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

Patent metadata

Field	Value
Publication number	US-9844569-B2
Application number	US-201314771593-A
Country	US
Kind code	B2
Filing date	Apr 30, 2013
Priority date	Mar 1, 2013
Publication date	Dec 19, 2017
Grant date	Dec 19, 2017

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.

First claim

The invention claimed is:

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.

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.

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.

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.

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.

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.

Assignees

• Us Health

Inventors

• Gros Alena
• Rosenberg Steven A

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• G01N33/56972

  White blood cells · CPC title

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title

• G01N33/5094

  for blood cell populations (red blood cells G01N33/80) · CPC title

• A61K35/17 Primary

  Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) · CPC title