Mammalian cell surface antigens; related reagents

Abstract

Purified genes encoding a T cell surface antigen from a mammal, reagents related thereto including purified proteins, specific antibodies, and nucleic acids encoding this antigen are provided. Methods of using said reagents and diagnostic kits are also provided.

1 . A composition of matter selected from the group consisting of: 
 a) a substantially pure or recombinant 499E9 protein or peptide exhibiting at least about 85% sequence identity over a length of at least about 12 amino acids to SEQ ID NO: 2;    b) a natural sequence 499E9 of SEQ ID NO: 2; or    c) a fusion protein comprising 499E9 sequence.    
     
     
         2 . A substantially pure or isolated protein comprising a segment exhibiting sequence identity to a corresponding portion of a 499E9 of  claim 1 , wherein: 
 a) said homology is at least about 90% identity and said portion is at least about 9 amino acids;    b) said homology is at least about 80% identity and said portion is at least about 17 amino acids; or    c) said homology is at least about 70% identity and said portion is at least about 25 amino acids.    
     
     
         3 . The composition of matter of  claim 1 , wherein said: 
 a) 499E9 comprises a mature sequence of Table 1; or    b) protein or peptide: 
 i) is from a warm blooded animal selected from a mammal, including a rodent;  
 ii) comprises at least one polypeptide segment of SEQ ID NO: 2;  
 iii) exhibits a plurality of portions exhibiting said identity;  
 iv) is a natural allelic variant of 499E9;  
 v) has a length at least about 30 amino acids;  
 vi) exhibits at least two non-overlapping epitopes which are specific for a mammalian 499E9;  
 vii) exhibits a sequence identity at least about 90% over a length of at least about 20 amino acids to a rodent 499E9;  
 viii) exhibits at least two non-overlapping epitopes which are specific for a rodent 499E9;  
 ix) exhibits a sequence identity at least about 90% over a length of at least about 20 amino acids to a rodent 499E9;  
 x) is glycosylated;  
 xi) is a synthetic polypeptide;  
 xii) is attached to a solid substrate;  
 xiii) is conjugated to another chemical moiety;  
 xiv) is a 5-fold or less substitution from natural sequence; or  
 xv) is a deletion or insertion variant from a natural sequence.  
   
     
     
         4 . A composition comprising: 
 a) a sterile 499E9 protein or peptide of  claim 1;  or    b) said 499E9 protein or peptide of  claim 1  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.  
   
     
     
         5 . The fusion protein of  claim 1 , comprising: 
 a) mature protein comprising sequence of Table 1;    b) a detection or purification tag, including a FLAG, His6, or Ig sequence; or    c) sequence of another TNF ligand protein.    
     
     
         6 . A kit comprising a protein or polypeptide of  claim 1 , and: 
 a) a compartment comprising said protein or polypeptide; and/or    b) instructions for use or disposal of reagents in said kit.    
     
     
         7 . A binding compound comprising an antigen binding portion from an antibody, which specifically binds to a natural 499E9 protein of  claim 1 , wherein: 
 a) said protein is a 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 peptide sequence of a mature polypeptide comprising sequence of Table 1;  
 ii) is raised against a mature 499E9;  
 iii) is raised to a purified 499E9;  
 iv) is immunoselected;  
 v) is a polyclonal antibody;  
 vi) binds to a denatured 499E9;  
 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.  
   
     
     
         8 . A kit comprising said binding compound of  claim 7 , and: 
 a) a compartment comprising said binding compound; and/or    b) instructions for use or disposal of reagents in said kit.    
     
     
         9 . A composition comprising: 
 a) a sterile binding compound of  claim 7;  or    b) said binding compound of  claim 7  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.  
   
     
     
         10 . A method of purifying a 499E9 protein or peptide from other materials in a mixture comprising contacting said mixture to an antibody of  claim 7 , and separating bound 499E9 from other materials.  
     
     
         11 . An isolated or recombinant nucleic acid encoding a protein or peptide or fusion protein of  claim 1 , wherein: 
 a) said 499E9 protein is from a mammal, including a rodent; or    b) said nucleic acid: 
 i) encodes an antigenic peptide sequence of Table 1;  
 ii) encodes a plurality of antigenic peptide sequences of Table 1;  
 iii) exhibits at least about 80% 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 rodent;  
 xi) comprises a natural full length coding sequence;  
 xii) is a hybridization probe for a gene encoding said TNF-ligand family protein; or  
 xiii) is a PCR primer, PCR product, or mutagenesis primer.  
   
     
     
         12 . A cell or tissue comprising a recombinant nucleic acid of  claim 11 .  
     
     
         13 . The cell of  claim 12 , 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 rodent cell; or    i) a human cell.    
     
     
         14 . A kit comprising said nucleic acid of  claim 11 , and: 
 a) a compartment comprising said nucleic acid;    b) a compartment further comprising a 499E9 protein or polypeptide; and/or    c) instructions for use or disposal of reagents in said kit.    
     
     
         15 . A nucleic acid which: 
 a) hybridizes under wash conditions of 30° C. and less than 2M salt to SEQ ID NO: 1; or    b) exhibits at least about 85% identity over a stretch of at least about 30 nucleotides to a rodent 499E9.    
     
     
         16 . The nucleic acid of  claim 15 , wherein: 
 a) said wash conditions are at 45° C. and/or 500 mM salt; or    b) said identity is at least 90% and/or said stretch is at least 55 nucleotides.    
     
     
         17 . The nucleic acid of  claim 16 , wherein: 
 a) said wash conditions are at 55° C. and/or 150 mM salt; or    b) said identity is at least 95% and/or said stretch is at least 75 nucleotides.    
     
     
         18 . A method of modulating physiology or development of a cell or tissue culture cells comprising introducing into said cell an agonist or antagonist of a 499E9 of  claim 1 .  
     
     
         19 . A method of modulating the physiology of a cell comprising contacting said cell with: 
 a) a substantially pure 499E9 or fragment of  claim 1;     b) an antibody or binding partner which specifically binds a 499E9; or    c) a nucleic acid encoding a 499E9 or peptide.    
     
     
         20 . The method of  claim 19 , wherein said cell is a T cell and said modulating of physiology is: 
 a) apoptosis of said T cell; or    b) activation of said T cell