Zcytor19 polynucleotides, polypeptides, antibodies and methods of use

Abstract

Novel polypeptides, polynucleotides encoding the polypeptides, and related compositions and methods are disclosed for zcytor19, a novel class II cytokine receptor. The polypeptides may be used within methods for detecting ligands that stimulate the proliferation and/or development of hematopoietic, lymphoid and myeloid cells in vitro and in vivo. Ligand-binding receptor polypeptides can also be used to block ligand activity in vitro and in vivo. The polynucleotides encoding zcytor19, are located on chromosome 1p36.11, and can be used to identify a region of the genome associated with human disease states. The present invention also includes methods for producing the protein, uses therefor and antibodies thereto.

1 . An isolated polynucleotide that encodes a polypeptide comprising an amino acid sequence selected from the group consisting of:
 (a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 223 (Pro);   (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 226 (Asn);   (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 249 (Trp);   (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 250 (Lys) to amino acid number 491 (Arg);   (e) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 250 (Lys) to 520 (Arg);   (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 491 (Arg);   (g) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 21 (Arg) to amino acid number 520 (Arg);   (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 491 (Arg); and   (i) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 1 (Met) to amino acid number 520 (Arg);   (j) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 163 (Trp);   (k) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 211 (Ser); and   (l) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 1 (Met) to amino acid number 211 (Ser).   
     
     
         2 . An isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of:
 (a) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 61 to nucleotide 669;   (b) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 61 to 678;   (c) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 61 to nucleotide 747;   (d) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 748 to nucleotide 1473;   (e) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:18 from nucleotide 748 to nucleotide 1560;   (f) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 61 to nucleotide 1473;   (g) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:18 from nucleotide 61 to nucleotide 1560;   (h) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 1 to nucleotide 1473;   (i) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:18 from nucleotide 1 to nucleotide 1560;   (j) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:20 from nucleotide 61 to nucleotide 489;   (k) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:20 from nucleotide 61 to nucleotide 633; and   (l) a polynucleotide comprising a nucleotide sequence as shown in SEQ ID NO:20 from nucleotide 1 to nucleotide 633.   
     
     
         3 . An expression vector comprising the following operably linked elements:
 a transcription promoter;   a DNA comprising the polynucleotide of  claim 1 ; and   a transcription terminator,   wherein the promoter is operably linked to the DNA segment, and the DNA segment is operably linked to the transcription terminator.   
     
     
         4 . An expression vector according to  claim 3 , comprising a secretory signal sequence operably linked to the DNA segment. 
     
     
         5 . A cultured cell comprising an expression vector according to  claim 4 , wherein the cell expresses a polypeptide encoded by the DNA segment. 
     
     
         6 . A method of producing a polypeptide comprising:
 culturing a cell according to  claim 5 ; and   isolating the polypeptide produced by the cell.   
     
     
         7 . A cultured cell into which has been introduced an expression comprising the following operably linked elements:
 a transcription promoter;   a DNA segment encoding a polypeptide from the group consisting of:   (a) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 21 (Arg) to amino acid number 223 (Pro);   (b) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 21 (Arg) to amino acid number 226 (Asn); and   (c) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 21 (Arg) to amino acid number 249 (Trp); and   a transcription terminator,   wherein the promoter is operably linked to the DNA segment, and the DNA segment is operably linked to the transcription terminator, and wherein the cell expresses a soluble receptor polypeptide encoded by the DNA segment.   
     
     
         8 . A DNA construct encoding a fusion protein, the DNA construct comprising:
 a first DNA segment encoding a polypeptide comprising a sequence of amino acid residues selected from the group consisting of:   (a) the amino acid sequence of SEQ ID NO:2 from amino acid number 1 (Met), to amino acid number 20 (Gly);   (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 223 (Pro);   (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 226 (Asn);   (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 249 (Trp);   (e) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 250 (Lys) to amino acid number 491 (Arg);   (f) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 250 (Lys) to amino acid number 520 (Arg);   (g) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 227 (Trp) to amino acid number 249 (Trp);   (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 227 (Trp) to amino acid number 491 (Arg);   (i) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 227 (Trp) to amino acid number 520 (Arg);   (j) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 491 (Arg);   (k) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 21 (Arg) to amino acid number 520 (Arg);   (l) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 491 (Arg); and   (m) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 1 (Met) to amino acid number 520 (Arg);   (n) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 163 (Trp);   (o) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 211 (Ser); and   (p) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 1 (Met) to amino acid number 211 (Ser); and   at least one other DNA segment encoding an additional polypeptide,   wherein the first and other DNA segments are connected in-frame; and   wherein the first and other DNA segments encode the fusion protein.   
     
     
         9 . An expression vector comprising the following operably linked elements:
 a transcription promoter;   a DNA construct encoding a fusion protein according to  claim 8 ; and   a transcription terminator,   wherein the promoter is operably linked to the DNA construct, and the DNA construct is operably linked to the transcription terminator.   
     
     
         10 . A cultured cell comprising an expression vector according to  claim 9 , wherein the cell expresses a polypeptide encoded by the DNA construct. 
     
     
         11 . A method of producing a fusion protein comprising:
 culturing a cell according to  claim 10 ; and   isolating the polypeptide produced by the cell.   
     
     
         12 . An isolated polypeptide comprising a sequence of amino acid residues selected from the group consisting of:
 (a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 223 (Pro);   (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 226 (Asn);   (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 249 (Trp);   (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 250 (Lys) to amino acid number 491 (Arg);   (e) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 250 (Lys) to 520 (Arg);   (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 491 (Arg);   (g) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 21 (Arg) to amino acid number 520 (Arg);   (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 491 (Arg);   (i) the amino acid sequence as shown in SEQ ID NO:19 from amino acid number 1 (Met) to amino acid number 520 (Arg);   (j) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 163 (Trp);   (k) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 211 (Ser); and   (l) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 1 (Met) to amino acid number 211 (Ser).   
     
     
         13 . An isolated polypeptide according to  claim 12 , wherein the polypeptide consists of the sequence of amino acid residues. 
     
     
         14 . An isolated polypeptide comprising an amino acid segment selected from the group consisting of:
 (a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 223 (Pro) or amino acid number 226 (Asn);   (b) the amino acid sequence as shown in SEQ ID NO:4;   and   (c) sequences that are at least 90% identical to (a), or (b),   wherein the polypeptide is substantially free of transmembrane and intracellular domains ordinarily associated with hematopoietic receptors.   
     
     
         15 . The isolated polypeptide of  claim 14 , comprising another cytokine receptor subunit. 
     
     
         16 . A method of producing an antibody to a polypeptide comprising:
 inoculating an animal with a polypeptide selected from the group consisting of:   (a) a polypeptide consisting of 50 to 471 contiguous amino acids of SEQ ID NO:2 from amino acid number 21 (Arg), to amino acid number 491 (Arg);   (b) a polypeptide consisting of 50 to 500 contiguous amino acids of SEQ ID NO:19 from amino acid number 21 (Arg), to amino acid number 520 (Arg);   (c) a polypeptide comprising amino acid number 21 (Arg) to 119 (Tyr) of SEQ ID NO:2; and   (d) a polypeptide comprising amino acid number 125 (Pro) to 223 (Pro) of SEQ ID NO:2;   wherein the polypeptide elicits an immune response in the animal to produce the antibody; and   isolating the antibody from the animal   
     
     
         17 . The method of  claim 16 , wherein the antibody is a monoclonal antibody. 
     
     
         18 . An antibody that specifically binds to the polypeptide of  claim 12 . 
     
     
         19 . The antibody of  claim 18 , wherein the antibody is a monoclonal antibody or antibody fragment. 
     
     
         20 . The antibody of  claim 18 , wherein the antibody is a chimeric antibody. 
     
     
         21 . The antibody of  claim 18 , wherein the antibody is humanized. 
     
     
         22 . The antibody of  claim 18 , wherein the antibody is human. 
     
     
         23 . An antibody that specifically binds to a polypeptide of  claim 14 . 
     
     
         24 . An antibody that specifically binds to amino acids 65 to 70 of SEQ ID NO:2. 
     
     
         25 . A method of detecting, in a test sample, the presence of a modulator of the activity of a cytokine receptor protein comprising:
 culturing a cell into which has been introduced an expression vector according to  claim 3 , wherein the cell expresses the protein encoded by the DNA segment in the presence and absence of a test sample; and   comparing levels of activity of the protein in the presence and absence of a test sample, by a biological or biochemical assay; and   determining from the comparison, the presence of modulator the cytokine receptor protein activity in the test sample.   
     
     
         26 . A method for detecting a cytokine receptor ligand within a test sample, comprising:
 contacting a test sample with a cytokine receptor polypeptide comprising an amino acid sequence from the group consisting of:   (a) the amino acid sequence as shown in SEQ ID NO:4;   (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 21 (Arg) to amino acid number 226 (Asn); and   (c) the amino acid sequence as shown in SEQ ID NO:21 from amino acid number 21 (Arg) to amino acid number 211 (Ser); and   detecting the binding of the cytokine receptor polypeptide to a ligand in the sample.   
     
     
         27 . A method for detecting a cytokine receptor ligand according to  claim 26 , wherein the cytokine receptor polypeptide is membrane bound within a cultured cell, and the detecting step comprises measuring a biological response in the cultured cell. 
     
     
         28 . A method for detecting a genetic abnormality in a patient, comprising:
 obtaining a genetic sample from a patient;   producing a first reaction product by incubating the genetic sample with a polynucleotide comprising at least 14 contiguous nucleotides of SEQ ID NO:1 or the complement of SEQ ID NO:1, under conditions wherein said polynucleotide will hybridize to complementary polynucleotide sequence;   visualizing the first reaction product; and   comparing said first reaction product to a control reaction product from a wild type patient, wherein a difference between said first reaction product and said control reaction product is indicative of a genetic abnormality in the patient.   
     
     
         29 . A method for detecting a cancer in a patient, comprising:
 obtaining a tissue or biological sample from a patient;   incubating the tissue or biological sample with an antibody of  claim 18  under conditions wherein the antibody binds to its complementary polypeptide in the tissue or biological sample;   visualizing the antibody bound in the tissue or biological sample; and   comparing levels of antibody bound in the tissue or biological sample from the patient to a normal control tissue or biological sample,   wherein an increase in the level of antibody bound to the patient tissue or biological sample relative to the normal control tissue or biological sample is indicative of a cancer in the patient.   
     
     
         30 . A method for detecting a cancer in a patient, comprising:
 obtaining a tissue or biological sample from a patient;   labeling a polynucleotide comprising at least 14 contiguous nucleotides of SEQ ID NO:1, SEQ ID NO:18 or SEQ ID NO:20 or the complement of SEQ ID NO:1, SEQ ID NO:18 or SEQ ID NO:20;   incubating the tissue or biological sample with under conditions wherein the polynucleotide will hybridize to complementary polynucleotide sequence;   visualizing the labeled polynucleotide in the tissue or biological sample; and   comparing the level of labeled polynucleotide hybridization in the tissue or biological sample from the patient to a normal control tissue or biological sample,   wherein an increase in the labeled polynucleotide hybridization to the patient tissue or biological sample relative to the normal control tissue or biological sample is indicative of a cancer in the patient.