention. Compounds found to reduce the binding activity of human IL-l lR protein to IL-11 or its fragment to any degree, preferably by at least about 10% , more preferably greater than about 50% or more, may thus be identified and then secondarily screened in other binding assays and in vivo assays. By these means compounds having inhibitory activity for IL-l lR binding which may be suitable as therapeutic agents may be identified. Human IL-l lR proteins, and polynucleotides encoding them, may also be used as diagnostic agents for detecting the expression or presence of IL-11R, IL-11 or cells expressing IL-l lR or IL-11. The proteins or polynucleotides may be employed for such purpose in standard procedures for diagnostics assays using these types of materials. Suitable methods are well known to those skilled in the art. Human IL-l lR acts as a mediator of the known biological activities of IL-1 1. As a result, isolated human IL-11R protein and IL-1 1R inhibitors may be useful in treatment or modulation of various medical conditions in which IL- 11 is implicated or which are effected by the activity (or lack thereof) of IL-11 (collectively "IL-11 -related conditions"). IL-11-related conditions include without limitation immune deficiencies, specifically deficiencies in hematopoietic progenitor cells, or disorders relating thereto, cancer and other disease. Such pathological states may result from disease, exposure to radiation or drugs, and include, for example, leukopenia, bacterial and viral infections, anemia, B cell or T cell deficiencies such as immune cell or hematopoietic cell deficiency following a bone marrow transplantation. It is also believed that IL-1 1 and IL-l lR may play a role in the regulation of bone maturation and repair (Girasole et al. ( 1994) J. Clin. Invest. 93, 1516-1524; Passed et al. (1992) J. Bone Miner. Res.. 7(S1 ), SI 10 Abst. : Passeri et al. (1993) J. Bone Miner. Res. , 8(S1), S162 Abst. ). As a result, human IL-11R protein and IL-1 1R inhibitors may be useful in treatment of bone loss (including that associated with osteoporosis, post-menopausal osteoporosis. senile osteoporosis, idiopathic osteoporosis, Pagets disease, multipe myeloma, and hypogonadal conditions). Human IL-llR protein and IL-l lR inhibitors, purified from cells or recombinantly produced, may be used as a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Such a composition may contain, in addition to human IL-l lR or ligand and carrier, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials 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 route of administration. 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-15, G-CSF, stem cell factor, and erythropoietin. The pharmaceutical composition may contain thrombolytic or anti-thrombotic factors such as plasminogen activator and Factor VIII. The pharmaceutical composition may further contain other anti-inflammatory agents. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with isolated human IL-l lR protein or IL-l lR inhibitor, or to minimize side effects caused by the isolated human IL-l lR or IL- l lR inhibitor. Conversely, isolated human IL-l lR or IL-l lR inhibitor may 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, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti- inflammatory agent. The pharmaceutical composition of the invention may be in the form of a liposome in which isolated human IL-l lR protein or IL-l lR inhibitor is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers which in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Patent No. 4,235,871; U.S. Patent No. 4,501,728; U.S. Patent No. 4,837,028; and U.S. Patent No. 4,737,323, all of which are 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 meaningful patient benefit, e.g. , amelioration of symptoms of, healing of, or increase in rate of healing of such conditions. When applied to an individual active ingredient, 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 practicing the method of treatment or use of the present invention, a therapeutically effective amount of isolated human IL-l lR protein or IL- l lR inhibitor is administered to a mammal. Isolated human IL-l lR protein or IL- l lR inhibitor may be administered in accordance with 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, human IL-llR protein or IL-l lR inhibitor may be administered either simultaneously with the cytokine(s). lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the attending physician will decide on the appropriate sequence of administering human IL-l lR protein or IL-l lR inhibitor in combination with cytokine(s), lymphokine(s), other hematopoietic factor(s). thrombolytic or anti-thrombotic factors. Administration of human IL-11R protein or IL-11R inhibitor used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion. inhalation, or cutaneous, subcutaneous, or intravenous injection. Intravenous administration to the patient is preferred. When a therapeutically effective amount of human IL-1 1R protein σr IL- 11R inhibitor is administered orally, human IL-l lR protein or IL-1 1R inhibitor 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 a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95% human IL- 1 1 R protein or IL-l lR inhibitor, and preferably from about 25 to 90% human IL- l l R protein or IL-llR inhibitor. When administered in liquid form, a liquid carrier such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline solution, 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 human IL-l lR protein or IL-l lR inhibitor, and preferably from about 1 to 50% human IL-l lR protein or IL-l lR inhibitor. When a therapeutically effective amount of human IL- 11R protein or IL- 11R inhibitor is administered by intravenous, cutaneous or subcutaneous injection, human IL-llR protein or IL-llR inhibitor will be in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable protein solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art. A preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection should contain, in addition to human IL-l lR protein or IL-l lR inhibitor an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection. Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringer's Injection, or other vehicle as known in the an. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additive known to those of skill in the art. The amount of human IL- l lR protein or IL- l lR inhibitor in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone. Ultimately, the attending physician will decide the amount of human IL-11R protein or IL-11R inhibitor with which to treat each individual patient. Initially, the attending physician will administer low doses of human IL-l lR protein or IL-l lR inhibitor and observe the patient's response. Larger doses of human IL-11R protein or IL-11R inhibitor may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not generally increased further. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.1 μg to about 100 mg of human IL-l lR protein or IL-llR inhibitor per kg 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 condition and potential idiosyncratic response of each individual patient. It is contemplated that the duration of each application of the human IL-l lR protein or IL-l lR inhibitor will be in the range of 12 to 24 hours of continuous intravenous administration. Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention. Human IL-l lR proteins of the invention may also be used to immunize animals to obtain polyclonal and monoclonal antibodies which specifically react with the human IL-l lR protein and which may inhibit binding of IL-1 1 or fragments thereof to the receptor. Such antibodies may be obtained using the entire human IL-l lR as an immunogen, or by using fragments of human IL- 11R, such as the soluble mature human IL-l lR. Smaller fragments of the human IL-l lR may also be used to immunize animals. The peptide immunogens additionally may contain a cysteine residue at the carboxyl terminus, and are conjugated to a hapten such as keyhole limpet hemocyanin (KLH). Additional peptide immunogens may be generated by replacing tyrosine residues with sulfated tyrosine 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). Neutralizing or non-neutralizing antibodies (preferably monoclonal antibodies) binding to human IL-llR protein may also be useful therapeutics for certain tumors and also in the treatment of conditions described above. These neutralizing monoclonal antibodies may be capable of blocking IL-11 binding to the human IL-llR. Example 1 Isolation of Human IL-l lR cDNA Generation of DNA Probes: DNA probes derived from the murine Etl-2 sequence (SEQ ID NO: 3) were obtained by PCR from murine placenta cDNA. The amino terminal probe corresponds to base pairs 418-570 and the carboxy terminal probe to base pairs 847- 1038 of the murine Etl-2 sequence. The DNA probes were gel purified and radiolabeled using 32P-dATP and α32P-dCTP. ι-ι cDNA Library Screening: cDNA was generated from activated human PBMC using the Superscript Choice System and cloned into the EcoRl site of ZAP II (Stratagene) . The resulting phage were used to infect E. coli strain BB4. One million phage were plated on 150 mm NZCYM plates at a density of 15000 pfu/plate. Plaques were transferred to duplicate Duralose nitrocellulose filters (Stratagene). Following alkali denaturation and heat fixation the filters were pre-hybridized in 5X SSC, 5X Denhardts, 0.1 % SDS, and 50 μg/ml yeast tRNA for 2 hours at 65°C. One set of filters was hybridized with the amino-terminal probe and the other set with the carboxy-terminal probe (5 x 10 5 cpm/ml) for 48 hrs at 55°C in pre-hybridization buffer. The filters were washed with 4X SSC, 0.1 % SDS once at 25°C and twice at 55°C. Plaques that hybridized to both probes were identified by autoradiography. Of the one million plaques screened two plaques hybridized to both of the probes. These plaques were picked and the phage eluted into SM media containing chloroform. The resulting phage were used to reinfect E. coli strain BB4 and plated on NZCYM plates at a density of 100-300 pfu/plate for a secondary screen. Following the secondary screen plasmid DNA was isolated from the ZAPII plaques by excision using helper phage (Stratagene). The DNA sequence of the inserts was determined on an Applied Biosystems DNA sequencer. Clone phILHR14-2 containing the polynucleotide having the sequence of SEQ ID NO: 1 was deposited with ATCC at accession number on December 22, 1994. Example 2 Expression of Soluble Human IL-l lR Protein and Assay of Activity A soluble form of human IL-l lR protein was expressed in mammalian cells. The expresssed recombinant protein was capable of transducing a signal in BAF 130-9 cells. A portion of the full length human IL-l lR sequence (nucleotides 734- 1828 of SEQ ID NO: l encoding amino acids 1-365 of SEQ ID NO:2) corresponding to a soluble form was cloned into the mammalian expression vector pED and used to transfect COSM6 cells. 40 hours after transfection conditioned media was removed, concentrated 5 fold and used in proliferation assays with the murine cell line BAF130-9 (Hibi, M. et al. (1990) Cell 63, 1 149-57), a derivative of the BAFB03 cell line expressing the human gpl30 signal transducer. BAF 130-9 cells do not proliferate in response to IL-1,1 or IL-6 alone, but do proliferate in response to a combination of IL-6 and soluble IL-6R (Hibi et al. , supra). BAF130-9 cells (lxlO 4 cell in 0.1 ml) were cultured in RPMI 1640 medium/ 10% FCS with increasing concentrations of recombinant human IL- 1 1 in the absence or presence of 10 μ\ of conditioned media from mock transfected cells or cells transfected with the soluble human IL- l lR sequence. After forty hours the cells were pulse-labeled with 3 H-thymidine (0.5 μCi/well) for eight hours and incorporated nucleotide was determined. As shown in Figure 2, BAF130-9 cells do not proliferate in response to IL-11 or soluble IL-l lR alone, but do proliferate in the presence of both IL-11 and soluble IL-l lR. Other human IL-l lR proteins can be tested in this model to determine whether they exhibit a "biological activity" of human IL-11R as defined herein. Example 3 Other Systems for Determination Biological Activity of Human IL-11R Protein Other systems can be used to determine whether a specific human IL- 11R protein exhibits a "biological activity" of human IL-11R as defined herein. The following are examples of such systems. Assays for IL-11 Binding The ability of a human IL-l lR protein to bind IL-11 or a fragment thereof can be determine by any sutiable assays which can detect such binding. Some suitable examples follow. Binding of IL-11 to the extracellular region of the human IL-11R protein will specifically cause a rapid induction of phosphotyrosine on the receptor protein. Assays for ligand binding activity as measured by induction of phosphorylation are described below. Alternatively, a human IL-l lR protein (such as, for example, a soluble form of the extracellular domain) is produced and used to detect IL- 1 1 binding. For example, a DNA construct is prepared in which the extracellular domain (truncated prior, preferably immediately prior, to the predicted transmembrane domain) is ligated in frame to a cDNA encoding the hinge C H 2 and C H 3 domains of a human immunoglobulin (Ig) γl. This construct is generated in an appropriate expression vector for COS cells, such as pEDΔC or pMT2. The plasmid is transiently transfected into COS cells. The secreted IL-1 lR-Ig fusion protein is collected in the conditioned medium and purified by protein A chromatography. The purified IL-l lR-Ig fusion protein is used to demonstrate IL-11 binding in a number of applications. IL-11 can be coated onto the surface of an enzyme-linked immunosorbent assay (ELISA) plate, and then additional binding sites blocked with bovine serum albumin or casein using standard ELISA buffers. The IL-llR-Ig fusion protein is then bound to the solid-phase IL-11 , and binding is detected with a secondary goat anti-human Ig conjugated to horseradish peroxidase. The activity of specifically bound enzyme can be measured with a colorimetric substrate, such as tetramethyl benzidine and absorbance readings. IL-1 1 may also be expressed on the surface of cells, for example by providing a transmembrane domain or glucosyl phosphatidyl inositol (GPI) linkage. Cells expressing the membrane bound IL-11 can be identified using the IL-l lR-Ig fusion protein. The soluble IL-l lR-Ig fusion is bound to the surface of these cells and detected with goat anti-human Ig conjugated to a fluorochrome, such as fluorescein isothiocyanate and flow cytometrv. Interaction Trap A yeast genetic selection method, the "interaction trap" [Gyuris et al, Cell 75:791-803, 1993], can be used to determine whether a human IL-l lR protein has a biological activity of human IL-l lR as defined herein. In this system, the expression of reporter genes from both LexAop-Leu2 and LexAop-LacZ relies on the interaction between the bait protein, for example in this case a species which interacts with huamn IL-l lR, and the prey, for example in this case the human IL-l lR protein. Thus, one can measure the strength of the interaction by the level of Leu2 or LacZ expression. The most simple method is to measure the activity of the LacZ encoded protein. /3-galactosidase. This activity can be judged by the degree of blueness on the X-Gal containing medium or filter. For the quantitative measurement of 0-galactosidase activity, standard assays can be found in "Methods in Yeast Genetics" Cold Spring Harbor, New York, 1990 (by Rose, M.D. , Winston, F. , and Hieter, P.). In such methods, if one wishes to determine whether the human IL-1 1R protein interacts with a particular species (such as, for example, a cystoslic protein which binds to the intracellular domain of the human IL-l lR in vivo). that species can be used as the "bait" in the interaction trap with the human 1L- 11R protein to be tested serving as the "prey", or vice versa. CAT Induction System Transcription of acute phase plasma protein genes, such as the rat /3-fibrinogen gene, is activated by IL-1 1 in a varietj of cell lines. In one exemplary system, COSM6 cells are cotransfected with plasmids encoding the human IL-11R protein (such as the full length human IL-11R or a soluble form thereof), the human gpl30 signal transducer and a reporter gene containing the 350 base pair promoter region of the rat b-fibrinogen gene fused to a reporter gene, CAT (Baumann et al. (1991) J. Biol. Chem. 266, 20424-27). The cells are stimulated with increasing concentrations of recombinant human IL-11 and transcription of the reporter gene is monitored by assaying for the presence of CAT activity. Phosphorylation of gp!30 Activity may also be determined by examining the ability of IL-11 to induce tyrosine phosphorylation of gpl30 in cells transfected with a sequence encoding the human IL-l lR protein (such as the full length human IL-llR or a soluble form thereof) (Luttcken et al. (1994) Science 263, 89-92). Phosphorylation of STATs Activity may also be determined by examining the ability of IL-1 1 to induce tyrosine phosphorylation of STATs (signal transducers and activators of transcription, a family of DNA binding proteins) in cells transfected with a sequence encoding the human IL-11R protein (such as the full length human IL- 11R or a soluble form thereof) (Zhong et al. (1994) Science 264. 95-98) Phosphorylation of JAK Kinases Activity may also be determined by examining the ability of IL-11 to induce tyrosine phosphorylation of JAK kinases in cells transfected with a sequence encoding die human IL-1 IR protein (such as the full length human IL- 11R or a soluble form thereof) (Yin et al. (1993) J. Immunol. 151 : 2555-61). All patent and literature references cited herein are incorporated by reference as if fully set forth. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Tobin, James (ii) TITLE OF INVENTION: HUMAN INTERLUEKIN-11 RECEPTOR (iii) NUMBER OF SEQUENCES: 4 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Genetics Institute, Inc. (B) STREET: 87 CambridgePark Drive (C) CITY: Cambridge (D) STATE: MA (E) COUNTRY: USA (F) ZIP: 02140 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Brown, Scott A. (B) REGISTRATION NUMBER: 32,724 (C) REFERENCE/DOCKET NUMBER: GI5252 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (617) 498-8224 (B) TELEFAX: (617) 876-5851 ( 2 ) INFORMATION FOR SEQ ID NO: 1 : (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2456 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (il) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 734..1999 (κi) SEQUENCE DESCRIPTION: SEQ ID NO:1: TCGCCCACCC CCAGCCTCTG GCAGCAGCCA GGGCATCTGG ATCTGCTTAA CTACACAGCC 60 J CCAGCCTGCA CCCTAGCCCC ATCCAGCTTC ACAAACTGGA GACCAACGAA GTGTCAAGAG 120 CCAGGCCCAG CTGAGTGGCC CAAGTAGCCA GACCAAGGAG CCAGGTTCAG GCGAGAAGCC 180 TGGCAGCCAG GGCAGGGGTG GGCCTCAGGG TGGGAGTGCA GGATGGGCTC AGATCCATGA 240 TGACACCCTT CCCCCAGGGT GATAAGGTCT GCCTAGGTTA ATCAGAGGCA GTGATAAGCC 300 CTGGACCAGG TGGGGGTAAA TACCAGAATT CCCAACAGCT GGACTGGAGG GGTTAATGGG 360 AGTGGCTGAG CTGGTGCCAG TGCTTGGTGC CAGGGGTGGG CGCCAAGGGC AGTGGAGGGG 420 GAGTTGCTGG CACAGTCTGT TGCCTCCGGC TTTTGTTCTG GGCCCTAAGC CCAGGACTGA 480 GATGGAGGGT GTGAGGGGGT GTGTGTGTCC GTGTGTGTGT GTGTGTGTGT GTGCGCGCGC 540 ACGCACATGC AAAGCACTGG GTATACAGTG GGAAAGGGGA CCTCAGGTCA GTTCCCGCAG 600 TGATTTCTAA CAGCCTTACC CCACTTGGTG CATCAATTTT TCTCCTAGGA AGCCTCAGTT 660 TTGGAGAGGA AGAGCCAGGC TTTAGCCTCC CATCTCAGGG GTCGGGGATT TTTGACTCTA 720 769 GTG GCC GTG GCT ACA GCC CTG GTG TCT GCC TCC TCC CCC TGC CCC CAG 817 Val Ala Val Ala Thr Ala Leu Val Ser Ala Ser Ser Pro Cys Pro Gin 15 20 25 GCC TGG GGC CCC CCA GGG GTC CAG TAT GGG CAG CCA GGC AGG TCC GTG 865 Ala Trp Gly Pro Pro Gly Val Gin Tyr Gly Gin Pro Gly Arg Ser Val 30 35 40 AAG CTG TGT TGT CCT GGA GTG ACT GCC GGG GAC CCA GTG TCC TGG TTT 913 Lys Leu Cys Cys Pro Gly Val Thr Ala Gly Asp Pro Val Ser Trp Phe 45 50 55 60 CGG GAT GGG GAG CCA AAG CTG CTC CAG GGA CCT GAC TCT GGG CTA GGG 961 Arg Asp Gly Glu Pro Lys Leu Leu Gin Gly Pro Asp Ser Gly Leu Gly 65 70 75 l\J CAT GAA CTG GTC CTG GCC CAG GCA GAC AGC ACT GAT GAG GGC ACC TAC 1009 His Glu Leu Val Leu Ala Gin Ala Asp Ser Thr Asp Glu Gly Thr Tyr 80 85 90 ATC TGC CAG ACC CTG GAT GGT GCA CTT GGG GGC ACA GTG ACC CTG CAG 1057 He Cys Gin Thr Leu Asp Gly Ala Leu Gly Gly Thr Val Thr Leu Gin 95 100 105 CTG GGC TAC CCT CCA GCC CGC CCT GTT GTC TCC TGC CAA GCA GCC GAC 1105 Leu Gly Tyr Pro Pro Ala Arg Pro Val Val Ser Cys Gin Ala Ala Asp 110 115 120 TAT GAG AAC TTC TCT TGC ACT TGG AGT CCC AGC CAG ATC AGC GGT TTA 1153 Tyr Glu Asn Phe Ser Cys Thr Trp Ser Pro Ser Gin lie Ser Gly Leu : P:r:o «Thr A «rg ~Tyr „Leu :Th:r ra Ser -Ty.r A ~rg. *L,ys :Lyιs »Thr V -a.l «Leu „Gly A ::la 145 150 155 ~- GAT AGC CAG AGG AGG AGT CCA TCC ACA GGG CCC TGG CCA TGC CCA CAG 1249 Asp Ser Gin Arg Arg Ser Pro Ser Thr Gly Pro Trp Pro Cys Pro Gin 160 165 170 GAT CCC CTA GGG GCT GCC CGC TGT GTT GTC CAC GGG GCT GAG TTC TGG 1297 Asp Pro Leu Gly Ala Ala Arg Cys Val Val His Gly Ala Glu Phe Trp 175 180 185 AGC CAG TAC CGG ATT AAT GTG ACT GAG GTG AAC CCA CTG GGT GCC AGC 1345 Ser Gin Tyr Arg lie Asn Val Thr Glu Val Asn Pro Leu Gly Ala Ser 190 195 200 ACA CGC CTG CTG GAT GTG AGC TTG CAG AGC ATC TTG CGC CCT GAC CCA 1393 Thr Arg Leu Leu Asp Val Ser Leu Gin Ser lie Leu Arg Pro Asp Pro 205 210 215 220 CCC CAG GGC CTG CGG GTA GAG TCA GTA CCA GGT TAC CCC CGA CGC CTG 1441 Pro Gin Gly Leu Arg Val Glu Ser Val Pro Gly Tyr Pro Arg Arg Leu 225 230 235 CO CGA GCC AGC TGG ACA TAC CCT GCC TCC TGG CCG TGC CAG CCC CAC TTC 1489 w Arg Ala Ser Trp Thr Tyr Pro Ala Ser Trp Pro Cys Gin Pro His Phe 240 245 250 CTG CTC AAG TTC CGT TTG CAG TAC CGT CCG GCG CAG CAT CCA GCC TGG 1537 Leu Leu Lys Phe Arg Leu Gin Tyr Arg Pro Ala Gin His Pro Ala Trp 255 260 265 TCC ACG GTG GAG CCA GCT GGA CTG GAG GAG GTG ATC ACA GAT GCT GTG 1585 Ser Thr Val Glu Pro Ala Gly Leu Glu Glu Val He Thr Asp Ala Val 270 275 280 GCT GGG CTG CCC CAT GCT GTA CGA GTC AGT GCC CGG GAC TTT CTA GAT 1633 Ala Gly Leu Pro His Ala Val Arg Val Ser Ala Arg Asp Phe Leu Asp 285 290 295 300 GCT GGC ACC TGG AGC ACC TGG AGC CCG GAG GCC TGG GGA ACT CCG AGC 1681 Ala Gly Thr Trp Ser Thr Trp Ser Pro Glu Ala Trp Gly Thr Pro Ser 305 310 315 ACT GGG ACC ATA CCA AAG GAG ATA CCA GCA TGG GGC CAG CTA CAC ACG 1729 Thr Gly Thr He Pro Lys Glu He Pro Ala Trp Gly Gin Leu His Thr 320 325 330 CAG CCA GAG GTG GAG CCT CAG GTG GAC AGC CCT GCT CCT CCA AGG CCC 1777 Gin Pro Glu Val Glu Pro Gin Val Asp Ser Pro Ala Pro Pro Arg Pro 335 340 345 TCC CTC CAA CCA CAC CCT CGG CTA CTT GAT CAC AGG GAC TCT GTG GAG 1825 Ser Leu Gin Pro His Pro Arg Leu Leu Asp His Arg Asp Ser Val Glu 350 355 360 CAG GTA GCT GTG CTG GCG TCT TTG GGA ATC CTT TCT TTC CTG GGA CTG 1873 Gin Val Ala Val Leu Ala Ser Leu Gly He Leu Ser Phe Leu Gly Leu 365 370 375 380 GTG GCT GGG GCC CTG GCA CTG GGG CTC TGG CTG AGG CTG AGA CGG GGT 1921 Val Ala Gly Ala Leu Ala Leu Gly Leu Trp Leu Arg Leu Arg Arg Gly 385 390 395 GGG AAG GAT GGA TCC CCA AAG CCT GGG TTC TTG GCC TCA GTG ATT CCA 1969 Gly Lys Asp Gly Ser Pro Lys Pro Gly Phe Leu Ala Ser Val lie Pro 400 405 410 ° J GTG GAC AGG CGT CCA GGA GCT CCA AAC CTG TAGAGGACCC AGGAGGGCTT 2019 Val Asp Arg Arg Pro Gly Ala Pro Asn Leu 415 420 CGGCAGATTC CACCTATAAT TCTGTCTTGC TGGTGTGGAT GGATGGACAG ATAGAAACCA 2079 GGCAGGACAG TAGATCCCTA TGGTTGGATC TCAGCTGGAA GTTCTGTTTG GAGCCCATTT 2139 CTGTGAGACC CTGTATTTCA AATTTGCAGC TGAAAGGTGC TTGTACCTCT GATTTCACCC 2199 CAGAGTTGGA GTTCTGCTCA AGGAACGTGT GTAATGTGTA CATCTGTGTC CATGTGTGAC 2259 CATGTGTCTG TGAGGCAGGG AACATGTATT CTCTGCATGC ATGTATGTAG GTGCCTGGGG 2319 AGTGTGTGTG GGTCCTTGGC TCTTGGCCTT TCCCCTTGCA GGGGTTGTGC AGGTGTGAAT 2379 AAAGAGAATA AGGAAGTTCT TGGAGATTAT ACTCAGAAAA AAAAAAAAAA AGTCGACGCG 2439 GCCGCGAATT CCTGCAG 2456 (2) INFORMATION FOR SEQ ID NO:2 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 422 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2 : Met Ser Ser Ser Cys Ser Gly Leu Ser Arg Val Leu Val Ala Val Ala 1 5 10 15 Thr Ala Leu Val Ser Ala Ser Ser Pro Cys Pro Gin Ala Trp Gly Pro 20 25 30 Pro Gly Val Gin Tyr Gly Gin Pro Gly Arg Ser Val Lys Leu Cys Cys 35 40 45 cυ Pro Gly Val Thr Ala Gly Asp Pro Val Ser Trp Phe Arg Asp Gly Glu * ■ " 50 55 60 Pro Lys Leu Leu Gin Gly Pro Asp Ser Gly Leu Gly His Glu Leu Val 65 70 75 80 Leu Ala Gin Ala Asp Ser Thr Asp Glu Gly Thr Tyr lie Cys Gin Thr 85 90 95 Leu Asp Gly Ala Leu Gly Gly Thr Val Thr Leu Gin Leu Gly Tyr Pro 100 105 110 Pro Ala Arg Pro Val Val Ser Cys Gin Ala Ala Asp Tyr Glu Asn Phe 115 120 125 Ser Cys Thr Trp Ser Pro Ser Gin He Ser Gly Leu Pro Thr Arg Tyr 130 135 140 Leu Thr Ser Tyr Arg Lys Lys Thr Val Leu Gly Ala Asp Ser Gin Arg 145 150 155 160 Pio Lys Glu He Pro Ala Trp Gly Gin Leu His Thr Gin Pro Glu Val 325 330 335 Glu Pro πln Val Asp Ser Pro Ala Pro Pro Arg Pro Ser Leu Gin Pro His Pro Leu Ala Ser Leu Gly He Leu Ser Phe Leu Gly Leu Val Ala Gly Ala 370 375 380 Leu Ala Leu Gly Leu Trp Leu Arg Leu Arg Arg Gly Gly Lys Asp Gly 385 390 395 400 Ser Pro Lys Pro Gly Phe Leu Ala Ser Val He Pro Val Asp Arg Arg 405 410 415 Pro Gly Ala Pro Asn Leu 420 (2) INFORMATION FOR SEQ ID NO: 3 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1714 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA ω *»J (iii) HYPOTHETICAL: NO (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 3 ..1359 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3 : TTCTTAGCCT GATAGGAGGA AGTCTTGGAG GCC ATG GCA CTC AGT CAC TGT GAT 54 Met Ala Leu Ser His Cys Asp 1 5 TAT CAA GAT GAG CAG CAG CTG CTC AGG GCT GAC CAG GGT CCT GGT GGC 102 Tyr Gin Asp Glu Gin Gin Leu Leu Arg Ala Asp Gin Gly Pro Gly Gly 10 15 20 CGT GCT ACA GCC CTG GTG TCT TCC TCC TCC CCC TGC CCC CAA GCT TGG 150 Λi <j Λla Tin Ala Lou Val Ser Set S^i Ser Pro Cys Pro Gin Ala Trp 25 30 35 GGT CCT CCA GGG GTC CAG TAT GGA CAA CCT GGC AGG CCC GTG ATG CTG 198 Gly Pro Pro Gly Val Gin Tyr Gly Gin Pro Gly Arg Pro Val Met Leu 40 45 50 55 TGC TGC CCC GGA GTG AGT GCT GGG ACT CCA GTG TCC TGG TTT CGG GAT 246 Cys Cys Pro Gly Val Ser Ala Gly Thr Pro Val Ser Trp Phe Arg Asp 60 65 70 GGA GAT TCA AGG CTG CTC CAG GGA CCT GAC TCT GGG TTA GGA CAC AGA 294 Gly As Ser Arg Leu Leu Gin Gly Pro Asp Ser Gly Leu Gly His Arg 75 80 85 CTG GTC TTG GCC CAG GTG GAC AGC CCT GAT GAA GGC ACT TAT GTC TGC 342 Leu Val Leu Ala Gin Val Asp Ser Pro Asp Glu Gly Thr Tyr Val Cys 90 95 100 CAG ACC CTG GAT GGT GTA TCA GGG GGC ATG GTG ACC CTG AAG CTG GGC 390 Gin Thr Leu Asp Gly Val Ser Gly Gly Met Val Thr Leu Lys Leu Gly 105 110 115 O 00 TTT CCC CCA GCA CGT CCT GAA GTC TCC TGC CAA GCG GTA GAC TAT GAA 438 Phe Pro Pro Ala Arg Pro Glu Val Ser Cys Gin Ala Val Asp Tyr Glu 120 125 130 135 AAC TTC TCC TGT ACT TGG AGT CCA GGC CAG GTC AGC GGT TTG CCC ACC 486 Asn Phe Ser Cys Thr Trp Ser Pro Gly Gin Val Ser Gly Leu Pro Thr 140 145 150 CGC TAC CTT ACT TCC TAC AGG AAG AAG ACG CTG CCA GGA GCT GAG AGT 534 Arg Tyr Leu Thr Ser Tyr Arg Lys Lys Thr Leu Pro Gly Ala Glu Ser 155 160 165 CAG AGG GAA AGT CCA TCC ACC GGG CCT TGG CCG TGT CCA CAG GAC CCT 582 Gin Arg Glu Ser Pro Ser Thr Gly Pro Trp Pro Cys Pro Gin Asp Pro ™ Leu G G ft l G u :A:la ~ Se.r A _r»g „Cys V ™ al :V;a:l H „is „Gly „Ala „Glu :P;h:e „Trp _Se„r «Gl.u „. 185 190 195 TAC CGG ATC AAT GTG ACC GAG GTG AAC CCA CTG GGT GCC AGC ACG TGC 678 Tyr Arg He Asn Val Thr Glu Val Asn Pro Leu Gly Ala Ser Thr Cys 200 205 210 215 CTA CTG GAT GTG AGA TTA CAG AGC ATC TTG CGT CCT GAT CCA CCC CAA 726 Leu Leu Asp Val Arg Leu Gin Ser He Leu Arg Pro Asp Pro Pro Gin 220 225 230 GGA CTG CGG GTG GAA TCC GTA CCT GGT TAC CCG AGA CGC CTG CAT GCC 774 Gly Leu Arg Val Glu Ser Val Pro Gly Tyr Pro Arg Arg Leu His Ala 235 240 245 AGC TGG ACA TAC CCT GCC TCC TGG CGT CGC CAA CCC CAC TTT CTG CTC 822 Ser Trp Thr Tyr Pro Ala Ser Trp Arg Arg Gin Pro His Phe Leu Leu 250 255 260 AAG TTC CGG TTG CAA TAC CGA CCA GCA CAG CAT CCA GCC TGG TCC ACG 870 Lys Phe Arg Leu Gin Tyr Arg Pro Ala Gin His Pro Ala Trp Ser Thr 265 270 275 GTG GAG CCC ATT GGC TTG GAG GAA GTG ATA ACA GAT GCT GTG GCT GGG 918 {£ Val Glu Pro He Gly Leu Glu Glu Val He Thr Asp Ala Val Ala Gly 280 285 290 295 CTG CCA CAC GCG GTA CGA GTC AGT GCC AGG GAC TTT CTG GAT GCT GGC 966 Leu Pro His Ala Val Arg Val Ser Ala Arg Asp Phe Leu Asp Ala Gly 300 305 310 ACC TGG AGC GCC TGG AGC CCA GAG GCC TGG GGT ACT CCT AGC ACT GGT 1014 Thi Trp Ser Ala Trp Ser Pro Glu Ala Trp Gly Thr Pro Ser Thr Gly 315 320 325 CCC CTG CAG GAT GAG ATA CCT GAT TGG AGC CAG GGA CAT GGA CAG CAG 1062 Pro Leu Gin Asp Glu He Pro Asp Trp Ser Gin Gly His Gly Gin Gin 330 335 340 CTA GAG GCA GTA GTA GCT CAG GAG GAC AGC CCG GCT CCT GCA AGG CCT 1110 Leu Glu Ala Val Val Ala Gin Glu Asp Ser Pro Ala Pro Ala Arg Pro 345 350 355 TCC TTG CAG CCG GAC CCA AGG CCA CTT GAT CAC AGG GAC CCC TTG GAG 1158 Ser Leu Gin Pro Asp Pro Arg Pro Leu Asp His Arg Asp Pro Leu Glu 360 365 370 375 CAA GTA GCT GTG TTA GCG TCT CTG GGA ATC TTC TCT TGC CTT GGC CTG 1206 Gin Val Ala Val Leu Ala Ser Leu Gly He Phe Ser Cys Leu Gly Leu 380 385 390 GCT GTT GGA GCT CTG GCA CTG GGG CTC TGG CTG AGG CTG AGA CGG AGT 1254 Ala Val Gly Ala Leu Ala Leu Gly Leu Trp Leu Arg Leu Arg Arg Ser 395 400 405 GGG AAG GAT GGA CCG CAA AAA CCT GGG CTC TTG GCA CCC ATG ATC CCG 1302 Gly Lys Asp Gly Pro Gin Lys Pro Gly Leu Leu Ala Pro Met He Pro 410 415 420 GTG GAA AAG CTT CCA GGA ATT CCA AAC CTG CAG AGG ACC CCA GAG AAC 1350 Val Glu Lys Leu Pro Gly He Pro Asn Leu Gin Arg Thr Pro Glu Asn 425 430 435 TTC AGC TGATTTCATC TGTAACCCGG TCAGACTTGG GGTGGTTAAA AGGACAGGCA 1406 Phe Ser 440 GAAAGAGGCG GGGCAGTGGA TCCCTGTGGA TGGAGGTCTC AGCTGAAAGT CTGAGCTCTT 1466 TTCTTTGACA CCTATACTCC AAACTTGCTG CCGGCTGAAG GCTGTCTGGA CTTCCGATGT 1526 CCTGAGGTGG AAGTCCACCT GAGGAATGTG TACAGAAGTC TGTGTTCCTG TGATCGTGTG 1586 TGTATGTGAG ACAGGGAGCA AAAGTTCTCT GCATGTGTGT ACAGATGATT GGAGAGTGTG 1646 TGCGGTCTTG GGCTTGGCCC TTCTGGGAAG TGTGAAGAGT TGAAATAAAA GAGACGGAAG 1706 TTTTTGGA 1714 (2) INFORMATION FOR SEQ ID NO:4 : i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 441 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Met Ala Leu Ser His Cys Asp Tyr Gin Asp Glu Gin Gin Leu Leu Arg 1 5 10 15 Ala Asp Gin Gly Pro Gly Gly Arg Ala Thr Ala Leu Val Ser Ser Ser 20 25 30 Ser Pro Cys Pro Gin Ala Trp Gly Pro Pro Gly Val Gin Tyr Gly Gin 35 40 45 Pro Gly Arg Pro Val Met Leu Cys Cys Pro Gly Val Ser Ala Gly Thr 50 55 60 Pro Val Ser Trp Phe Arg Asp Gly Asp Ser Arg Leu Leu Gin Gly Pro 65 70 75 80 Asp Ser Gly Leu Gly His Arg Leu Val Leu Ala Gin Val Asp Ser Pro 85 90 95 Asp Glu Gly Thr Tyr Val Cys Gin Thr Leu Asp Gly Val Ser Gly Gly 100 105 110 Met Val Thr Leu Lys Leu Gly Phe Pro Pro Ala Arg Pro Glu Val Ser 115 120 125 Cys Gin Ala Val Asp Tyr Glu Asn Phe Ser Cys Thr Trp Ser Pro Gly 130 135 140 Gin Val Ser Gly Leu Pro Thr Arg Tyr Leu Thr Ser Tyr Arg Lys Lys 145 150 155 160 Thr Leu Pro Gly Ala Glu Ser Gin Arg Glu Ser Pro Ser Thr Gly Pro 165 170 175 Trp Pro Cys Pro Gin Asp Pro Leu Glu Ala Ser Arg Cys Val Val His 380 185 190 Gly Ala Glu Phe Trp Ser Glu Tyr Arg He Asn Val Thr Glu Val Asn 195 200 205 Cys Leu Leu Asp Val Arg Leu Gin Ser He Gin Gly Leu Arg Val Glu Ser Val Pro Gly 235 240 Ala Ser Trp Thr Tyr Pro Ala Ser Trp Arg 250 255 Leu Lys Phe Arg Leu Gin Tyr Arg Pro Ala 265 270 Thr Val Glu Pro He Gly Leu Glu Glu Val 280 285 Gly Leu Pro His Ala Val Arg Val Ser Ala 295 300 Gly Thr Trp Ser Ala Trp Ser Pro Glu Ala 315 320 ro Gly Pro Leu Gin Asp Glu He Pro Asp Trp 330 335 Gin Leu Glu Ala Val Val Ala Gin Glu Asp 345 350 Pro Ser Leu Gin Pro Asp Pro Arg Pro Leu 360 365 Glu Gin Val Ala Val Leu Ala Ser Leu Gly 375 380 Leu Ala Val Gly Ala Leu Ala Leu Gly Leu 395 400 Ser Gly Lys Pro V l Glu Lys Leu Pro Gly He Pro Asn 425 430 Leu Gin Arg Thr Pro Glu Asn Phe Ser 435 440 4-. O

Claims: What is claimed is: 1. An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: (a) the nucleotide sequence of SEQ ID NO: l from nucleotide 803 to nucleotide 1999; (b) a nucleotide sequence varying from the sequence of the nucleotide sequence specified in (a) as a result of degeneracy of the genetic code; and (c) an allelic variant of the nucleotide sequence specified in (a). 2. The polynucleotide of claim 1 wherein said nucleotide sequence encodes for a protein having a biological activity of the human IL- 1 1 receptor. 3. The polynucleotide of claim 1 wherein said nucleotide sequence is operably linked to an expression control sequence. 4 The polynucleotide of claim 2 comprising the nucleotide sequence of SEQ ID NO 1 from nucleotide 803 to nucleotide 1999 5. The polynucleotide of claim 2 comprising the nucleotide sequence of SEQ ID NO 1 from nucleotide 803 to nucleotide 1828 oi a fragment thereof 6. The polynucleotide of claim 2 comprising the nucleotide sequence of SEQ ID NO: l from nucleotide 1907 to nucleotide 1999 or a fragment thereof. 7. The polynucleotide of claim 1 comprising the nucleotide sequence of SEQ ID NO: l from nucleotide 734 to nucleotide 1999. 8. The polynucleotide of claim 1 comprising the nucleotide sequence of SEQ ID NO: l from nucleotide 1067 to nucleotide 1828. 9. The polynucleotide of claim 1 comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 1067 to nucleotide 1999. 10. A host cell transformed with the polynucleotide of claim 3. 1 1. The host cell of claim 8, wherein said cell is a mammalian cell. 12. A process for producing a human IL-l lR protein, said process comprising: (a) growing a culture of the host cell of claim 10 in a suitable culture medium; and (b) purifying the human IL-l lR protein from the culture 13. An isolated human IL-l lR 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 acids 24 to 422; (c) the amino acid sequence of SEQ ID NO:2 from amino acids 24 to 365; (d) the amino acid sequence of SEQ ID NO:2 from amino acids 391 to 422; (e) the amino acid sequence of SEQ ID NO: 2 from amino acids 102 to 422; (f) the amino acid sequence of SEQ ID NO: 2 from amino acids 102 to 365; and (g) fragments of (a)-(f) having a biological activity of the human IL-11 receptor. 14. The protein of claim 13 comprising the amino acid sequence of SEQ ID NO:2. 15. The protein of claim 13 comprising the sequence from amino acid 24 to 365 of SEQ ID NO:2. 16. A pharmaceutical composition comprising a protein of claim 13 and a pharmaceutically acceptable carrier. 17. A protein produced according to the process of claim 12. 18. A composition comprising an antibody which specifically reacts with a protein of claim 13. 19. A method of identifying an inhibitor of IL-11 binding to the human IL-11 receptor which comprises: (a) combining a protein of claim 13 with IL-11 or a fragment thereof, said combination forming a first binding mixture; (b) measuring the amount of binding between the protein and the IL-11 or fragment in the first binding mixture; (c) combining a compound with the protein and the IL-11 or fragment to form a second binding mixture; (d) measuring the amount of binding in the second binding mixture; and (e) comparing the amount of binding in the first binding mixture with the amount of binding in the second binding mixture; wherein the compound is capable of inhibiting IL-11 binding to the human IL-11 receptor when a decrease in the amount of binding of the second binding mixture occurs. 20. The method of claim 19 wherein the first and second binding mixture comprise gpl30 or a fragment thereof capable of binding to the protein of claim 13 or the IL-11 or fragment used therein. 21. An inhibitor identified by the method of claim 19. 22. A pharmaceutical composition comprising the inhibitor of claim 21 and a pharmaceutically acceptable carrier. 23. A method of inhibiting binding of IL-11 to the human IL-11 receptor in a mammalian subject, said method comprising administering a therapeutically effective amount of a composition of claim 22. 24. A method of inhibiting binding of IL-11 to the human IL-11 receptor in a mammalian subject, said method comprising administering a therapeutically effective amount of a composition of claim 16. 25. A method of inhibiting binding of IL-11 to the human IL-11 receptor in a mammalian subject, said method comprising administering a therapeutically effective amount of a composition of claim 18. 26. A method of treating or preventing loss of bone mass in a mammalian subject, said method comprising administering a therapeutically effective amount of a composition of claim 22. 27. A method of treating or preventing loss of bone mass in a mammalian subject, said method comprising administering a therapeutically effective amount of a composition of claim 16. 28. A method of treating or preventing loss of bone mass in a mammalian subject, said method comprising administering a therapeutically effective amount of a composition of claim 18. 29. An isolated polynucleotide comprising a nucleotide sequence capable of hybridizing under stringent conditions to polynucleotide of claim 4. 30. 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 acids 24 to 422; (c) the amino acid sequence of SEQ ID NO:2 from amino acids 24 to 365; (d) the amino acid sequence of SEQ ID NO:2 from amino acids 391 to 422; (e) the amino acid sequence of SEQ ID NO:2 from amino acids 112 to 422; (f) the amino acid sequence of SEQ ID NO:2 from amino acids 112 to 365; and (g) fragments of (a)-(f) having a biological activity of the human IL-1 1 receptor.

Inventors: TOBIN JAMES F, TOBIN, JAMES, F.
Applicants: GENETICS INST, GENETICS INSTITUTE, INC.
Application year: 1995
Main classification: C12N15
IPC codes: A61K38, A61P43, C07K14, C12N5, C12N15, C12P21, C12R1, G01N33