Novel co-stimulatory molecules

Abstract

The invention provides polynucleotides and polypeptides encoded therefrom having advantageous properties, including an ability of the polypeptides to preferentially bind a CD28 or CTLA-4 receptor at a level greater or less than the ability of human B7-1 to bind CD28 or CTLA-4, or to induce or inhibit altered level of T cell proliferation response greater compared to that generated by human B7-1. The polypeptides and polynucleotides of the invention are useful in therapeutic and prophylactic treatment methods, gene therapy applications, and vaccines.

1 . An isolated or recombinant polypeptide comprising an amino acid sequence of an extracellular domain, wherein said extracellular domain amino acid sequence has at least 90% amino acid sequence identity to an extracellular domain amino acid sequence of at least one of SEQ ID NOS:48-68, 174-221, 283-285, and 290-293, and is not a naturally-occurring extracellular domain amino acid sequence, and wherein said polypeptide has a CD28/CTLA-4 binding affinity ratio equal to or greater than the CD28/CTLA-4 binding affinity ratio of human B7-1, or has an ability to induce a T-cell proliferative response equal to or greater than that of human B7-1. 
     
     
         2 .- 32 . (canceled) 
     
     
         33 . The polypeptide of  claim 1 , wherein the polypeptide comprises a fusion protein comprising at least one additional amino acid sequence. 
     
     
         34 . The polypeptide of  claim 33 , wherein the at least one additional amino acid sequence comprises at least one Ig polypeptide. 
     
     
         35 .- 42 . (canceled) 
     
     
         43 . An isolated or recombinant nucleic acid comprising a polynucleotide sequence
 which encodes a polypeptide comprising an amino acid sequence of an extracellular domain, wherein said extracellular domain amino acid sequence has at least 90% amino acid sequence identity to an extracellular domain amino acid sequence of at least one of SEQ ID NOS:48-68, 174-221, 283-285, and 290-293, and is not a naturally-occurring extracellular domain amino acid sequence, and wherein said polypeptide has a CD28/CTLA-4 binding affinity ratio equal to or greater than the CD28/CTLA-4 binding affinity ratio of human B7-1 or has an ability to induce a T cell proliferation response that is equal to or greater than that induced by human B7-1.   
     
     
         44 .- 57 . (canceled) 
     
     
         58 . A cell or vector comprising the nucleic acid of  claim 43 . 
     
     
         59 .- 79 . (canceled) 
     
     
         80 . An isolated or recombinant polypeptide comprising:
 (1) an amino acid sequence having at least 95% identity to at least one of SEQ ID NOS:69-92, 222-252, 286-289, or a subsequence thereof comprising an extracellular domain, wherein said amino acid sequence (a) is a non naturally-occurring amino acid sequence, and (b) comprises at least one of:   Gly at position 2; Thr at position 4; Arg at position 5; Gly at position 8; Pro at position 12; Met at position 25; Cys at position 27; Pro at position 29; Leu at position 31; Arg at position 40; Leu at position 52; His at position 65; Ser at position 78; Asp at position 80; Tyr at position 87; Lys at position 120; Asp at position 122; Lys at position 129; Met at position 135; Phe at position 150; Ile at position 160; Ala at position 164; His at position 172; Phe at position 174; Leu at position 176; Asn at position 178; Asn at position 186; Glu at position 194; Gly at position 196; Thr at position 199; Ala at position 210; His at position 212; Arg at position 219; Pro at position 234; Asn at position 241; Leu at position 244; Thr at position 250; Ala at position 254; Tyr at position 265; Arg at position 266; Glu at position 273; Lys at position 275; Ser at position 276; an amino acid deletion at position 276; or Thr at position 279, wherein the position number corresponds to that of the human B7-1 amino acid sequence (SEQ ID NO:278),   (2) an amino acid sequence that differs from a primate B7-1 amino acid sequence in at least one mutation selected from: Ser 12 Pro; Leu 25 Met; Gly 27 Cys; Ser 29 Pro; Lys 40 Arg; His 52 Leu; Tyr 65 His; Glu 122 Asp; Glu 129 Lys; Thr 135 Met; Thr 164 Ala; Ser 174 Phe; Glu 196 Gly; Ala 199 Thr; Thr 210 Ala; Lys 219 Arg; Thr 234 Pro; Asp 241 Asn; Val 254 Ala; Val 254 Ala; Arg 275 Lys; Arg 276 Ser; or Arg 279 Thr; the mutation being indicated comprising a mutation relative to human B7-1 with the amino acid sequence shown in SEQ ID NO:278, or   (3) an amino acid sequence, said amino acid sequence having at least 90% identity to at least one polypeptide sequence of SEQ ID NOS:263-272, or a subsequence thereof comprising the extracellular domain, wherein said amino acid sequence is not a naturally-occurring amino acid sequence,   wherein said polypeptide has a CTLA-4/CD28 binding affinity ratio equal to or greater than the CTLA-4/CD28 binding affinity ratio of human B7-1 or an ability to induce a T cell proliferation response that is equal to or less than that of human B7-1.   
     
     
         81 .- 117 . (canceled) 
     
     
         118 . The polypeptide of  claim 80 ,  97 ,  101 ,  102 ,  107 , or  113 , wherein the polypeptide comprises a fusion protein comprising at least one additional amino acid sequence. 
     
     
         119 . The polypeptide of  claim 118 , wherein the at least one additional amino acid sequence comprises at least one Ig polypeptide. 
     
     
         120 .- 127 . (canceled) 
     
     
         128 . An isolated or recombinant nucleic acid comprising a polynucleotide sequence which
 (1) encodes a polypeptide comprising an amino acid sequence haying., at least 95% identity to at least one of SEQ ID NOS:69-92, 222-252, 286-289, or a subsequence thereof comprising an extracellular domain, wherein said amino acid sequence (a) is a non naturally-occurring amino acid sequence, and (b) comprises at least one of:   Gly at position 2; Thr at position 4; Arg at position 5; Gly at position 8; Pro at position 12; Met at position 25; Cys at position 27; Pro at position 29; Leu at position 31; Arg at position 40; Leu at position 52; His at position 65; Ser at position 78; Asp at position 80; Tyr at position 87; Lys at position 120; Asp at position 122; Lys at position 129; Met at position 135; Phe at position 150; Ile at position 160; Ala at position 164; His at position 172; Phe at position 174; Leu at position 176; Asn at position 178; Asn at position 186; Glu at position 194; Gly at position 196; Thr at position 199; Ala at position 210; His at position 212; Arg at position 219; Pro at position 234; Asn at position 241; Leu at position 244; Thr at position 250; Ala at position 254; Tyr at position 265; Arg at position 266; Glu at position 273; Lys at position 275; Ser at position 276; an amino acid deletion at position 276; and Thr at position 279, wherein the number of the amino acid position corresponds to that of the human B7-1 amino acid sequence (SEQ ID NO:278), or   (2) encodes a polypeptide comprising an amino acid sequence, said amino acid sequence having at least 75% identity to at least one polypeptide sequence of SEQ ID NOS:263-272, or a subsequence thereof comprising the extracellular domain, wherein said amino acid sequence is a non naturally-occurring sequence,   and wherein said polypeptide has a CTLA-4/CD28 binding affinity ratio about equal to or greater than the CTLA-4/CD28 binding affinity ratio of human B7-1 and/or an ability to induce a T cell proliferation response that is equal to or greater than that induced by human B7-1.   
     
     
         129 .- 173 . (canceled) 
     
     
         174 . A method of producing a polypeptide, the method comprising:
 (a) introducing into a population of cells a nucleic acid of  claim 43 , the nucleic acid operatively linked to a regulatory sequence effective to produce the encoded polypeptide;   (b) culturing the cells in a culture medium to produce the polypeptide; and   (c) isolating the polypeptide from the cells or from the culture medium.   
     
     
         175 .- 178 . (canceled) 
     
     
         179 . A method of modifying T-cell proliferation, the method comprising: contacting a population of T cells with a polypeptide of  claim 1 , thereby modifying proliferation of the T cells. 
     
     
         180 .- 181 . (canceled) 
     
     
         182 . A method of treating an autoimmune disorder or medical condition in a subject, the method comprising: administering to the subject an effective amount of the polypeptide of  claim 1 . 
     
     
         183 .- 320 . (canceled) 
     
     
         321 . A cell or vector comprising the nucleic acid of  claim 128 . 
     
     
         322 . A method of producing a polypeptide, the method comprising:
 (a) introducing into a population of cells a nucleic acid of  claim 128 , the nucleic acid operatively linked to a regulatory sequence effective to produce the encoded polypeptide;   (b) culturing the cells in a culture medium to produce the polypeptide; and   (c) isolating the polypeptide from the cells or from the culture medium.   
     
     
         323 . A method of modifying T-cell proliferation, the method comprising: contacting a population of T cells with a polypeptide of  claim 80 , thereby modifying proliferation of the T cells. 
     
     
         324 . A method of treating an autoimmune disorder or medical condition in a subject, the method comprising: administering to the subject an effective amount of the polypeptide of  claim 80 .