Zcytor19 heterodimeric cytokine receptor polynucleotides, polypeptides, antibodies and methods

Abstract

Novel methods are disclosed for forming a heterodimeric receptor complex with IL-28R and CRF2-4. The methods may be used for detecting and treating viral infections in in vitro and in vivo. Ligand-binding receptor polypeptides can also be used to block ligand activity in vitro and in vivo. The present invention also includes methods for producing the protein, uses therefor and antibodies thereto.

1 . An isolated heterodimeric receptor comprising a first subunit and a second subunit wherein the first subunit comprises a polypeptide having 90% sequence identity to amino acid residues 21 to 226 as shown in SEQ ID NO:2 and the second subunit comprises the extracellular domain of the amino acid sequence as shown in SEQ ID NO:64. 
     
     
         2 . The heterodimeric receptor of  claim 1 , wherein the heterodimeric receptor specifically binds a polypeptide selected from the group consisting of:
 a) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO:52;   b) a polypeptide comprising amino acid residues 20-200 of SEQ ID NO:55;   c) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO: 57   d) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:60; and   e) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:62.   
     
     
         3 . The heterodimeric receptor of  claim 1 , wherein the first subunit comprises the amino acid sequence as shown in SEQ ID NO: 2 from amino acid residue 21 to amino acid residue 491. 
     
     
         4 . An expression vector encoding the heterodimeric receptor of  claim 1 , wherein the expression vector comprises a transcription promoter, a first DNA segment comprising a polynucleotide sequence encoding the first subunit, a second DNA segment comprising a polynucleotide sequence encoding the second subunit, and a transcription terminator. 
     
     
         5 . An isolated cell comprising the expression vector of  claim 4 . 
     
     
         6 . An isolated heterodimeric receptor comprising a first subunit and a second subunit wherein the first subunit comprises a polypeptide having 90% sequence identity to amino acid residues 21 to 520 as shown in SEQ ID NO:19 and the second subunit comprises the extracellular domain of the amino acid sequence as shown in SEQ ID NO:64. 
     
     
         7 . The heterodimeric receptor of  claim 6 , wherein the heterodimeric receptor specifically binds a polypeptide selected from the group consisting of:
 a) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO:52;   b) a polypeptide comprising amino acid residues 20-200 of SEQ ID NO:55;   c) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO: 57   d) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:60; and   e) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:62.   
     
     
         8 . The heterodimeric receptor of  claim 6 , wherein binding of the ligand to the heterodimeric receptor induces signal transduction in a cell expressing the heterodimeric receptor. 
     
     
         9 . An expression vector encoding the heterodimeric receptor of  claim 6 , wherein the expression vector comprises a transcription promoter, a first DNA segment comprising a polynucleotide sequence encoding the first subunit, a second DNA segment comprising a polynucleotide sequence encoding the second subunit, and a transcription terminator. 
     
     
         10 . An isolated cell comprising the expression vector of  claim 9 . 
     
     
         11 . An isolated heterodimeric receptor comprising a first subunit and a second subunit wherein the first subunit comprises a polypeptide having 90% sequence identity to amino acid residues 21 to 211 as shown in SEQ ID NO:21 and the second subunit comprises the extracellular domain of the amino acid sequence as shown in SEQ ID NO:64. 
     
     
         12 . The heterodimeric receptor of  claim 11 , wherein the heterodimeric receptor specifically binds a polypeptide selected from the group consisting of:
 a) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO:52;   b) a polypeptide comprising amino acid residues 20-200 of SEQ ID NO:55;   c) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO: 57   d) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:60; and   e) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:62.   
     
     
         13 . An expression vector encoding the heterodimeric receptor of  claim 11 , wherein the expression vector comprises a transcription promoter, a first DNA segment comprising a polynucleotide sequence encoding the first subunit, a second DNA segment comprising a polynucleotide sequence encoding the second subunit, and a transcription terminator. 
     
     
         14 . An isolated cell comprising the expression vector of  claim 13 . 
     
     
         15 . An isolated heterodimeric receptor comprising a first subunit and a second subunit wherein the first subunit comprises a polypeptide selected from:
 a) a polypeptide comprising the amino acid sequence as shown in SEQ ID NO:21 from amino acid residue 21 to amino acid residue 211;   b) a polypeptide comprising the amino acid sequence as shown in SEQ ID NO:2 from amino acid residue 21 to amino acid residue 226;   and wherein the second subunit comprises the extracellular domain of the polypeptide as shown in SEQ ID NO: 64.   
     
     
         16 . An isolated heterodimeric receptor comprising a first subunit and a second subunit wherein the first subunit comprises a polypeptide selected from:
 a) a polypeptide comprising the amino acid sequence as shown in SEQ ID NO:2 from amino acid residue 21 to amino acid residue 491;   b) a polypeptide comprising the amino acid sequence as shown in SEQ ID NO:19 from amino acid residue 21 to amino acid residue 520;   and wherein the second subunit comprises the polypeptide as shown in SEQ ID NO: 64.   
     
     
         17 . An isolated polypeptide comprising a sequence of amino acid residues that is at least 90% identical to the amino acid sequence as shown in SEQ ID NO:21 from amino acid residue 21 to amino acid residue 163, as shown in SEQ ID NO:21 from amino acid residue 1 to amino acid residue 163, as shown in SEQ ID NO:21 from amino acid residue 21 to amino acid residue 211, or as shown in SEQ ID NO:21 from amino acid residue 1 to amino acid residue 211. 
     
     
         18 . The polypeptide of  claim 16 , wherein the polypeptide specifically binds a ligand comprising a polypeptide selected from the group consisting of:
 a) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO:52;   b) a polypeptide comprising amino acid residues 20-200 of SEQ ID NO:55;   c) a polypeptide comprising amino acid residues 22-205 of SEQ ID NO: 57   d) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:60; and   e) a polypeptide comprising amino acid residues 29-202 of SEQ ID NO:62.   
     
     
         19 . An in vitro method for detecting a ligand in a sample, wherein the ligand comprises a polypeptide selected from the group consisting of:
 (a) amino acid residues 22-205 of SEQ ID NO: 52,   (b) amino acid residues 20-200 of SEQ ID NO: 55,   (c) amino acid residues 22-205 of SEQ ID NO: 57,   (d) amino acid residues 29-202 of SEQ ID NO: 60, and   (e) amino acid residues 29-202 of SEQ ID NO: 62;   wherein said method comprises contacting the sample with a heterodimeric receptor comprising a first receptor polypeptide and a second receptor polypeptide;   wherein said first receptor polypeptide comprises a sequence of amino acid residues that is at least 90% identical to:   (i) amino acids 21-223 of SEQ ID NO: 2;   (ii) amino acids 21-226 of SEQ ID NO: 2;   (iii) amino acids 21-223 of SEQ ID NO: 19; or   (iv) amino acids 21-226 of SEQ ID NO: 19;   wherein said second receptor polypeptide comprises the extracellular domain of CRF2-4;   and wherein said first heterodimeric receptor polypeptide binds said ligand;   and detecting binding of said ligand to said heterodimeric receptor.   
     
     
         20 . The method according to  claim 19 , wherein said first receptor polypeptide comprises a sequence of amino acid residues that is at least 90% identical to:
 (i) amino acids 250-491 of SEQ ID NO: 2; or   (ii) amino acids 250-520 of SEQ ID NO: 19.   
     
     
         21 . The method according to  claim 19 , wherein said first receptor polypeptide is fused to a heavy chain of IgGγ1. 
     
     
         22 . The method according to  claim 19 , wherein said second receptor polypeptide is fused to a human kappa light chain. 
     
     
         23 . The method according to  claim 19 , wherein said ligand is labeled, and binding of said ligand is detected by detecting said label. 
     
     
         24 . The method according to  claim 19 , wherein said receptor is labeled, and binding of said ligand is detected by detecting said label. 
     
     
         25 . The method according to  claim 19 , wherein binding of said ligand is detected by detecting an activity of said ligand, wherein said activity is selected from the group consisting of proliferation activity and anti-viral activity. 
     
     
         26 . The method according to  claim 19 , comprising isolating the ligand from the sample. 
     
     
         27 . A method according to  claim 26 , wherein said receptor is immobilised on a solid support. 
     
     
         28 . An in vitro method for antagonising an activity of a ligand, wherein the ligand comprises a polypeptide selected from the group consisting of:
 (a) amino acid residues 22-205 of SEQ ID NO: 52,   (b) amino acid residues 20-200 of SEQ ID NO: 55,   (c) amino acid residues 22-205 of SEQ ID NO: 57,   (d) amino acid residues 29-202 of SEQ ID NO: 60, and   (e) amino acid residues 29-202 of SEQ ID NO: 62;   wherein said method comprises contacting said ligand with a soluble heterodimeric receptor comprising a first receptor subunit and a second receptor subunit; and   wherein said first receptor subunit comprises a sequence of amino acid residues that is at least 90% identical to:   (i) amino acids 21-223 of SEQ ID NO: 2;   (ii) amino acids 21-226 of SEQ ID NO: 2;   (iii) amino acids 21-223 of SEQ ID NO: 19; or   (iv) amino acids 21-226 of SEQ ID NO: 19;   wherein said second receptor subunit comprises the extracellular domain of CRF2-4;   and wherein said first heterodimeric receptor subunit binds said ligand;   and wherein said activity is selected from proliferation activity and anti-viral activity.