Methods for treating HIV infected subjects

• US 7,232,566 B2
• Filed: 01/04/2005
• Issued: 06/19/2007
• Est. Priority Date: 11/23/1988
• Status: Expired due to Fees

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.), The United States As Represented By The Secretary of The Navy, Board of Regents of the University of Michigan (University of Michigan)
• Inventors: Nabel, Gary J., June, Carl H., Thompson, Craig B., Gray, Gary S., Rennert, Paul D.
• Primary Examiner(s): Gambel; Phillip
• Application Number:
  US11/029,188
• Publication Number:
  US 20060013832A1
• Time in Patent Office: 896 Days
• Field of Search: None
• US Class Current: 424/93.71
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/02 Bacterial antigens
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/464838 Viral antigens
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 14/70532 B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2806 against CD2
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2812 against CD4
• CPC Class Codes: C07K 16/2815 against CD8
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 16/2833 against MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/289 against CD45
• CPC Class Codes: C07K 16/2896 : against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/54 : F(ab')2
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/74 : Inducing cell proliferation
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• 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
• CPC Class Codes: C12N 2501/51 : B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand)
• CPC Class Codes: C12N 2501/515 : CD3, T-cell receptor complex
• CPC Class Codes: C12N 2501/599 : with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2710/00088 : for redistribution
• CPC Class Codes: C12N 2740/16011 : Human Immunodeficiency Virus, HIV
• CPC Class Codes: C12N 5/0636 : T lymphocytes

Abstract

Methods for inducing a population of T cells to proliferate by activating the population of T cells and stimulating an accessory molecule on the surface of the T cells with a ligand which binds the accessory molecule are described. T cell proliferation occurs in the absence of exogenous growth factors or accessory cells. T cell activation is accomplished by stimulating the T cell receptor (TCR)/CD3 complex or the CD2 surface protein. To induce proliferation of an activated population T cells, an accessory molecule on the surface of the T cells, such as CD28, is stimulated with a ligand which binds the accessory molecule. The T cell population expanded by the method of the invention can be genetically transduced and used for immunotherapy or can be used in methods of diagnosis.

Claims

1. A method for expanding a population of CD4⁺T cells from an HIV-infected individual to sufficient numbers for use in therapy, comprising:

a) contacting the population of CD4⁺T cells from an HIV-infected individual ex vivo with a surface comprising;

(1) a first agent which provides a primary activation signal, thereby activating the CD4⁺T cells, wherein the first agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the CD4⁺T cell when complexed with the TCR/CD3 complex; and

(2) a second agent which stimulates a CD28 antigen, thereby inducing proliferation of the activated CD4⁺T cells, wherein the second agent is selected from the group consisting of an anti-CD28 antibody or a CD28-binding fragment thereof, B7-1, a CD28-binding fragment of B7-1, B7-2, a CD28-binding fragment of B7-2, a B7-1Ig fusion protein that interacts with CD28, and a B7-2Ig fusion protein that interacts with CD28,

wherein the first agent and second agent are attached on the same surface, thereby expanding the CD4⁺T cells to sufficient numbers for use in therapy.

2. The method of claim 1, further comprising culturing the population of CD4⁺T cells in the presence of IL-2.

3. The method of claim 1, further comprising culturing the population of CD4⁺T cells in the presence of an anti-retroviral agent.

4. The method of claim 1, further comprising:

b) monitoring proliferation of the CD4⁺T cells; and

c) reactivating and restimulating the CD4⁺T cells when the rate of CD4⁺T cell proliferation has decreased to induce further proliferation of the CD4⁺T cells.

5. The method of claim 1, wherein the expanded population of CD4⁺T cells is administered to an HIV-infected patient.

6. The method of claim 1, wherein the expanded population of CD4⁺T cells contains at least about 10¹⁰ T cells.

7. The method of claim 1, wherein the expanded population of CD4⁺T cells contains at least about 10¹² T cells.

8. The method of claim 1, wherein the anti-CD3 and the anti-CD28 antibodies or fragments thereof are monoclonal antibodies or fragments thereof.

9. The method of claim 1, wherein said surface is selected from the group consisting of a bead, a cell surface, and a tissue culture dish.

10. The method of claim 1, wherein the first agent and second agent are directly attached on the same surface.

11. The method of claim 1, wherein the first agent and second agent are indirectly attached on the same surface.

12. The method of claim 1, wherein the first agent and second agent are covalently attached on the same surface.

13. The method of claim 1, wherein the T cells are induced to proliferate to about 100-fold the original T cell population.

14. The method of claim 1, wherein the T cells are induced to proliferate to about 100,000-fold the original T cell population.

15. The method of claim 1, wherein the first agent is an anti-CD3 antibody or a CD3-binding fragment thereof.

16. The method of claim 1, wherein the first agent is an anti-CD2 antibody or a CD2-binding fragment thereof.

17. The method of claim 1, wherein the first agent is an antigen in a form suitable to trigger a primary activation signal in the CD4⁺T cell when complexed with the TCR/CD3 complex.

18. The method of claim 1, wherein the second agent is an anti-CD28 antibody or a CD28-binding fragment thereof.

19. The method of claim 1, wherein the second agent is B7-1.

20. The method of claim 1, wherein the second agent is a CD28-binding fragment of B7-1.

21. The method of claim 1, wherein the second agent is B7-2.

22. The method of claim 1, wherein the second agent is a CD28-binding fragment of B7-2.

23. The method of claim 1, wherein the second agent is a B7-1Ig fusion protein that interacts with CD28.

24. The method of claim 1, wherein the second agent is a B7-2Ig fusion protein that interacts with CD28.

25. The method of claim 9, wherein the surface is a bead.

26. The method of claim 9, wherein the surface is a cell surface.

27. The method of claim 9, wherein the surface is a tissue culture dish.

28. The method of claim 25, wherein the bead is a magnetic immunobead.

29. The method of claim 15, wherein the anti-CD3 monoclonal antibody or the CD3-binding fragment thereof is OKT3 or a CD3-binding fragment thereof.

30. The method of claim 18, wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is EX5.3D10 or a CD28-binding fragment thereof obtainable from a hybridoma deposited as ATCC Deposit No. HB11373.

31. The method of claim 18, wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is 9.3 or a CD28-binding fragment thereof obtainable from a hybridoma deposited as ATCC Deposit No. HB10271.

32. The method of claim 23, wherein the B7-1Ig fusion protein comprises amino acids 1-208 of SEQ ID NO:

2.

33. The method of claim 23, wherein the Ig component of the B7-1Ig fusion protein is selected from the group consisting of an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG1, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG2, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG3 and an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG4.

34. The method of claim 24, wherein the B7-2Ig fusion protein comprises amino acids 24-245 of SEQ ID NO:

4.

35. The method of claim 24, wherein the Ig component of the B7-2Ig fusion protein is selected from the group consisting of an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG1, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG2, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG3 and an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG4.

36. A method for expanding a population of CD4⁺ T cells from an HIV-infected individual in number of from about 100 to about 100,000 fold over the original CD4⁺ T cell population or to about 10 log₁₀ to 12 log₁₀, comprising:

a) contacting a population of CD4⁺ T cells from the HIV-infected individual ex vivo with a surface comprising;

1) a first agent which provides a primary activation signal, thereby activating the CD4⁺ T cells, wherein the first agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and

2) a second agent which stimulates a CD28 antigen, thereby inducing proliferation of the activated CD4⁺ T cells, wherein the second agent is selected from the group consisting of an anti-CD28 antibody or a CD28-binding fragment thereof, B7-1 or a CD28-binding fragment thereof, B7-2 or a CD28-binding fragment thereof, a B7-1Ig fusion protein that interacts with CD28, and a B7-2Ig fusion protein that interacts with CD28,

wherein said first agent and said second agent are attached on the same surface;

b) adding exogenous IL-2 to the population of CD4⁺ T cells from the HIV-infected individual, such that the population of CD4⁺ T cells from the HIV-infected individual increases in number from about 100 to about 100,000 fold over the original CD4⁺ T cell population or to about 10 log₁₀ to 12 log₁₀.

37. The method of claim 36, wherein the second agent is an anti-CD28 monoclonal antibody or a CD28-binding fragment thereof.

38. The method of claim 37, wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is a human anti-CD28 antibody or a CD28-binding fragment thereof.

39. The method of claim 36, wherein IL-2 is added to the population of T cells in an amount sufficient to maintain proliferation of the T cells.

40. The method of claim 36, wherein IL-2 is added to the population of T cells from the beginning of the culture and supplemented every other day in an amount sufficient to maintain proliferation of the T cells.

41. The method of claim 36, wherein said surface is selected from the group consisting of a bead, a cell surface, and a tissue culture dish.

42. The method of claim 36, wherein the first agent and the second agent are covalently attached on the same surface.

43. The method of claim 36, wherein the first agent and the second agent are directly attached on the same surface.

44. The method of claim 36, wherein the first agent and the second agent are attached on the same surface via an avidin-biotin complex.

45. The method of claim 37, wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is EX5.3D10 or a CD28-binding fragment thereof obtainable from a hybridoma deposited as ATCC Deposit No. HB11373.

46. The method of claim 37, wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is 9.3 or a CD28-binding fragment thereof obtainable from a hybridoma deposited as ATCC Deposit No. HB10271.

47. The method of claim 36, wherein the first agent is an anti-CD3 antibody or a CD3-binding fragment thereof.

48. The method of claim 36, wherein the first agent is an anti-CD2 antibody or a CD2-binding fragment thereof.

49. The method of claim 36, wherein the first agent is an antigen in a form suitable to trigger a primary activation signal in the CD4⁺T cell when complexed with the TCR/CD3 complex.

50. The method of claim 36, wherein the second agent is a CD28-binding fragment of B7-1.

51. The method of claim 36, wherein the second agent is a CD28-binding fragment of B7-2.

52. The method of claim 36, wherein the second agent is a B7-1Ig fusion protein that interacts with CD28.

53. The method of claim 36, wherein the second agent is a B7-2Ig fusion protein that interacts with CD28.

54. The method of claim 41, wherein the surface is a bead.

55. The method of claim 41, wherein the surface is a cell surface.

56. The method of claim 41, wherein the surface is a tissue culture dish.

57. The method of claim 47, wherein the anti-CD3 antibody or the CD3-binding fragment thereof is a monoclonal antibody or a CD3-binding fragment thereof.

58. The method of claim 57, wherein the anti-CD3 monoclonal antibody or the CD3-binding fragment thereof is OKT3 or a CD3-binding fragment thereof.

59. The method of claim 54, wherein the bead is selected from the group consisting of a magnetic immunobead, and a polystyrene bead.

60. The method of claim 52, wherein the B7-1Ig fusion protein comprises amino acids 1-208 of SEQ ID NO:

2.

61. The method of claim 52, wherein the Ig component of the B7-1Ig fusion protein is selected from the group consisting of an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG1, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG2, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG3 and an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG4.

62. The method of claim 53, wherein the B7-2Ig fusion protein comprises about amino acids 24-245 of SEQ ID NO:

4.

63. The method of claim 53, wherein the Ig component of the B7-2Ig fusion protein is selected from the group consisting of an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG1, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG2, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG3 and an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG4.

64. A method for preparing a renewable source of CD4⁺T cells, comprising expanding a population of CD4⁺T cells according to the method of claim 36 and storing a portion of the cells under conditions which will not significantly affect their viability upon thawing.