Allogeneic cell therapy: mirror effect

• US 20040228848A1
• Filed: 05/04/2004
• Published: 11/18/2004
• Est. Priority Date: 05/13/2003
• Status: Active Grant

• Current Assignee: Mirror Biologics, Inc.
• Original Assignee: Immunovative Therapies Ltd
• Inventors: Har-Noy, Michael
• Granted Patent: US 7,435,592 B2
• Time in Patent Office: Days
• US Class Current: 424/93.71
• CPC Class Codes: A61K 2039/5154 Antigen presenting cells [APCs], e.g. dendritic cells or macrophages
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 2239/26 Universal/off- the- shelf cellular immunotherapy; Allogenic cells or means to avoid rejection
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 39/0005 Vertebrate antigens from snakes A61K39/38
• 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 39/4637 Other peptides or polypeptides
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C12N 2501/51 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 2531/00 Microcarriers
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 5/0636 : T lymphocytes

Abstract

A method of manipulating allogeneic cells for use in allogeneic cell therapy protocols is described. The method provides a composition of highly activated allogeneic T-cells which are infused into immunocompetent cancer patients to elicit a novel anti-tumor immune mechanism called the “Mirror Effect”. In contrast to current allogeneic cell therapy protocols where T-cells in the graft mediate the beneficial graft vs. tumor (GVT) and detrimental graft vs. host (GVH) effects, the allogeneic cells of the present invention stimulate host T-cells to mediate the “mirror” of these effects. The mirror of the GVT effect is the host vs. tumor (HVT) effect. The “mirror” of the GVH effect is the host vs. graft (HVG) effect. The effectiveness and widespread application of the anti-tumor GVT effect is limited by the severe toxicity of the GVH effect. In the present invention, the anti-tumor HVT effect occurs in conjunction with a non-toxic HVG rejection effect. The highly activated allogeneic cells of the invention can be used to stimulate host immunity in a complete HLA mis-matched setting in patients that have not had a prior bone marrow transplant or received chemotherapy and/or radiation conditioning regimens.

Claims

1. A method for stimulating a host immune system, the method comprising:

(a) Collecting mononuclear cells from a donor;

(b) Activating T-cells within said mononuclear cells ex-vivo;

(c) Administering said activated T-cells to a host having a host immune system, whereby the activated T-cells are rejected by the host immune system while stimulating the host immune system to mediate an effective immune response.

2. The method of claim 1 wherein said host has cancer.

3. The method of claim 2 wherein said host has a hematological malignancy.

4. The method of claim 2 wherein said cancer is a solid tumor.

5. The method of claim 4 wherein said solid tumor has metastasized.

6. The method of claim 1 wherein said donor is selected without regard to histocompatibility to said host.

7. The method of claim 1 wherein said donor is selected for maximum histocompatibility mis-match with said host.

8. The method of claim 1 wherein said host has not had a prior bone marrow transplant procedure.

9. The method of claim 1 wherein said host has not received any immunosuppressive chemotherapy and/or radiation conditioning regimens designed to allow engraftment of allogeneic donor cell infusions.

10. The method of claim 1 wherein said T-cells are CD4+ T-cells.

11. The method of claim 10 wherein a majority of said CD4+ T-cells differentiate after ex-vivo activation from CD45RA+, CD62L^hi naive cells into CD45RO+, CD62L^lo memory cells.

12. The method of claim 11 wherein said CD4+ T-cells produce at least one of the following cytokines:

IL-2, IFN-gamma, TNF-alpha.

13. The method of claim 10 wherein said CD4+ T-cells after ex-vivo activation express CD40L and/or TRAIL on the cell surface.

14. The method of claim 1 wherein said T-cells are activated by cross-linking of anti-CD3 and anti-CD28 mAbs applied to the cell surface of the T-cells.

15. The method of claim 1 wherein greater than 90% of the T-cells are activated at the time of administration to the host and just prior to contacting the host immune system.

16. The method of claim 1 wherein greater than 95% of the T-cells are activated at the time of administration to the host and just prior to contacting the host immune system.

17. The method of claim 14 wherein said anti-CD3 and anti-CD28 mAbs applied to the surface of said T-cells are cross-linked by association with biodegradable microspheres coated with an agent reactive against said mAbs.

18. The method of claim 1 wherein said T-cells are continuously exposed to an activating stimulus for at least 6 days.

19. The method of claim 1 wherein said T-cells are activated while being maintained at cell densities of at least 10⁷ cells/ml to maximize cell to cell contact.

20. The method of claim 18 wherein said T-cells are activated while being maintained at cell densities of at least 10⁷ cells/ml to maximize cell to cell contact.

21. The method of claim 1 wherein said T-cells are administered with anti-CD3 and anti-CD28 mAbs applied to the surface of the T-cells and wherein said mAbs are cross-linked by association with and inclusion of biodegradable microspheres coated with an agent reactive against said mAbs.

22. The method of claim 1 wherein said activated T-cell administration stimulates production of Type 1 cytokines.

23. The method of claim 22 wherein said Type 1 cytokines include at least one of the following:

IL-1, IL-2, IL-12, IL-15, IFN-gamma, IFN-alpha, IFN-beta, TNF-alpha, and TNF-beta.

24. The method of claim 22 wherein said Type 1 cytokines stimulates host innate immune function.

25. The method of claim 1 wherein said activated T-cell administration activates host dendritic and/or macrophage cells.

26. The method of claim 1 wherein said activated T-cell administration and subsequent rejection of said activated T-cells stimulates an immune response against a host resident disease.

27. The method of claim 1 wherein said ex-vivo activated T-cells are cryopreserved prior to administration to the host.

28. A composition of T-cells labeled with anti-CD3 and anti-CD28 mAbs cross-linked with biodegradable microspheres coated with an agent reactive against said mAbs.

29. The composition of claim 28 wherein said labeled T-cells and associated biodegradable microspheres are suspended in a media suitable for intravenous infusion.

30. The composition of claim 29 wherein said labeled T-cells and associated biodegradable microspheres are suspended at a cell density of 10⁷ cells/ml or greater.

31. The composition of claim 30 wherein said labeled T-cells and associated biodegradable microspheres are suspended at a cell density of 10⁷ cells/ml or greater in a flexible container.

32. The composition of claim 30 wherein said labeled T-cells and associated biodegradable microspheres are suspended at a cell density of 10⁷ cells/ml or greater in a syringe.

33. The composition of claim 28 wherein the composition is cryopreserved.

34. An allogeneic cell material that elicits a host vs. tumor and host vs. graft response when contacted with a tumor-bearing host immune system without eliciting a graft vs. host response.

35. The allogeneic cell material of claim 34 wherein the said allogeneic cell material contains ex-vivo activated T-cells.

36. The allogeneic cell material of claim 35 wherein said activated T-cells are CD4+ T-cells.

37. An allogeneic cell material that causes apoptosis of tumors when administered to a tumor-bearing host.

38. The allogeneic cell material of claim 37 that contains activated T-cells.

39. The allogeneic cell material of claim 38 wherein the activated T-cells are CD4+ cells.

40. The allogeneic cell material of claim 39 wherein the CD4+ cells express FasL and/or TRAIL.

41. The allogeneic cell material of claim 37 wherein said activated T-cells differentiate into memory cells expressing CD45RO and CD62L^lo.

42. The allogeneic cell material of claim 37 wherein said activated CD4+ cells express one or more of the following cytokines:

IL-2, IL-15, IFN-gamma, TNF-alpha.

43. The allogeneic cell material of claim 37 wherein said activated T-cells express FasL.

44. The allogeneic cell material of claim 37 wherein said activated T-cells express TRAIL.

45. A composition comprising a treatment effective amount of a population of allogeneic cells, of which at least a portion are T-cells, and whereby said T-cells are labeled with an activating effective amount of one or more monoclonal antibodies, or portions thereof, and a cross-linking effective amount of an agent reactive against said monoclonal antibodies.

46. The composition of claim 45 wherein the T-cells are labeled with anti-CD3 and anti-CD28 mAbs.

47. The composition of claim 45 wherein the agent reactive against said mAbs is coated on biodegradable microspheres.

48. The composition of claim 47 wherein the T-cells and associated biodegradable microspheres are suspended in a media suitable for intravenous infusion.

49. The composition of claim 48 wherein the T-cells and associated biodegradable microspheres are suspended at a cell density of 10⁷ cells/ml or greater.

50. The composition of claim 49 wherein the T-cells and associated biodegradable microspheres are suspended at a cell density of 10⁷ cells/ml or greater in a flexible container.

51. The composition of claim 49 wherein the T-cells and associated biodegradable microspheres are suspended at a cell density of 10⁷ cells/ml or greater in a syringe.

52. The composition of claim 45 wherein the composition is cryopreserved.