Methods of producing enriched populations of tumor reactive T cells from peripheral blood

• US 9,844,569 B2
• Filed: 04/30/2013
• Issued: 12/19/2017
• Est. Priority Date: 03/01/2013
• Status: Active Grant

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: The United States of America, as represented by the Secretary, Department of Health and Human Services
• Inventors: Gros, Alena, Rosenberg, Steven A.
• Primary Examiner(s): Xiao, Yan
• Application Number:
  US14/771,593
• Publication Number:
  US 20160008400A1
• Time in Patent Office: 1,694 Days
• Field of Search: 424 937, 435378
• 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 39/4644 Cancer antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/56972 White blood cells

Abstract

Methods of obtaining a cell population enriched for tumor-reactive T cells, the method comprising: (a) obtaining a bulk population of peripheral blood mononuclear cells (PBMCs) from a sample of peripheral blood; (b) specifically selecting CD8⁺T cells that also express PD-1 and/or TIM-3 from the bulk population; and (c) separating the cells selected in (b) from unselected cells to obtain a cell population enriched for tumor-reactive T cells are disclosed. Related methods of administering a cell population enriched for tumor-reactive T cells to a mammal, methods of obtaining a pharmaceutical composition comprising a cell population enriched for tumor-reactive T cells, and isolated or purified cell populations are also disclosed.

Claims

1. A method of administering a cell population enriched for tumor-reactive T cells to a mammal, the method comprising:

(a) obtaining a bulk population of peripheral blood mononuclear cells (PBMCs) from a sample of peripheral blood;

(b) specifically selecting CD8⁺ T cells that also express PD-1 and/or TIM-3 from the bulk population;

(c) separating the cells selected in (b) from unselected cells to obtain a cell population enriched for tumor-reactive T cells; and

(d) administering the cell population enriched for tumor-reactive T cells to the mammal.

2. The method of claim 1, wherein (b) further comprises specifically selecting CD8⁺ T cells that express PD-1 from the bulk population.

4. The method of claim 1, wherein the cell population enriched for tumor-reactive T cells is obtained without screening for autologous tumor recognition.

5. The method of claim 1, wherein the bulk population of T cells is not nonspecifically stimulated prior to (b).

6. The method of claim 1, further comprising expanding the numbers of T cells in the enriched cell population obtained in (c).

7. The method of claim 1, further comprising culturing the enriched cell population obtained in (c) in the presence of any one or more of TWS119, interleukin (IL-21), IL-12, IL-15, IL-7, transforming growth factor (TGF) beta, and AKT inhibitor (AKTi).

8. The method of claim 1, further comprising stimulating the enriched cell population obtained in (c) with a tumor antigen and/or with autologous tumor T cell.

9. The method of claim 1, further comprising transducing or transfecting the cells of the enriched population obtained in (c) with a nucleotide sequence encoding any one or more of IL-12, IL-7, IL-15, IL-2, IL-21, mir155, and anti-PD-1 siRNA.

10. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 1 in an amount effective to treat cancer in the mammal.

11. The method of claim 1, wherein (b) comprises specifically selecting CD8⁺ T cells that are TIM-3⁺/PD-1⁺ from the bulk population.

12. The method of claim 1, wherein (b) comprises specifically selecting CD8⁺ T cells that are TIM-3⁺PD-1−

from the bulk population.

20. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 2 in an amount effective to treat cancer in the mammal.

21. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 4 in an amount effective to treat cancer in the mammal.

22. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 5 in an amount effective to treat cancer in the mammal.

23. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 11 in an amount effective to treat cancer in the mammal.

24. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 12 in an amount effective to treat cancer in the mammal.

3. A method of administering a cell population enriched for tumor-reactive T cells to a mammal, the method comprising:

(a) obtaining a bulk population of peripheral blood mononuclear cells (PBMCs) from a sample of peripheral blood;

(b) specifically selecting CD8⁺ T that are (i) TIM-3⁺/PD-1⁺, (ii) TIM-3⁻

/PD-1⁺, or (iii) TIM-3⁺/PD-1⁻

from the bulk population;

(c) separating the cells selected in (b) from unselected cells to obtain a cell population enriched for tumor-reactive T cells; and

(d) administering the cell population enriched for tumor-reactive T cells to the mammal.

13. The method of claim 3, wherein the cell population enriched for tumor-reactive T cells is obtained without screening for autologous tumor recognition.

14. The method of claim 3, wherein the bulk population of T cells is not nonspecifically stimulated prior to (b).

15. The method of claim 3, further comprising expanding the numbers of T cells in the enriched cell population obtained in (c).

16. The method of claim 3, further comprising culturing the enriched cell population obtained in (c) in the presence of any one or more of TWS119, interleukin (IL-21), IL-12, IL-15, IL-7, transforming growth factor (TGF) beta, and AKT inhibitor (AKTi).

17. The method of claim 3, further comprising stimulating the enriched cell population obtained in (c) with a tumor antigen and/or with autologous tumor T cell.

18. The method of claim 3, further comprising transducing or transfecting the cells of the enriched population obtained in (c) with a nucleotide sequence encoding any one or more of IL-12, IL-7, IL-15, IL-2, IL-21, mir155, and anti-PD-1 siRNA.

19. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 3 in an amount effective to treat cancer in the mammal.

25. A method of treating cancer in a mammal, the method comprising administering a cell population to the mammal by the method of claim 13 in an amount effective to treat cancer in the mammal.