Co-stimulatory polypeptides
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 extracellular domain, said extracellular domain comprising an amino acid sequence having at least 91% sequence identity to a subsequence of the polypeptide sequence set forth in SEQ ID NO:66, wherein the subsequence is the extracellular domain of SEQ ID NO:66, and wherein the isolated or recombinant polypeptide has a human CD28/human CTLA-4 binding affinity ratio equal to or greater than the human CD28/human CTLA-4 binding affinity ratio of human B7-1 when said isolated or recombinant polypeptide is expressed on a cell or bound to a cell membrane.
2. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide is expressed on a cell or bound to a cell membrane.
3. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide comprises an extracellular domain which comprises an amino acid sequence having at least 95% sequence identity to the extracellular domain of SEQ ID NO:66, wherein said extracellular domain of SEQ ID NO:66 comprises at least amino acid residues 35–244 of SEQ ID NO:66.
4. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide comprises an extracellular domain which comprises an amino acid sequence having at least 95% sequence identity to the extracellular domain of SEQ ID NO:66, wherein the extracellular domain of SEQ ID NO;66 comprises at least amino acid residues 35–245 of SEQ ID NO:66.
5. The isolated or recombinant polypeptide of claim 3 , wherein the polypeptide comprises art extracellular domain comprising at least amino acid residues 35–244 of SEQ ID NO:66.
6. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide further comprises a signal peptide.
7. The isolated or recombinant polypeptide of claim 6 , wherein the signal peptide comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence comprising residues 1–34 of SEQ ID NO:66.
8. The isolated or recombinant polypeptide of claim 6 , wherein the signal peptide has an amino acid sequence comprising amino acid residues 1–34 of SEQ ID NO:66.
9. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide comprises a transmembrane domain.
10. The isolated or recombinant polypeptide of claim 9 , wherein the polypeptide comprises the transmembrane domain of SEQ ID NO:66.
11. The isolated or recombinant polypeptide of claim 10 , wherein the transmembrane domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence comprising at least amino acid residues 245–268 or 246–272 of SEQ ID NO:66.
12. The isolated or recombinant polypeptide of claim 11 , wherein the transmembrane domain comprises an amino acid sequence comprising at least amino acid residues 245–268 or 246–272 of SEQ ID NO:66.
13. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide further comprises a cytoplasmic domain.
14. The isolated or recombinant polypeptide of claim 13 , wherein the polypeptide comprises the cytoplasmic domain of SEQ ID NO:66.
15. The isolated or recombinant polypeptide of claim 13 , wherein the cytoplasmic domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence comprising at least amino acid residues 269–303 or 273–303 of SEQ ID NO:66.
16. The isolated or recombinant polypeptide of claim 15 , wherein the cytoplasmic domain comprises an amino acid sequence comprising at least amino acid residues 269–303 or 273–303 of SEQ ID NO:66.
17. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence comprising a mature domain of SEQ ID NO:66.
18. The isolated or recombinant polypeptide of claim 17 , wherein the mature domain comprises amino acid residues 35–303 of SEQ ID NO:66.
19. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence corresponding to the signal peptide, extracellular domain and transmembrane domain of SEQ ID NO:66 which comprises at least amino acid residues 1–268 or 1–272 of SEQ ID NO:66.
20. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide has at least 91% sequence identity to the full length amino acid sequence of SEQ ID NO:66.
21. The isolated or recombinant polypeptide of claim 20 , wherein the polypeptide comprises the full length amino acid sequence of SEQ ID NO:66.
22. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide has a human CD28/human CTLA-4 binding affinity ratio greater than the human CD28/human CTLA-4 binding affinity ratio of human B7-1.
23. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide has a binding affinity for CD28 that is greater than the binding affinity of human B7-1 for CD28.
24. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide has a binding affinity for CTLA-4 that is less than the binding affinity of human B7-1 for CTLA-4.
25. The isolated or recombinant polypeptide of claim 1 , wherein the polypeptide has an ability to induce a T-cell proliferation response equal to or greater than the T-cell proliferation response induced by human B7-1.
26. An isolated or recombinant polypeptide comprising an extracellular domain, said extracellular domain comprising an amino acid sequence having at least 91% sequence identity to a subsequence of SEQ ID NO:66, said subsequence comprising at least amino acid residues 35–244 or 35–245 of SEQ ID NO:66, wherein said polypeptide induces a T-cell proliferation response equal to or greater than the T-cell proliferation response induced by human B7-1 when said polypeptide is expressed on a cell or bound to a cell membrane.
27. The isolated or recombinant polypeptide of claim 26 , wherein the extracellular domain of said polypeptide comprises an amino acid sequence having at least 95% sequence identity to the subsequence of SEQ ID NO:66 that comprises at least amino acid residues 35–244.
28. The isolated or recombinant polypeptide of claim 26 , wherein the polypeptide comprises one or more of a signal peptide, transmembrane domain, and cytoplasmic domain.
29. The isolated or recombinant polypeptide of claim 28 , wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to the full length amino acid sequence of SEQ ID NO:66.
30. The isolated or recombinant polypeptide of claim 1 , wherein the cell is an antigen-presenting cell.
31. The polypeptide of claim 1 , comprising at least one modified amino acid.
32. The polypeptide of claim 30 , wherein the modified amino acid is selected from: a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, an amino acid conjugated to a lipid moiety, and an amino acid conjugated to an organic derivariting agent.
33. An isolated or recombinant polypeptide comprising an amino acid sequence according to the formula:
MGHTM-X6-W-X8-SLPPK-X14-PCL-X18-X19-X20-QLLVLT-X27-LFYFCSGITPKSVTKRVKETVMLSCDY-X55-TSTE-X60-LTSLRIYW-X69-KDSKMVLAILPGKVQVWPEYKNRTITDMNDN-X101-RIVI-X106-ALR-X110-SD-X113-GTYTCV-X120-QKP-X124-LKGAYKLEHL-X135-SVRLMIRADFPVP-X149-X150-X151-DLGNPSPNIRRLICS-X167-X168-X169-GFPRPHL-X177-WLENGEELNATNTT-X192-SQDP-X197-T-X199-LYMISSEL-X208-FNVTNN-X215-SI-X218-CLIKYGEL-X227-VSQIFPWSKPKQEPPIDQLPF-X249-VIIPVSGALVL-X261-A-X263-VLY-X267-X268-ACRH-X273-ARWKRTRRNEETVGTE RLSPIYLGSAQSSG (SEQ ID NO:284), or a extracellullar domain subsequence thereof comprising amino acid residues at positions 35–244,
wherein the amino acid residue at position X6 is Lys or Glu; position X8 is Arg or Gly; position X14 is Arg or Cys; position X18 is Trp or Arg; position X19 is Pro or Leu; position X20 is Ser or Pro; position X27 is Asp or Gly; position X55 is Asn or Ser; position X60 is Glu or Lys; position X69 is Gln or Mg; position X101 is Pro or Leu; position X106 is Leu or Gln; position X110 is Pro or Leu; position X113 is Lys or Ser; position 120 is Val or Ile; position X124 is Val or Asp; position X135 is Thr or Ala; position X149 is Thr, Ser, or deleted; position X150 is Ile or deleted; position X151 is Asn or Thr; position X167 is Thr or deleted; position X169 is Ser or deleted; position X169 is Gly or deleted; position X117 is Cys or Tyr; position X192 is Val or Leu; position X197 is Gly or Glu; position X199 is Glu or Lys; position X208 is Gly or Asp; position X215 is His or Arg; position X218 is Ala or Val; position X227 is Ser or Leu; position X249 is Trp, Leu, or Arg; position X261 is Ala or Thr; position X263 is Val Ala, or Ile; position X267 is Arg or Cys; position X268 is Pro or Leu; and position X273 is Gly or Val, and
wherein the polypeptide has a human CD28/human CTLA-4 binding affinity ratio equal to or greater than the human CD28/human CTLA-4 binding affinity ratio of human B7-1 and/or induces a T-cell proliferation or activation response when the isolated or recombinant polypeptide is expressed on a cell or bound to a cell membrane.
34. The isolated or recombinant polypeptide of claim 33 , wherein the polypeptide has a human CD28/human CTLA-4 binding affinity ratio greater than the human CD28/human CTLA-4 binding affinity ratio of human B7-1.
35. The isolated or recombinant polypeptide of claim 33 , wherein the polypeptide induces a T-cell proliferation response equal to, or greater than that induced by human B7-1.
36. The isolated or recombinant polypeptide of claim 33 , comprising three or more of: Lys at position X6; Arg at position X8; Arg at position X14; Trp at position X18; Pro at position X19; Ser at position X20; Asp at position X27; Asn at position X55; Len at position X106; Pro at position X110; Lys at position X113; Val at position X120; Val at position X124; Thr at position X135; Asn at position X151; Cys at position X177; Val at position X192; Gly at position X197; Glu at position X199; Gly at position X208; His at position 215; Ala at position X218; Trp at position X249; Ala at position X261; Val at position X263; Arg at position X267; Pro at position X268; and Gly at position X273.
37. The isolated or recombinant polypeptide of claim 36 , comprising three or more of: Arg at position X8; Arg at position X14; Trp at position X18; Pro at position X19; Ser at position X20; Pro at position X110; Val at position X120; Val at position X124; Cys at position X177; Val at position X192; Gly at position X197; Glu at position X199; Gly at position X208; His at position X215; Ala at position X218; Trp at position X249; Ala at position X261; and Val at position X263.
38. The isolated or recombinant polypeptide of claim 37 , comprising amino acid residues 35–244 of SEQ ID NO:66.
39. An isolated or recombinant polypeptide comprising an amino acid sequence having at least 91% sequence identity to the complete amino acid sequence set forth in SEQ ID NO:66, wherein said polypeptide when expressed on a cell or bound to a cell membrane has a human CD28/human CTLA-4 binding affinity ratio at least equal to the human CD28/human CTLA-4 binding affinity ratio of human B7-1 or induces a T-cell proliferation or activation response.
40. A phamaceutical composition comprising a polypeptide of claim 1 and a pharmaceutically acceptable excipient.
41. An isolated or recombinant polypeptide comprising an amino acid sequence comprising at least amino acid residues 245–268 or 246–272 of SEQ ID NO:66.
42. An isolated or recombinant polypeptide comprising an amino acid sequence comprising at least amino acid residues 269–303 or 273–303 of SEQ ID NO:66.
43. The isolated or recombinant polypeptide of claim 6 , wherein the polypeptide comprises an amino acid sequence comprising at least amino acid residues 1–244 or 1–245 of SEQ ID NO:66.
44. The isolated or recombinant polypeptide of claim 27 , wherein said extracellular domain comprises an amino acid sequence having at least 97% sequence identity to the subsequence of SEQ ID NO:66 comprising at least amino acid residues 35–244.
45. The isolated or recombinant polypeptide of claim 27 , wherein said polypeptide when expressed on a cell or bound to a cell membrane induces a T-cell proliferation response greater than the T-cell proliferation response induced by human B7-1.
46. The isolated or recombinant polypeptide of claim 39 , wherein said polypeptide comprises an amino acid sequence having at least 95% sequence identity to the complete amino acid sequence of SEQ ID NO:66, and wherein said polypeptide when expressed on a cell or bound to a cell membrane has a human CD28/human CTLA-4 binding affinity ratio that is at least equal to the human CD28/CTLA-4 binding affinity ratio of human B7-1.
47. The isolated or recombinant polypeptide of claim 46 , wherein said polypeptide when expressed on a cell or bound to a cell membrane has a human CD28/human CTLA-4 binding affinity ratio that is greater than the human CD28/CTLA-4 binding affinity ratio of human B7-1.
48. The isolated or recombinant polypeptide of claim 39 , wherein said polypeptide comprises an amino acid sequence having at least 95% sequence identity to the complete amino acid sequence of SEQ ID NO:66, and wherein said polypeptide when expressed on a cell or bound to a cell membrane induces a T-cell proliferation or activation response.
49. The isolated or recombinant polypeptide of claim 48 , wherein said polypeptide comprises an amino acid sequence having at leas 95% sequence identity to the complete amino acid sequence of SEQ ID NO:66, and wherein said polypeptide when expressed on a cell or bound to a cell membrane induces a T-cell proliferation or activation response.
50. The isolated or recombinant polypeptide of claim 49 , wherein said polypeptide when expressed on a cell or bound to a cell membrane induces a T-cell proliferation or activation response greater than that induced by human B7-1.
51. A pharmaceutical composition comprising a polypeptide of claim 39 and a pharmaceutically acceptable excipient.
52. A pharmaceutical composition comprising a polypeptide of claim 44 and a pharmaceutically acceptable excipient.
53. A pharmaceutical composition comprising a polypeptide of claim 46 and a pharmaceutically acceptable excipient.