Human receptor proteins; related reagents and methods

Abstract

Nucleic acids encoding mammalian, e.g., human receptors, purified receptor proteins and fragments thereof. Antibodies, both polyclonal and monoclonal, are also provided. Methods of using the compositions for both diagnostic and therapeutic utilities are provided.

What is claimed is:  
     
         1 . An isolated or recombinant IL-1RD9 polypeptide: 
 a) consisting of SEQ ID NO: 6, 8, 10, 12, 14, or 16;    b) encoded by a polynucleotide comprising the open reading frame of SEQ ID NO: 5, 7, 9, 11, 13, or 15; or    c) encoded by a naturally occurring allelic variant of a polynucleotide comprising the open reading frame of SEQ ID NO: 5, 7, 9, 11, 13, or 15.    
     
     
         2 . The polypeptide of  claim 1 , encoded by a naturally occurring allelic variant of a polynucleotide comprising the open reading frame of SEQ ID NO: 5, 7, 9, 11, 13, or 15.  
     
     
         3 . An isolated or recombinant IL-1RD9 polypeptide which: 
 a) has an apparent molecular weight of approximately 68.3 kD as determined by calculation from sequence and an estimated pI of approximately 9.04; and    b) is found on T cells;    wherein said polypeptide has at least one of the following properties: 
 i) is a heterodimer;  
 iii) is an IL-1 receptor α subunit type, or  
 iii) when brought into contact with IL-1RD5 and IL-1α, for a sufficient time, forms a functional high affinity receptor complex that activates an NFκB transcription factor reporter construct.  
   
     
     
         4 . An isolated or recombinant polypeptide comprising a segment of contiguous amino acid residues selected from the following group: 
 a) 15 contiguous amino acid residues of said polypeptide of  claim 2;     b) 20 contiguous amino acid residues of said polypeptide of  claim 2;     c) 25 contiguous amino acid residues of said polypeptide of  claim 2;     d) 30 contiguous amino acid residues of said polypeptide of  claim 2;     e) 35 contiguous amino acid residues of said polypeptide of  claim 2;  or    f) 40 contiguous amino acid residues of said polypeptide of  claim 2 .    
     
     
         5 . The polypeptide of  claim 1  which is immunogenic.  
     
     
         6 . An isolated or recombinant polypeptide comprising an immunogenic peptide of said polypeptide of  claim 3 .  
     
     
         7 . An isolated or recombinant polypeptide comprising an immunogenic polypeptide of  claim 4 .  
     
     
         8 . A fusion protein comprising said polypeptide of  claim 4  and: 
 a) a detection or purification tag selected from the group consisting of a FLAG, His6, and immunoglobulin peptide;  
 b) a carrier protein selected from the group consisting of keyhole limpet hemocyanin, bovine serum albumin, and tetanus toxoid; or  
 c) another peptide selected from the group consisting of luciferase, bacterial β-galactosidase, trpE, protein A, β-lactamase, alpha amylase, alcohol dehydrogenase, and yeast alpha mating factor.  
 
     
     
         9 . A fusion protein comprising said polypeptide of  claim 5  and: 
 a) a detection or purification tag selected from the group consisting of a FLAG, His6, and immunoglobulin peptide;  
 b) a carrier protein selected from the group consisting of keyhole limpet hemocyanin, bovine serum albumin, the tetanus toxoid; or  
 c) another peptide selected from the group consisting of luciferase, bacterial β-galactosidase, trpE, protein A, β-lactamase, alpha amylase, alcohol dehydrogenase, and yeast alpha mating factor.  
 
     
     
         10 . A composition comprising said polypeptide of  claim 1 , that is: 
 a) in a pharmaceutically acceptable carrier;    b) in a sterile composition;    c) in a buffered solution;    d) in an aqueous suspension; or    e) in combination with IL-1RD5 and/or IL-1α.    
     
     
         11 . A composition comprising said polypeptide of  claim 4 , that is: 
 a) in a pharmaceutically acceptable carrier;    b) in a sterile composition;    c) in a buffered solution; or    d) in an aqueous suspension.    
     
     
         12 . A polypeptide of  claim 4 , that is: 
 a) denatured;    b) immunopurified;    c) attached to a solid substrate;    d) detectably labeled; or    e) chemically synthesized.    
     
     
         13 . A polypeptide of  claim 5 , that is: 
 a) denatured;    b) immunopurified;    c) attached to a solid substrate;    d) detectably labeled; or    e) chemically synthesized.    
     
     
         14 . A kit comprising said polypeptide of  claim 1 , and: 
 a) a compartment comprising said protein; or    b) instructions for use or disposal of reagents in said kit.    
     
     
         15 . A kit comprising said polypeptide of  claim 4 , and: 
 a) a compartment comprising said protein; or    b) instructions for use or disposal of reagents in said kit.    
     
     
         16 . A method of raising an antibody, comprising immunizing an animal with a polypeptide of  claim 5 .  
     
     
         17 . A method of producing an antibody:antigen complex, comprising contacting a polypeptide of  claim 5  with an antibody which specifically binds said polypeptide, thereby forming said complex.  
     
     
         18 . A composition of matter selected from the group consisting of: 
 a) a substantially pure or recombinant IL-1RD8 polypeptide exhibiting identity over a length of at least about 12 amino acids to SEQ ID NO: 4;    b) a natural sequence IL-1RD8 comprising SEQ ID NO: 4;    c) a fusion polypeptide comprising IL-1RD8 sequence;    d) a substantially pure or recombinant IL-1RD10 polypeptide exhibiting identity over a length of at least about 12 amino acids to SEQ ID NO: 35;    e) a natural sequence IL-1RD10 comprising SEQ ID NO: 35; and    f) a fusion protein comprising IL-1RD10 sequence.    
     
     
         19 . A substantially pure or isolated polypeptide comprising a segment exhibiting sequence identity to a corresponding portion of an: 
 a) IL-1RD8 of  claim 18 , wherein: 
 i) said polypeptide further exhibits identity to a distinct segment of 9 amino acids;  
 ii) said length of identity is at least 17 amino acids;  
 iii) said length of identity is at least about 25 amino acids; or  
   b) IL-1RD10 of  claim 18 , wherein: 
 i) said polypeptide further exhibits identity to a distinct segment of 9 amino acids;  
 ii) said length of identity is at least 17 amino acids;  
 iii) said length of identity is at least about 25 amino acids.  
   
     
     
         20 . The composition of matter of  claim 18 , wherein said: 
 a) IL-1RD8 comprises a mature sequence of Table 1;    b) IL-1RD10 comprises a mature sequence of Table 3; or    c) polypeptide: 
 i) is from a warm blooded animal selected from a primate, such as a human;  
 ii) comprises at least one polypeptide segment of SEQ ID NO: 4 or 35;  
 iii) exhibits a plurality of portions exhibiting said identity;  
 iv) is a natural allelic variant of a primate or rodent IL-1RD8 or primate IL-1RD10;  
 v) has a length at least about 30 amino acids;  
 vi) exhibits at least two non-overlapping epitopes which are specific for a primate or rodent IL-1RD8 or primate IL-1RD10;  
 vii) exhibits sequence identity over a length of at least about 20 amino acids to a primate IL-1RD8 or IL-1RD10;  
 viii) has a molecular weight of at least 100 kD with natural glycosylation;  
 ix) is a synthetic polypeptide;  
 x) is attached to a solid substrate;  
 xi) is conjugated to another chemical moiety;  
 xii) is a 5-fold or less substitution from natural sequence; or  
 xiii) is a deletion or insertion variant from a natural sequence.  
   
     
     
         21 . A composition comprising: 
 a) a sterile IL-1RD8 polypeptide of  claim 18;     b) said IL-1RD8 polypeptide of  claim 18  and a carrier, wherein said carrier is: 
 i) an aqueous compound, including water, saline, and/or buffer; and/or  
 ii) formulated for oral, rectal, nasal, topical, or parenteral administration;  
   c) a sterile IL-1RD10 polypeptide of  claim 18;  or    d) said IL-1RD10 polypeptide of  claim 18  and a carrier, wherein said carrier is: 
 i) an aqueous compound, including water, saline, and/or buffer; and/or  
 ii) formulated for oral, rectal, nasal, topical, or parenteral administration.  
   
     
     
         22 . A fusion protein of  claim 18 , comprising: 
 a) mature protein sequence of Table 1 or 3;    b) a detection or purification tag, including a FLAG, His6, or Ig sequence; or    c) sequence of another receptor protein.    
     
     
         23 . A kit comprising a polypeptide of  claim 18 , and: 
 a) a compartment comprising said polypeptide; and/or    b) instructions for use or disposal of reagents in said kit.    
     
     
         24 . A binding compound comprising an antigen binding site from an antibody, which specifically binds to a natural: 
 A) IL-1RD8 polypeptide of  claim 18 , wherein: 
 a) said polypeptide is a primate or rodent protein;  
 b) said binding compound is an Fv, Fab, or Fab2 fragment;  
 c) said binding compound is conjugated to another chemical moiety; or  
 d) said antibody: 
 i) is raised against a polypeptide sequence of a mature polypeptide of Table 1;  
 ii) is raised against a mature primate or rodent IL-1RD8;  
 iii) is raised to a purified human IL-1RD8;  
 iv) is raised to a purified mouse IL-1RD8;  
 v) is immunoselected;  
 vi) is a polyclonal antibody;  
 vii) binds to a denatured IL-1RD8;  
 viii) exhibits a Kd to antigen of at least 30 μM;  
 ix) is attached to a solid substrate, including a bead or plastic membrane;  
 x) is in a sterile composition; or  
 xi) is detectably labeled, including a radioactive or fluorescent label; or  
 
   B) IL-1RD10 polypeptide of  claim 18 , wherein: 
 a) said polypeptide is a primate polypeptide;  
 b) said binding compound is an Fv, Fab, or Fab2 fragment;  
 c) said binding compound is conjugated to another chemical moiety;  
 d) said polypeptide is associated with an IL-1 receptor alpha type subunit; or  
 e) said antibody: 
 i) is raised against a peptide sequence of a mature polypeptide of Table 3;  
 ii) is raised against a mature primate IL-1RD10;  
 iii) is raised to a purified human IL-1RD10;  
 iv) is immunoselected;  
 v) is a polyclonal antibody;  
 vi) binds to a denatured IL-1RD10;  
 vii) exhibits a Kd to antigen of at least 30 μM;  
 viii) is attached to a solid substrate, including a bead or plastic membrane;  
 ix) is in a sterile composition; or  
 x) is detectably labeled, including a radioactive or fluorescent label  
 
   
     
     
         25 . A kit comprising said binding compound of  claim 24 , and: 
 a) a compartment comprising said binding compound; and/or    b) instructions for use or disposal of reagents in said kit.    
     
     
         26 . A method of: 
 A) making an antibody of  claim 23 , comprising immunizing an immune system with an immunogenic amount of: 
 a) a primate IL-1RD8 polypeptide; or  
 b) a primate IL-1RD10 polypeptide;  
   thereby causing said antibody to be produced; or    B) producing an antigen:antibody complex, comprising contacting: 
 a) a primate IL-1RD8 polypeptide with an antibody of claim  23 A; or  
 b) a primate IL-1RD10 polypeptide with an antibody of claim  23 B;  
   thereby allowing said complex to form.    
     
     
         27 . A composition comprising: 
 a) a sterile binding compound of  claim 23 , or    b) said binding compound of  claim 23  and a carrier, wherein said carrier is: 
 i) an aqueous compound, including water, saline, and/or buffer; and/or  
 ii) formulated for oral, rectal, nasal, topical, or parenteral administration.  
   
     
     
         28 . An isolated or recombinant nucleic acid encoding a protein or peptide or fusion protein of  claim 18 , wherein: 
 a) said IL-1RD8 or IL-1RD10 is from a mammal; or    b) said nucleic acid: 
 i) encodes an antigenic polypeptide sequence of Table 1 or 3;  
 ii) encodes a plurality of antigenic polypeptide sequences of Table 1 or 3;  
 iii) exhibits identity to a natural cDNA encoding said segment;  
 iv) is an expression vector;  
 v) further comprises an origin of replication;  
 vi) is from a natural source;  
 vii) comprises a detectable label;  
 viii) comprises synthetic nucleotide sequence;  
 ix) is less than 6 kb, preferably less than 3 kb;  
 x) is from a mammal, including a primate, such as a human;  
 xi) comprises a natural full length coding sequence;  
 xii) is a hybridization probe for a gene encoding said IL-1RD8 or IL-1RD10;  
 xiii) comprises a plurality of nonoverlapping segments of at least 15 nucleotides from Table 1 or 3; or  
 xiv) is a PCR primer, PCR product, or mutagenesis primer.  
   
     
     
         29 . A cell transfected or transformed with a recombinant nucleic acid of  claim 28 .  
     
     
         30 . The cell of  claim 29 , wherein said cell is: 
 a) a prokaryotic cell;    b) a eukaryotic cell;    c) a bacterial cell;    d) a yeast cell;    e) an insect cell;    f) a mammalian cell;    g) a mouse cell;    h) a primate cell; or    i) a human cell.    
     
     
         31 . A kit comprising said nucleic acid of  claim 28 , and: 
 a) a compartment comprising said nucleic acid;    b) a compartment further comprising a primate or rodent IL-1RD8 or primate IL-1RD10 polypeptide; and/or    c) instructions for use or disposal of reagents in said kit.    
     
     
         32 . A method of: 
 A) making a polypeptide, comprising expressing said nucleic acid of  claim 28 , thereby producing said polypeptide; or    B) making a duplex nucleic acid, comprising contacting said nucleic acid of  claim 28  with a hybridizing nucleic acid, thereby allowing said duplex to form.    
     
     
         33 . A nucleic acid which: 
 a) hybridizes under wash conditions of 40° C. and less than 2M salt to SEQ ID NO: 3, 19, or 34; or    b) exhibits identity over a stretch of at least about 30 nucleotides to a primate IL-1RD8 or IL-1RD10.    
     
     
         34 . The nucleic acid of  claim 33 , wherein: 
 a) said wash conditions are at 55° C. and/or 500 mM salt; or    b) said stretch is at least 55 nucleotides.    
     
     
         35 . The nucleic acid of  claim 34 , wherein: 
 a) said wash conditions are at 65° C. and/or 150 mM salt; or    b) said stretch is at least 75 nucleotides.    
     
     
         36 . A method of modulating physiology or development of a cell or tissue culture cells comprising contacting said cell with an agonist or antagonist of a primate IL-1RD8 or IL-1RD10.  
     
     
         37 . The method of  claim 36 , wherein said cell is transformed with a nucleic acid encoding either an IL-1RD8 or IL-1RD10, and another IL-1R.