Expansion of T cells in vitro and expanded T cell populations

• US 20030153073A1
• Filed: 11/07/2002
• Published: 08/14/2003
• Est. Priority Date: 11/07/2001
• Status: Abandoned Application

• Current Assignee: Paul Rodgers, Toshifumi Mikayama
• Original Assignee: Paul Rodgers, Toshifumi Mikayama
• Inventors: Rogers, Paul, Mikayama, Toshifumi
• Application Number:
  US10/291,809
• Publication Number:
  US 20030153073A1
• Time in Patent Office: Days
• US Class Current: 435/372
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• 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 39/4613 Natural-killer cells [NK or NK-T]
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 33/06 : Antimalarials
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C12N 2501/23 : Interleukins [IL]
• CPC Class Codes: C12N 2502/11 : blood or immune system cells
• CPC Class Codes: C12N 5/0636 : T lymphocytes

Abstract

Methods for expanding T cells (e.g., Vα24⁺Vβ11⁺ human NKT cells ) in vitro by repeated culturing in the presence of an antigen (e.g., glycosphingolipid), antigen presenting cells, and a cell survival factor are provided. A population of NKT cells (Vα24⁺ζβ11⁺) can be expanded more than 1 million-fold in 5 weeks and more than 1 billion-fold in 10 weeks.

Claims

1. A method for stimulating the proliferation of human T cells in vitro comprising repeatedly culturing donor T cells in the presence of antigen presenting cells, an antigen, a cell survival factor and serum for at least 7 days under conditions stimulating proliferation of the T cells.

2. The method of claim 1, wherein at least a portion of the proliferated T cells express Vα

24 T cell receptor (TCR).

3. The method of claim 1, wherein at least a portion of the proliferated T cells express CD3 or CD161.

4. The method of claim 1, wherein at least a portion of the proliferated T cells are capable of killing a cell

5. The method of claim 1, wherein the antigen presenting cells are from the same human as the human donor T cells or are from a different human.

6. The method of claim 1, wherein the antigen presenting cells are human or are non-human.

7. The method of claim 1, wherein the antigen presenting cells are engineered to express human or non-human CD1a, CD1b, CD1c or CD1d, or a molecule having the glycolipid binding activity of human or non-human CD1a, CD1b, CD1c or CD1d.

8. The method of claim 1, wherein the antigen presenting cells are present in or obtained from human peripheral blood mononuclear cells (PBMC).

9. The method of claim 1, wherein the cells are passaged at least five times.

10. The method of claim 1, wherein the serum is human.

11. The method of claim 1, wherein the serum is non-human.

12. The method of claim 1, wherein the serum is replaced with a serum-free medium.

13. The method of claim 1, wherein the cell survival factor comprises a molecule that binds to a molecule on the surface of a T cell.

14. The method of claim 1, wherein the cell survival factor comprises IL-2.

15. The method of claim 1, wherein the cell survival factor comprises IL-2, IL-7 or Il-15.

16. The method of claim 1, wherein the cells proliferate to about 10⁸ cells or greater over 10 weeks.

17. The method of claim 1, wherein the antigen comprises a glycosphingolipid, a bacterial antigen or a viral antigen.

18. The method of claim 17, wherein the glycosphingolipid comprises KRN 7000 or a KRN 7000 analogue.

19. The method of claim 18, wherein the KRN7000 analog comprises β

-glucosylceramide (β

-GluCer).

20. The method of claim 17, wherein the bacterial antigen comprises tetanus toxoid, diphtheria toxin, BCG, pertussis antigen, Hemophilus influenzae type B antigen or a pneumoccoccol antigen.

21. The method of claim 17, wherein the viral antigen comprises a measles virus antigen, rubella viral antigen, varicella viral antigen, or hepatitis viral antigen.

22. The method of claim 1, wherein the donor T cells are antigen sensitized.

23. The method of claim 1, wherein at least a portion of the proliferated T cells produce one or more cytokines.

24. The method of claim 1, wherein at least a portion of the proliferated T cells exhibit anti-tumor cell activity.

25. The method of claim 1, wherein at least a portion of the proliferated T cells activate NK cells to exhibit anti-tumor cell activity.

26. The method of claim 1, wherein the antigen presenting cells are non-viable.

27. The method of claim 1, wherein the antigen presenting cells are irradiated.

52. A proliferated T cell culture produced by the method of claims 1 or 28.

28. A method for stimulating the proliferation of human T cells in vitro comprising repeatedly culturing donor T cells in the presence of antigen presenting cells, an antigen, IL-2, without IL-7 or IL-15, and serum under conditions stimulating proliferation of the T cells.

29. The method of claim 28, wherein at least a portion of the proliferated T cells express Vα

24 T cell receptor (TCR).

30. The method of claim 28, wherein at least a portion of the proliferated T cells express CD3 or CD161.

31. The method of claim 28, wherein at least a portion of the proliferated T cells are capable of killing a cell

32. The method of claim 28, wherein the antigen presenting cells are from the same human as the human donor T cells or are from a different human.

33. The method of claim 28, wherein the antigen presenting cells are human or are non-human.

34. The method of claim 28, wherein the antigen presenting cells are engineered to express human or non-human CD1a, CD1b, CD1c or CD1d, or a molecule having the glycolipid binding activity of human or non-human CD1a, CD1b, CD1c or CD1d.

35. The method of claim 28, wherein the antigen presenting cells are present in or obtained from human peripheral blood mononuclear cells (PBMC).

36. The method of claim 28, wherein the cells are passaged at least five times.

37. The method of claim 28, wherein the serum is human.

38. The method of claim 28, wherein the serum is non-human.

39. The method of claim 28, wherein the serum is replaced with a serum-free medium.

40. The method of claim 28, wherein the cells proliferate to about 10⁸ cells or greater over 10 weeks.

41. The method of claim 28, wherein the antigen comprises a glycosphingolipid, a bacterial antigen or a viral antigen.

42. The method of claim 41, wherein the glycosphingolipid comprises KRN 7000 or a KRN 7000 analogue.

43. The method of claim 42, wherein the KRN7000 analog comprises β

-glucosylceramide (β

-GluCer).

44. The method of claim 42, wherein the bacterial antigen comprises tetanus toxoid, diphtheria toxin, BCG, pertussis antigen, Hemophilus influenzae type B antigen or a pneumoccoccol antigen.

45. The method of claim 42, wherein the viral antigen comprises a measles virus antigen, rubella viral antigen, varicella viral antigen, or hepatitis viral antigen.

46. The method of claim 28, wherein the donor T cells are antigen sensitized.

47. The method of claim 28, wherein at least a portion of the proliferated T cells produce one or more cytokines.

48. The method of claim 28, wherein at least a portion of the proliferated T cells exhibit anti-tumor cell activity.

49. The method of claim 28, wherein at least a portion of the proliferated T cells activate NK cells to exhibit anti-tumor cell activity.

50. The method of claim 28, wherein the antigen presenting cells are non-viable.

51. The method of claim 28, wherein the antigen presenting cells are irradiated.

53. A method for providing cell therapy to a subject comprising administering to the subject a T cell culture of claim 28 in an amount sufficient to provide therapy to the subject.

54. The method of claim 53, wherein the donor T cells are obtained from the subject to which the proliferated T cells are administered.

55. The method of claim 53, wherein the subject has or is at risk of having undesirable numbers of T cells.

56. The method of claim 53, wherein the subject has or is at risk of having undesirable numbers of T cells that express Vα

4 T cell receptor.

57. The method of claim 53, wherein the subject is a candidate for or has undergone organ or tissue transplant.

58. The method of claim 53, wherein the subject has or is at risk of having an immune deficiency, an autoimmune disorder, a cancer, or an infectious disease.

59. The method of claim 58, wherein the immune deficiency is associated with an organ or tissue transplant.

60. The method of claim 58, wherein the autoimmune disorder is selected from:

diabetes, multiple sclerosis, systemic sclerosis, colitis, hepatitis, lupus, rheumatoid arthritis or Sjö

gren'"'"'s syndrome.

61. The method of claim 58, wherein the cancer comprises a solid tumor, metastatic tumor, leukemia, lymphoma or myeloma.

62. The method of claim 61, wherein the leukemia comprises T cell, B cell or monocytic leukemia.

63. The method of claim 61, wherein the lymphoma comprises Hodgkin'"'"'s lymphoma or non-Hodgkin'"'"'s lymphoma.

64. The method of claim 58, wherein the cancer comprises an adenocarcinoma, plasmacytoma, sarcoma, carcinoma or neuroblastoma.

65. The method of claim 58, wherein the infectious disease is caused by a virus, bacterium, fungus, or a parasite.

66. The method of claim 65, wherein the virus comprises human immunodeficiency virus (HIV), hepatitis C virus (HCV), cytomegalovirus or hepatitis B virus (HBV).

67. The method of claim 65, wherein the bacterium causes tuberculosis, lyme disease or leprosy.

68. The method of claim 65, wherein the parasite causes malaria or Chagas'"'"' disease.

69. A method for producing a population of proliferated human T cells, comprising repeatedly culturing donor T cells in the presence of antigen presenting cells, an antigen, a cell survival factor and serum for at least 7 days, thereby producing a population of proliferated human T cells.

70. The method of claim 69, wherein the cell survival factor comprises IL-2.

71. The method of claim 69, wherein the cell survival factor comprises IL-2 without IL-7 or IL-15.

72. A method for producing a population of proliferated human T cells, comprising repeatedly culturing donor T cells in the presence of antigen presenting cells, an antigen, IL-2, without IL-7 or IL-15, and serum, thereby producing a population of proliferated human T cells.

73. The method of claim 72, wherein the donor T cells are cultured for at least 7 days.