Nucleic acids encoding costimulatory molecule B7-4

• US 6,936,704 B1
• Filed: 08/23/2000
• Issued: 08/30/2005
• Est. Priority Date: 08/23/1999
• Status: Expired due to Term

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Malenkovich, Nelly, Chernova, Tatyana, Freeman, Gordon, Boussiotis, Vassiliki
• Primary Examiner(s): Gambel, Phillip
• Assistant Examiner(s): Ouspenski, Ilia
• Application Number:
  US09/644,934
• Time in Patent Office: 1,833 Days
• Field of Search: 536/23.5, 536/23.1, 536/23.4, 435/455, 435/252.3, 435/320.1, 435/471, 435/326, 435/69.1, 435/255
• US Class Current: 536/23.5
• CPC Class Codes: A61K 2039/55516 Proteins; Peptides
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 14/70503 Immunoglobulin superfamily
• CPC Class Codes: C07K 14/70532 B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2827 against B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The invention provides isolated nucleic acids molecules, designated B7-4 nucleic acid molecules, which encode novel B7-4 polypeptides. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing B7-4 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a B7-4 gene has been introduced or disrupted. The invention still further provides isolated B7-4 proteins, fusion proteins, antigenic peptides and anti-B7-4 antibodies. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided.

Claims

1. An isolated nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:

1.

55. The isolated nucleic acid molecule of claim 1 which also encodes a heterologous polypeptide.

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

57. The vector of claim 56, which is an expression vector.

58. A host cell transfected with the expression vector of claim 57.

2. An isolated nucleic acid molecule encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

2.

60. The isolated nucleic acid molecule of claim 2 which also encodes a heterologous polypeptide.

61. A vector comprising the nucleic acid molecule of claim 2.

62. The vector of claim 61, which is an expression vector.

63. A host cell transfected with the expression vector of claim 62.

64. A method of producing a polypeptide encoded by the nucleic acid of claim 2, comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 2 in an appropriate culture medium, to thereby produce the polypeptide.

3. An isolated nucleic acid molecule comprising nucleotides 107-766 of SEQ ID NO:

3.

33. The isolated nucleic acid molecule of claim 3 which also encodes a heterologous polypeptide.

34. A vector comprising the nucleic acid molecule of claim 3.

35. The vector of claim 34, which is an expression vector.

36. A host cell transfected with the expression vector of claim 35.

37. A method of producing a polypeptide encoded by the nucleic acid of claim 3, comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 3 in an appropriate culture medium, to thereby produce the polypeptide.

4. An isolated nucleic acid molecule comprising a nucleotide sequence encoding amino acids 19-238 of SEQ ID NO:

4.

38. The isolated nucleic acid molecule of claim 4 which also encodes a heterologous polypeptide.

39. A vector comprising the nucleic acid molecule of claim 4.

40. The vector of claim 39, which is an expression vector.

41. A host cell transfected with the expression vector of claim 40.

42. A method of producing a polypeptide encoded by the nucleic acid of claim 4, comprising culturing a host cell transfected with an expression vector comprising a nucleic acid molecule of claim 4 in an appropriate culture medium, to thereby produce the polypeptide.

5. An isolated nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:

3.

6. The isolated nucleic acid molecule of claim 5 which also encodes a heterologous polypeptide.

7. A vector comprising the nucleic acid molecule of claim 5.

8. The vector of claim 7, which is an expression vector.

9. A host cell transfected with the expression vector of claim 8.

10. A method of producing a polypeptide encoded by the nucleic acid comprising the nucleotide sequence set forth in SEQ ID NO:

3, comprising culturing the host cell of claim 9 in an appropriate culture medium, to thereby produce the polypeptide.

11. An isolated nucleic acid molecule encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

4.

12. The isolated nucleic acid molecule of claim 11 which also encodes a heterologous polypeptide.

13. A vector comprising the nucleic acid molecule of claim 11.

14. The vector of claim 13, which is an expression vector.

15. A host cell transfected with the expression vector of claim 14.

16. A method of producing a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

4, comprising culturing the host cell of claim 15 in an appropriate culture medium, to thereby produce the polypeptide.

17. An isolated nucleic acid molecule comprising a nucleotide sequence which hybridizes to the complement of the nucleic acid molecule of SEQ ID NO:

3 over its full length under conditions of incubation at 45°

C. in 6×

sodium chloride/sodium citrate (SSC), followed by washing in 0.2×

SSC, 0.1% SDS, at 50-60°

C., wherein the nucleic acid molecule encodes a polypeptide that costimulates T cell proliferation in vitro when the polypeptide is present on a first surface and an antigen or a polyclonal activator that transmits an activating signal via the T-cell receptor is present on a second, different surface.

18. The isolated nucleic acid molecule of claim 17 which also encodes a heterologous polypeptide.

19. A vector comprising the nucleic acid molecule of claim 17.

20. The vector of claim 19, which is an expression vector.

21. A host cell transfected with the expression vector of claim 20.

22. A method of producing a polypeptide encoded by the nucleic acid of claim 17, comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 17 in an appropriate culture medium, to thereby produce the polypeptide.

23. An isolated nucleic acid molecule comprising a nucleotide sequence which hybridizes to the complement of the nucleic acid molecule of SEQ ID NO:

1 over its full length under conditions of incubation at 45°

C. in 6×

sodium chloride/sodium citrate (SSC), followed by washing in 0.2×

SSC, 0.1% SDS, at 50-60°

C., wherein the nucleic acid molecule encodes a polypeptide that costimulates T cell proliferation in vitro when the polypeptide is present on a first surface and an antigen or a polyclonal activator that transmits an activating signal via the T-cell receptor is present on a second, different surface.

24. The isolated nucleic acid molecule of claim 23 which also encodes a heterologous polypeptide.

25. A vector comprising the nucleic acid molecule of claim 23.

26. The vector of claim 25, which is an expression vector.

27. A host cell transfected with the expression vector of claim 26.

28. A method of producing a polypeptide encoded by the nucleic acid of claim 23, comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 23 in an appropriate culture medium, to thereby produce the polypeptide.

29. An isolated nucleic acid molecule comprising a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO:

3, and that encodes a polypeptide that costimulates T cell proliferation in vitro when the polypeptide is present on a first surface and an antigen or a polyclonal activator that transmits an activating signal via the T-cell receptor is present on a second, different surface.

30. An isolated nucleic acid molecule comprising a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

4, wherein the polypeptide costimulates T cell proliferation in vitro when the polypeptide is present on a first surface and an antigen or a polyclonal activator that transmits an activating signal via the T-cell receptor is present on a second, different surface.

31. An isolated nucleic acid molecule which is a fragment of SEQ ID NO:

1 or SEQ ID NO;

3, or the complement thereof, wherein the fragment consists of at least 15 contiguous nucleotides of a nucleotide sequence selected from the group consisting of;

(a) nucleotides 1-319 of SEQ ID NO;

1;

(b) nucleotides 53-922 of SEQ ID NO;

3;

(c) nucleotides 1-314 of SEQ ID NO;

3; and

(d) nucleotides 955-1285 of SEQ ID NO;

3.

32. An isolated nucleic acid molecule comprising a nucleotide sequence encoding the amino acids 19-245 of SEQ ID NO:

2.

43. An isolated nucleic acid molecule comprising nucleotides 107-733 of SEQ ID NO:

3.

44. The isolated nucleic acid molecule of claim 43 which also encodes a heterologous polypeptide.

45. A vector comprising the nucleic acid molecule of claim 43.

46. The vector of claim 45, which is an expression vector.

47. A host cell transfected with the expression vector of claim 46.

48. A method of producing a polypeptide encoded by the nucleic acid of claim 43, comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 43 in an appropriate culture medium, to thereby produce the polypeptide.

59. A method of producing a polypeptide encoded by the nucleic acid of claim 43 comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 43 in an appropriate culture medium, to thereby produce the polypeptide.

49. An isolated nucleic acid molecule comprising a nucleotide sequence encoding amino acids 19-227 of SEQ ID NO:

4.

50. The isolated nucleic acid molecule of claim 49 which also encodes a heterologous polypeptide.

51. A vector comprising the nucleic acid molecule of claim 49.

52. The vector of claim 51, which is an expression vector.

53. A host cell transfected with the expression vector of claim 52.

54. A method of producing a polypeptide encoded by the nucleic acid of claim 49, comprising culturing a host cell transfected with an expression vector comprising the nucleic acid molecule of claim 49 in an appropriate culture medium, to thereby produce the polypeptide.