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.

1 - 20  (canceled)  
     
     
         21 : An isolated polypeptide comprising at least 46 contiguous amino acids having a sequence identity score of 77 to 100% to residues 5 to 50 as shown in SEQ ID NO: 2.  
     
     
         22 : The isolated polypeptide as in  claim 21  which comprises the extracellular domain of LIR-pbm8 (SEQ ID NO: 10).  
     
     
         23 : The isolated polypeptide as in  claim 22 , wherein said extracellular domain excludes the signal peptides.  
     
     
         24 : The isolated polypeptide as in  claim 21  which comprises the amino acid sequence of LIR-pbm25 (SEQ ID NO: 8).  
     
     
         25 : The isolated polypeptide as in  claim 24 , wherein said amino acid sequence excludes the signal peptides.  
     
     
         26 : The isolated polypeptide as in  claim 21  which comprises the extracellular domain of LIR-P3G2 (SEQ ID NO: 2) or the extracellular domain of LIR-18A3 (SEQ ID NO: 4).  
     
     
         27 : The isolated polypeptide as in  claim 26 , wherein said extracellular domain excludes the signal peptides.  
     
     
         28 : The isolated polypeptide as in  claim 21  which further comprises an amino acid sequence having a sequence identity score of at least 80% to the cytoplasmic domain of a LIR member selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         29 : The isolated polypeptide as in  claim 28 , wherein the sequence identity score to the cytoplasmic domain is at least 90%.  
     
     
         30 : The isolated polypeptide as in  claim 28 , wherein the sequence identity score to the cytoplasmic domain is 100%.  
     
     
         31 : An isolated polypeptide which is soluble, said polypeptide comprising an amino acid sequence selected from the group consisting of: 
 (a) an amino acid sequence comprising at least 46 contiguous amino acids that are 77 to 100% identical to residues 5 to 50 of SEQ ID NO: 2;    (b) an amino acid sequence comprising at least 46 contiguous amino acids that are 90% identical to residues 5 to 50 of SEQ ID NO: 2;    (c) an amino acid sequence comprising at least 46 contiguous amino acids that are 96% identical to residues 5 to 50 of SEQ ID NO: 2;    (d) an amino acid sequence comprising residues 5 to 50 of SEQ ID NO: 2;    (e) an amino acid sequence that is at least 80% identical to the extracellular domain of LIR-P3G2 (SEQ ID NO: 2), wherein said extracellular domain includes or excludes its signal peptides;    (f) an amino acid sequence that is at least 90% identical to the extracellular domain of LIR-P3G2 (SEQ ID NO: 2), wherein said extracellular domain includes or excludes its signal peptides;    (g) an amino acid sequence that comprises the extracellular domain of LIR-P3G2 (SEQ ID NO: 2), wherein said extracellular domain includes or excludes its signal peptides;    (h) an amino acid sequence that is at least 80% identical to the extracellular domain of LIR-18A3 (SEQ ID NO: 4), wherein said extracellular domain includes or excludes its signal peptides;    (i) an amino acid sequence that is at least 90% identical to the extracellular domain of LIR-18A3 (SEQ ID NO: 4), wherein said extracellular domain includes or excludes its signal peptides;    (j) an amino acid sequence that comprises the extracellular domain of LIR-18A3 (SEQ ID NO: 4), wherein said extracellular domain includes or excludes its signal peptides;    (k) an amino acid sequence that is at least 80% identical to the extracellular domain of LIR-pbm8 (SEQ ID NO: 10), wherein said extracellular domain includes or excludes its signal peptides;    (l) an amino acid sequence that is at least 90% identical to the extracellular domain of LIR-pbm8 (SEQ ID NO: 10), wherein said extracellular domain includes or excludes its signal peptides;    (m) an amino acid sequence that comprises the extracellular domain of LIR-pbm8 (SEQ ID NO: 10), wherein said extracellular domain includes or excludes its signal peptides;    (n) an amino acid sequence that is at least 80% identical to LIR-pbm25 (SEQ ID NO: 8), wherein said LIR-pbm25 includes or excludes its signal peptides;    (o) an amino acid sequence that is at least 90% identical to LIR-pbm25 (SEQ ID NO: 8), wherein said LIR-pbm25 includes or excludes its signal peptides; and    (p) an amino acid sequence that comprises LIR-pbm25 (SEQ ID NO: 8), wherein said LIR-pbm25 includes or excludes its signal peptides.    
     
     
         32 : A fusion protein comprising the polypeptide as in  claim 31 .  
     
     
         33 : The fusion protein as in  claim 32  comprising a peptide moiety capable of promoting oligomerization of said fusion protein.  
     
     
         34 : The fusion protein as in  claim 33 , wherein said peptide moiety comprises a leucine zipper domain or a Fc polypeptide.  
     
     
         35 : The fusion protein as in  claim 34 , wherein said peptide moiety comprises a Fc polypeptide.  
     
     
         36 : The fusion protein as in  claim 35 , wherein said Fc polypeptide is either a native or a mutein thereof.  
     
     
         37 : The fusion protein as in  claim 32  comprising an antigenic identification polypeptide.  
     
     
         38 : The fusion protein as in  claim 37 , wherein said antigenic identification polypeptide comprises a poly-His polypeptide or a FLAG polypeptide.  
     
     
         39 : An oligomer comprising at least two monomers, said monomer comprising the polypeptide as in  claim 31 .  
     
     
         40 : The oligomer as in  claim 39  which comprises at least three monomers.  
     
     
         41 : An oligomer comprising at least two monomers, said monomer comprising the fusion protein as in  claim 32 .  
     
     
         42 : The oligomer as in  claim 41  which comprises at least three monomers.  
     
     
         43 : A composition comprising the polypeptide as in  claim 31 , in a pharmaceutically acceptable carrier or diluent.  
     
     
         44 : A composition comprising the fusion protein as in  claim 32 , in a pharmaceutically acceptable carrier or diluent.  
     
     
         45 : A composition comprising the oligomer as in  claim 39 , in a pharmaceutically acceptable carrier or diluent.  
     
     
         46 : A composition comprising the oligomer as in claims  41 , in a pharmaceutically acceptable carrier or diluent.  
     
     
         47 : An antibody which specifically binds to a polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4), LIR-pbm25 (SEQ ID NO: 8) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         48 : The antibody as in  claim 47  which is a monoclonal antibody or an immunoglobulin fragment thereof.  
     
     
         49 : A composition comprising the antibody as in  claim 47 , in a pharmaceutically acceptable carrier or diluent.  
     
     
         50 : A method for producing the antibody as in  claim 47 , comprising inoculating an animal with an amino acid sequence of said polypeptide, or an immunogenic fragment thereof, under conditions in which said animal produces an antibody which specifically binds to said polypeptide, and isolating said antibody.  
     
     
         51 : An isolated nucleic acid comprising (a) nucleotides 310 to 1684 of SEQ ID NO: 1, or (b) a nucleotide sequence capable of hybridizing to the complement of (a) under high stringency condition, said nucleic acid encoding a polypeptide comprising at least 46 contiguous amino acids which are 77 to 100% identical to amino acids 5 to 50 of SEQ ID NO: 2.  
     
     
         52 : The isolated nucleic acid as in  claim 51 , wherein said polypeptide is at least 80% identical to a polypeptide selected from the group consisting of: (a) the extracellular domain of LIR-P3G2 (SEQ ID NO: 2), (b) the extracellular domain of LIR-18A3 (SEQ ID NO: 4), (c) the extracellular domain of LIR-pbm8 (SEQ ID NO: 10), (d) LIR-pbm25 (SEQ ID NO: 8) and (e) any of (a) to (d) from which the signal peptides are excluded.  
     
     
         53 : The isolated nucleic acid as in  claim 51 , wherein said polypeptide is at least 90% identical to a polypeptide selected from the group consisting of: (a) the extracellular domain of LIR-P3G2 (SEQ ID NO: 2), (b) the extracellular domain of LIR-18A3 (SEQ ID NO: 4), (c) the extracellular domain of LIR-pbm8 (SEQ ID NO: 10), (d) LIR-pbm25 (SEQ ID NO: 8) and (e) any of (a) to (d) from which the signal peptides are excluded.  
     
     
         54 : The isolated nucleic acid as in  claim 51 , wherein said polypeptide is selected from the group consisting of: (a) the extracellular domain of LIR-P3G2 (SEQ ID NO: 2), (b) the extracellular domain of LIR-18A3 (SEQ ID NO: 4), (c) the extracellular domain of LIR-pbm8 (SEQ ID NO: 10), (d) LIR-pbm25 (SEQ ID NO: 8) and (e) any of (a) to (d) from which the signal peptides are excluded.  
     
     
         55 : The isolated nucleic acid as in  claim 52 , wherein said nucleic acid further comprises a nucleotide sequence which encodes an amino acid sequence which is at least 80% identical to the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         56 : The isolated nucleic acid as in  claim 52 , wherein said nucleic acid further comprises a nucleotide sequence which encodes an amino acid sequence which is at least 90% identical to the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         57 : The isolated nucleic acid as in  claim 52 , wherein said nucleic acid further comprises a nucleotide sequence which encodes the amino acid sequence of the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         58 : The isolated nucleic acid as in  claim 53 , wherein said nucleic acid further comprises a nucleotide sequence which encodes an amino acid sequence which is at least 80% identical to the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         59 : The isolated nucleic acid as in  claim 53 , wherein said nucleic acid further comprises a nucleotide sequence which encodes an amino acid sequence which is at least 90% identical to the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         60 : The isolated nucleic acid as in  claim 53 , wherein said nucleic acid further comprises a nucleotide sequence which encodes the amino acid sequence of the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         61 : The isolated nucleic acid as in  claim 54 , wherein said nucleic acid further comprises a nucleotide sequence which encodes an amino acid sequence which is at least 80% identical to the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         62 : The isolated nucleic acid as in  claim 54 , wherein said nucleic acid further comprises a nucleotide sequence which encodes an amino acid sequence which is at least 90% identical to the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         63 : The isolated nucleic acid as in  claim 54 , wherein said nucleic acid further comprises a nucleotide sequence which encodes the amino acid sequence of the cytoplasmic domain of a LIR polypeptide selected from the group consisting of: LIR-P3G2 (SEQ ID NO: 2), LIR-18A3 (SEQ ID NO: 4) and LIR-pbm8 (SEQ ID NO: 10).  
     
     
         64 : The isolated nucleic acid of  claim 51  which comprises (a) SEQ ID NO: 1, 3, 7, or 9, or (b) a nucleotide sequence capable of hybridizing under high stringency condition to the complement of SEQ ID NO: 1, 3, 7, or 9.  
     
     
         65 : A vector comprising the nucleic acid as in  claim 51 .  
     
     
         66 : The vector as in  claim 65 , wherein the vector is an expression vector.  
     
     
         67 : A cell comprising the vector as in  claim 65 .  
     
     
         68 : A cell comprising the expression vector as in  claim 66 .  
     
     
         69 : A process for preparing a LIR polypeptide, said process comprising culturing the cell as in  claim 68  under conditions that promote expression of said polypeptide, and recovering said polypeptide.  
     
     
         70 : An oligonucleotide for detecting or preparing a nucleic acid encoding a LIR polypeptide, said oligonucleotide comprises at least 17 contiguous nucleotides and is capable of hybridizing under high stringency condition to SEQ ID NO: 1 or to its complement, wherein said LIR polypeptide comprises at least 46 contiguous amino acids that are 77 to 100% identical to amino acids 5 to 50 of SEQ ID NO: 2.  
     
     
         71 : The oligonucleotide as in  claim 70  which comprises at least 30 contiguous nucleotides.  
     
     
         72 : The oligonucleotide as in  claim 70 , wherein said portion comprises at least 17 contiguous nucleotides which are located between nucleotides 358 and 1684 of said SEQ ID NO: 1.  
     
     
         73 : The oligonucleotide as in  claim 70 , wherein said portion comprises at least 17 contiguous nucleotides which are located between nucleotides 322 and 459 of said SEQ ID NO: 1.  
     
     
         74 : The oligonucleotide as in  claim 70 , wherein said oligonucleotide comprises at least 17 contiguous nucleotides and is capable of hybridizing under high stringency condition to a sequence selected from the group consisting of: SEQ ID NOS: 1, 3, 7 and 9.  
     
     
         75  A method of preparing a nucleic acid encoding a LIR polypeptide, comprising hybridizing a nucleic acid with the oligonucleotide as in  claim 51  under high stringency condition, and isolating the nucleic acid that hybridizes to said oligonucleotide under said condition, wherein said LIR polypeptide comprises at least 46 contiguous amino acids that are 77 to 100% identical to amino acids 5 to 50 of SEQ ID NO: 2.  
     
     
         76 : An oligonucleotide of at least 14 nucleotides in length targeted to a nucleic acid encoding a LIR polypeptide which comprises at least 46 contiguous amino acids that are 77 to 100% identical to amino acids 5 to 50 of SEQ ID NO: 2, said oligonucleotide is capable of hybridizing under high stringency condition to SEQ ID NO: 1, or to its complement, and which is capable of blocking expression of said LIR polypeptide.  
     
     
         77 : The oligonucleotide as in  claim 76 , wherein said oligonucleotide comprises a sugar-phosphodiester backbone that is modified.