Family of immunoregulators designated leukocyte immunoglobulin-like receptors (LIR)

Abstract

A new family of immunoreceptor molecules of the immunoglobulin superfamily, (LIR) polypeptides is described. Disclosed are sequences encoding LIR family members and their deduced amino acid sequences, polypeptides encoded by DNA that hybridize to oligonucleotide probes having defined sequences, processes for producing polypeptides of the LIR family, and antagonistic antibodies to LIR family members. LIR family members can be used to treat autoimmune diseases and disease states associated with suppressed immune function.

What is claimed is:  
     
         1 . An isolated DNA encoding an LIR polypeptide, wherein said LIR polypeptide comprises amino acid sequences selected from the group consisting of: 
 a) amino acids 5 to 50 of SEQ ID NO:2; and    b) an amino acid sequence that is at least 77% identical to the sequence of a).    
     
     
         2 . DNA encoding an LIR polypeptide, wherein said LIR polypeptide comprises the amino acid sequence: 
 Leu Xaa a  Leu Ser Xaa b  Xaa c  Pro Arg Thr Xaa d  Xaa e  Gln Xaa f  Gly Xaa g  Xaa h  Pro Xaa i  Pro Thr Leu Trp Ala Glu Pro Xaa j  Ser Phe Ile Xaa j  Xaa 70  Ser Asp Pro Lys Leu Xaa k  Leu Val Xaa m  Thr Gly    where Xaa a  is Gly or Arg; Xaa b  is Leu or Val; Xaa c  is Gly or Asp; Xaa d  is His Arg or Cys; Xaa e  is Val or Met; Xaa f  is Ala or Thr; Xaa g  is His Pro or Thr, Xaa h  Leu Ile or Phe; Xaa i  is Gly Asp or Ala; Xaa j  is Thr Ile Ser or Ala; Xaa k  is Gly or Val; Xaa m  is Met or Ala; and Xaa 70  is a sequence of 70 amino acids.    
     
     
         3 . DNA encoding an LIR polypeptide, wherein said LIR polypeptide is encoded by DNA selected from the group consisting of: 
 a) DNA capable of hybridizing under highly stringent conditions to a probe consisting essentially of nucleotides 310 to 1684 of SEQ ID NO:1; and    b) DNA capable of hybridizing under highly stringent conditions to DNA complementary to the probe of a), 
 wherein the highly stringent conditions comprise a hybridizing temperature of at least 63° C.  
   
     
     
         4 . An isolated DNA encoding an LIR polypeptide, wherein said LIR polypeptide comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of: 
 a) SEQ ID NO:2;    b) SEQ ID NO:4;    c) SEQ ID NO:8;    d) SEQ ID NO:10;    e) SEQ ID NO:12;    f) SEQ ID NO:14;    g) SEQ ED NO:16;    h) SEQ ID NO:18;    i) SEQ D NO:20; and    j) SEQ ID NO:22.    
     
     
         5 . An isolated DNA of  claim 4 , wherein said LIR polypeptide comprises an amino acid sequence selected from the group consisting of: 
 a) SEQ ID NO:2;    b) SEQ ID NO:4;    c) SEQ ID NO:8;    d) SEQ ID NO:10;    e) SEQ ID NO:12;    f) SEQ ID NO:14;    g) SEQ ID NO:16;    h) SEQ ID NO:18;    i) SEQ ID NO:20; and    j) SEQ ID NO:22.    
     
     
         6 . An isolated DNA encoding a soluble LIR polypeptide, wherein said soluble LIR polypeptide comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of: 
 a) the extracellular domain of LIR family members comprising sequences selected from the group consisting of: 
 amino acids x 1  to 458 of SEQ ID NO:2, wherein x 1  is amino acid 1 or 17;  
 amino acids x 2  to 459 of SEQ ID NO:4, wherein x 2  is amino acid 1 or 17;  
 amino acids x 3  to 439 of SEQ ID NO:8, wherein x 3  is amino acids 1 or 17;  
 amino acids x 4  to 458 of SEQ ID NO:10, wherein x 4  is amino acid 1 or 17;  
 amino acids x 5  to 241 of SEQ ID NO:12, wherein x 5  is amino acid 1 or 17;  
 amino acids x 6  to 461 of SEQ ID NO:14, wherein x 6  is amino acid 1 or 17;  
 amino acids x 7  to 449 of SEQ ED NO:16, wherein x 7  is amino acid 1 or 17;  
 amino acids x 8  to 259 of SEQ ID NO:18, wherein x 8  is amino acid 1 or 17;  
 amino acids x 9  to 443 of SEQ ID NO:20, wherein x 9  is amino acid 1 or 17; and,  
 amino acids x 10  to 456 of SEQ ID NO:22, wherein x 10  is amino acid 1 or 17;  
   b) a fragment of any of the LIR extracellular domains of a), wherein said soluble LIR polypeptide binds an MHC molecule.    
     
     
         7 . A DNA of  claim 6 , wherein said soluble LIR polypeptide comprises an amino acid sequence selected from the group consisting of: 
 a) the extracellular domain of LIR family members comprising sequences selected from the group consisting of: 
 amino acids x 1  to 458 of SEQ ID NO:2, wherein x 1  is amino acid 1 or 17;  
 amino acids x 2  to 459 of SEQ ID NO:4, wherein x 2  is amino acid 1 or 17;  
 amino acids x 3  to 439 of SEQ ID NO:8, wherein x 3  is amino acids 1 or 17;  
 amino acids x 4  to 458 of SEQ ID NO:10, wherein x 4  is amino acid 1 or 17;  
 amino acids x 5  to 241 of SEQ ID NO:12, wherein x 5  is amino acid 1 or 17;  
 amino acids x 6  to 461 of SEQ ID NO:14, wherein x 6  is amino acid 1 or 17;  
 amino acids x 7  to 449 of SEQ ID NO:16, wherein x 7  is amino acid 1 or 17;.  
 amino acids x 8  to 259 of SEQ ID NO:18, wherein x 8  is amino acid 1 or 17;  
 amino acids x 9  to 443 of SEQ ID NO:20, wherein x 9  is amino acid 1 or 17; and,  
 amino acids x 10  to 456 of SEQ ID NO:22, wherein x 10  is amino acid 1 or 17;  
   b) a fragment of any of the LIR extracellular domains of a).    
     
     
         8 . An LIR polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) amino acids 5 to 50 of SEQ ID NO:2; and    b) an amino acid sequence that is at least 77% identical to the sequence of a).    
     
     
         9 . An LIR polypeptide comprising the amino acid sequence: 
 Leu Xaa a  Leu Ser Xaa b  Xaa c  Pro Arg Thr Xaa d  Xaa e  Gln Xaa f  Gly Xaa g  Xaa h  Pro Xaa i  Pro Thr Leu Trp Ala Glu Pro Xaa j  Ser Phe Ile Xaa j  Xaa 70  Ser Asp Pro Lys Leu Xaa k  Leu Val Xaa m  Thr Gly    where Xaa a  is Gly or Arg; Xaa b  is Leu or Val; Xaa c  is Gly or Asp; Xaa d  is His Arg or Cys; Xaa e  is Val or Met; Xaa f  is Ala or Thr, Xaa g  is His Pro or Thr, Xaa h  Leu Ile or Phe; Xaa i  is Gly Asp or Ala; Xaa j  is Thr Ile Ser or Ala; Xaa k  is Gly or Val; Xaa m  is Met or Ala; and Xaa 70  is a sequence of 70 amino acids.    
     
     
         10 . An isolated polypeptide encoded by DNA selected from the group consisting of: 
 a) DNA capable of hybridizing under highly stringent conditions to a probe consisting essentially of nucleotides 310 to 1684 of SEQ ID NO:1; and    b) DNA capable of hybridizing under highly stringent conditions to DNA complementary to the probe of a), 
 wherein the highly stringent conditions comprise a hybridizing temperature of at least 63° C.  
   
     
     
         11 . A soluble LIR polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of: 
 a) the extracellular domain of LIR family members, the extracellular domains selected from the group consisting of 
 amino acids x 1  to 458 of SEQ ID NO:2, wherein x 1  is amino acid 1 or 17;  
 amino acids x 2  to 458 of SEQ ID NO:4, wherein x 2  is amino acid 1 or 17;  
 amino acids x 3  to 439 of SEQ ID NO:8, wherein x 3  is amino acid 1 or 17;  
 amino acids x 4  to 458 of SEQ ID NO:10, wherein x 4  is amino acid 1 or 17,  
 amino acids x 5  to 242 of SEQ ID NO:12, wherein x 5  is amino acid 1 or 17;  
 amino acids x 6  to 461 of SEQ ID NO:14, wherein x 6  is amino acid 1 or 17;  
 amino acids x 7  to 449 of SEQ ID NO:16, wherein x 7  is amino acid 1 or 17;  
 amino acids x 8  to 259 of SEQ ID NO:18, wherein x 8  is amino acid 1 or 17;  
 amino acids x 9  to 443 of SEQ ID NO:20, wherein x 9  is amino acid 1 or 17; and,  
 amino acids x 10  to 456 of SEQ ID NO:22, wherein x 10  is amino acid 1 or 17;  
   b) a fragment of any of the LIR extracellular domains of a), wherein the fragment is capable of binding a ligand.    
     
     
         12 . A soluble polypeptide of  claim 11  comprising an amino acid sequence selected from the group consisting of: 
 a) the extracellular domain of LIR family members, the extracellular domains selected from the group consisting of 
 amino acids x 1  to 458 of SEQ ID NO:2, wherein x 1  is amino acid 1 or 17;  
 amino acids x 2  to 458 of SEQ ID NO:4, wherein x 2  is amino acid 1 or 17;  
 amino acids x 3  to 439 of SEQ ID NO:8, wherein x 3  is amino acid 1 or 17;  
 amino acids x 4  to 458 of SEQ ID NO:10, wherein x 4  is amino acid 1 or 17;  
 amino acids x 5  to 242 of SEQ ID NO:12, wherein x 5  is amino acid 1 or 17;  
 amino acids x 6  to 461 of SEQ ID NO:14, wherein x 6  is amino acid 1 or 17;  
 amino acids x 7  to 449 of SEQ ID NO:16, wherein x 7  is amino acid 1 or 17;  
 amino acids x 8  to 259 of SEQ ID NO:18, wherein x 8  is amino acid 1 or 17;  
 amino acids x 9  to 443 of SEQ ID NO:20, wherein x 9  is amino acid 1 or 17; and,  
 amino acids x 10  to 456 of SEQ ID NO:22, wherein x 10  is amino acid 1 or 17;  
 
 b) a fragment of any of the human P3G2 extracellular domains of a).  
 
     
     
         13 . An isolated polypeptide encoded by a DNA selected from the group consisting of: 
 a) DNA capable of hybridizing under highly stringent conditions to DNA selected from the group consisting of SEQ ID NO:5 and SEQ ID NO:6, the highly stringent conditions including a hybridizing temperature of at least 68° C.    
     
     
         14 . A fusion protein comprising amino acids 17 to 458 of SEQ ID NO:2 and the Fc region of Ig.  
     
     
         15 . A fusion DNA construct comprising DNA encoding amino acids 17 to 458 of SEQ ID NO:2 and DNA encoding the Fc region of Ig.  
     
     
         16 . A recombinant expression vector comprising DNA of  claim 1 .  
     
     
         17 . A process for preparing an LIR polypeptide, the process comprising culturing a host cell transformed with an expression vector of  claim 16  under conditions that promote expression of said polypeptide, and recovering said polypeptide.  
     
     
         18 . A composition comprising a suitable diluent carrier and a polypeptide of  claim 8 .  
     
     
         19 . A host cell transformed or transfected with an expression vector according to  claim 16 .  
     
     
         20 . An antibody that is immunoreactive with a polypeptide of  claim 8.