Nucleic acid molecules encoding B7-DC variants

• US 8,273,864 B2
• Filed: 09/09/2011
• Issued: 09/25/2012
• Est. Priority Date: 10/04/2002
• Status: Active Grant

• Current Assignee: Mayo Foundation For Medical Education And Research (Mayo Clinic)
• Original Assignee: Mayo Foundation For Medical Education And Research (Mayo Clinic)
• Inventors: Chen, Lieping
• Primary Examiner(s): OUSPENSKI, ILIA I
• Application Number:
  US13/228,893
• Publication Number:
  US 20120003220A1
• Time in Patent Office: 382 Days
• Field of Search: None
• US Class Current: 536/23.1
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/70532 B7 molecules, e.g. CD80, CD86

Abstract

Variant costimulatory polypeptides, nucleic acids encoding such polypeptides, and methods for using the polypeptides and nucleic acids to enhance a T cell response are provided herein.

Claims

1. An isolated nucleic acid molecule encoding a variant B7-DC polypeptide comprising an IgV domain, wherein the variant polypeptide is a variant of wild-type B7-DC polypeptide, has altered binding affinity for PD-1 compared to the wild-type B7-DC polypeptide, and comprises a substitution of an amino acid in the A′

, B, C, C′

, C″

, D, E, F, or G β

strand of wild-type B7-DC polypeptide according to SEQ ID NO;

5 or 6.

2. The isolated nucleic acid molecule of claim 1, wherein the variant B7-DC polypeptide is a variant of murine B7-DC.

3. The isolated nucleic acid molecule of claim 1, wherein the variant B7-DC polypeptide is a variant of human B7-DC.

4. The isolated nucleic acid molecule of claim 1, wherein the variant B7-DC polypeptide or fragment thereof has decreased binding affinity for PD-1 compared to wild-type B7-DC polypeptide.

5. The isolated nucleic acid molecule of claim 4, wherein the variant B7-DC polypeptide or fragment thereof enhances an immune response in a mammal.

6. The isolated nucleic acid molecule of claim 1, wherein the variant B7-DC polypeptide or fragment thereof has increased binding affinity for PD-1 compared to wild-type B7-DC polypeptide.

7. The isolated nucleic acid molecule of claim 6, wherein the variant B7-DC polypeptide or fragment thereof enhances an immune response in a mammal.

8. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 111 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

9. The isolated nucleic acid molecule of claim 8, wherein the substitution is with a serine residue.

10. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 113 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

11. The isolated nucleic acid molecule of claim 10, wherein the substitution is with a serine residue.

12. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 56 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

13. The isolated nucleic acid molecule of claim 12, wherein the substitution is with a serine residue.

14. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 67 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

15. The isolated nucleic acid molecule of claim 14, wherein the substitution is with a tyrosine residue.

16. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 71 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

17. The isolated nucleic acid molecule of claim 16, wherein the substitution is with a serine residue.

18. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 101 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

19. The isolated nucleic acid molecule of claim 18, wherein the substitution is with a serine residue.

20. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 105 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

21. The isolated nucleic acid molecule of claim 20, wherein the substitution is with an alanine residue.

22. The isolated nucleic acid molecule of claim 1, wherein the variant polypeptide comprises a substitution of the amino acid at position 58 number from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

23. The isolated nucleic acid molecule of claim 22, wherein the substitution is with a tyrosine residue.

26. A vector comprising the nucleic acid molecule of claim 1.

27. The vector of claim 26, wherein the nucleic acid molecule is operably linked to an expression control sequence.

28. An isolated host cell comprising the vector of claim 27.

29. A method for enhancing an immune response in a mammal comprising administering to the mammal the vector of claim 27.

24. An isolated nucleic acid molecule encoding a polypeptide fragment of murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5), the polypeptide fragment comprising amino acids 67-71, amino acids 101-105, or amino acids 111-113 of SEQ ID NO;

6, wherein the polypeptide inhibits binding of endogenous B7-DC to PD-1.

25. The isolated nucleic acid molecule of claim 24 encoding a polypeptide comprising amino acids 101-113 of murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

30. An isolated nucleic acid molecule encoding a fusion polypeptide comprising a first fusion partner comprising a variant B7-DC polypeptide comprising an IgV domain, wherein the variant polypeptide is a variant of wild-type B7-DC polypeptide, has altered binding affinity for PD-1 compared to the wild-type B7-DC polypeptide, and comprises a substitution of an amino acid in the A′

, B, C, C′

, C″

, D, E, F, or G β

strand of wild-type B7-DC polypeptide according to SEQ ID NO;

5 or 6, and a second fusion partner.

31. The isolated nucleic acid molecule of claim 30, wherein the second fusion partner comprises one or more domains of an Ig heavy chain constant region.

32. The isolated nucleic acid molecule of claim 30, wherein the second polypeptide comprises an amino acid sequence corresponding to the hinge, CH2 and CH3 regions of a human immunoglobulin chain.

33. The isolated nucleic acid molecule of claim 30, wherein the first polypeptide comprises the extracellular domain of B7-DC or a fragment thereof, and wherein the second polypeptide comprises an amino acid sequence corresponding to the hinge, CH2 and CH3 regions of a human immunoglobulin chain.

34. The isolated nucleic acid molecule of claim 30, wherein the variant B7-DC polypeptide is a variant of murine B7-DC.

35. The isolated nucleic acid molecule of claim 30, wherein the variant B7-DC polypeptide is a variant of human B7-DC.

36. The isolated nucleic acid molecule of claim 30, wherein the variant B7-DC polypeptide or fragment thereof has decreased binding affinity for PD-1 compared to wild-type B7-DC polypeptide.

37. The isolated nucleic acid molecule of claim 36, wherein the variant B7-DC polypeptide or fragment thereof enhances an immune response in a mammal.

38. The isolated nucleic acid molecule of claim 30, wherein the variant B7-DC polypeptide or fragment thereof has increased binding affinity for PD-1 compared to wild-type B7-DC polypeptide.

39. The isolated nucleic acid molecule of claim 38, wherein the variant B7-DC polypeptide or fragment thereof enhances an immune response in a mammal.

40. An isolated nucleic acid molecule encoding a fusion polypeptide comprising the IgV domain of B7-DC fused to a second polypeptide, wherein the IgV domain of B7-DC is from human (SEQ ID NO:

5) or murine B7-DC (SEQ ID NO;

6) and comprises an amino acid substitution relative to wildtype human (SEQ ID NO;

5) or murine (SEQ ID NO;

6) B7-DC IgV domain, wherein the substitution alters binding affinity for PD-1 compared to the wild-type B7-DC polypeptide.

41. The isolated nucleic acid of claim 40, wherein the second polypeptide comprises an amino acid sequence corresponding to the hinge, CH2 and CH3 regions of a human immunoglobulin chain.

42. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 111 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

43. The isolated nucleic acid molecule of claim 42, wherein the substitution is with a serine residue.

44. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 113 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

45. The isolated nucleic acid molecule of claim 44, wherein the substitution is with a serine residue.

46. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 56 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

47. The isolated nucleic acid molecule of claim 46, wherein the substitution is with a serine residue.

48. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 67 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

49. The isolated nucleic acid molecule of claim 48, wherein the substitution is with a tyrosine residue.

50. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 71 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

51. The isolated nucleic acid molecule of claim 50, wherein the substitution is with a serine residue.

52. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 101 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

53. The isolated nucleic acid molecule of claim 52, wherein the substitution is with a serine residue.

54. The isolated nucleic acid molecule of claim 41, wherein the IgV domain of B7-DC comprises an amino acid substitution at position 105 numbered from the initiation methionine of wild-type murine B7-DC (SEQ ID NO:

6) or human B7-DC (SEQ ID NO;

5).

55. The isolated nucleic acid molecule of claim 54, wherein the substitution is with an alanine.

56. A vector comprising the nucleic acid molecule of claim 41.

57. The vector of claim 56, wherein the nucleic acid molecule is operably linked to an expression control sequence.

58. An isolated host cell comprising the vector of claim 57.

59. A method for enhancing an immune response in a mammal comprising administering to the mammal the vector of claim 57.

60. A method for enhancing an immune response in a mammal comprising administering to the mammal a polypeptide encoded by a polynucleotide of claim 40.

61. A method for enhancing an immune response in a mammal comprising administering to the mammal a polypeptide produced by the host cell of claim 58.