METHODS OF PRODUCING T CELL POPULATIONS ENRICHED FOR STABLE REGULATORY T-CELLS

• US 20130157363A1
• Filed: 12/17/2012
• Published: 06/20/2013
• Est. Priority Date: 12/16/2011
• Status: Active Grant

• Current Assignee: The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: The United States of America As Represented By The Secretary of Agriculture
• Inventors: Kim, Yong Chan, Shevach, Ethan
• Granted Patent: US 9,481,866 B2
• Time in Patent Office: Days
• US Class Current: 435/375
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• 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: C12N 2501/056 Immunostimulating oligonucleotides, e.g. CpG
• CPC Class Codes: C12N 2501/515 CD3, T-cell receptor complex
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: C12N 5/0637 Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg

Abstract

The present invention provides methods for producing cell populations enriched for stable, regulatory T cells (Tregs). In particular, the invention relates to methods for culturing T cells such that the final culture is enriched for stable, regulatory T cells. It also relates to methods for stabilizing regulatory T cells. Also provided are compositions enriched for stable, regulatory T cells, which are useful for treating individuals in need of such treatment. The methods and compositions disclosed herein can also be used to treat an individual suffering from an immune-mediated disease.

Claims

1. A method for producing a population of cells having stable, regulatory T cells, the method comprising culturing isolated cells comprising an initial population of regulatory T-cells, in the presence of an oligonucleotide to expand the initial, regulatory T-cells.

2. The method of claim 1, wherein the isolated oligonucleotide is between 11 and 49 nucleotides in length.

3. The method of claim 1, wherein the isolated oligonucleotide is between 15 and 40 nucleotides in length.

4. The method of claim 1, wherein the isolated oligonucleotide is between 20 and 30 nucleotides in length.

5. The method of claim 1, wherein the isolated oligonucleotide is selected from the group consisting of an isolated oligonucleotide 21 nucleotides in length, an isolated oligonucleotide 22 nucleotides in length, an isolated oligonucleotide 23 nucleotides in length, an isolated oligonucleotide 24 nucleotides in length, and an isolated oligonucleotide 25 nucleotides in length.

6. The method of claim 1, wherein the isolated oligonucleotide is approximately 25 nucleotides length.

7. The method of claim 1, wherein the isolated cells are peripheral blood mononuclear cells.

8. The method of claim 1, wherein the isolated cells are lymphocytes.

9. The method of claim 1, wherein at least some of the initial regulatory T cells are CD4+ and Foxp3+.

10. The method of claim 9, wherein at least some of the initial regulatory T cells are also Helios+.

11. The method of claim 1, wherein at least some of the initial regulatory T-cells have at least one characteristic selected from the group consisting of being CD4 positive, being CD25^hi, and being CD127⁻

.

12. The method of claim 11, wherein at least some of the initial regulatory T-cells have at least one characteristic selected from the group consisting of being Foxp3 positive and being Helios positive.

13. The method of claim 1, further comprising contacting the population of cells with at least one molecule selected from the group consisting of an anti-CDRε

antibody, an anti-CD28 antibody, and interleukin-2.

14. The method of claim 1, wherein expansion of the initial regulatory T-cells results in an expanded population of cells in which at least 60% of the cells are stable, regulatory T-cells.

18. A composition comprising a population of stable, regulatory T cells produced according to the method of claim 1.

15. A method for stabilizing T regulatory cells, the method comprising culturing isolated cells comprising initial regulatory T-cells, in the presence of an oligonucleotide to expand the initial regulatory T-cell population.

16. A composition comprising isolated cells, wherein greater than 60% of the isolated cells are stable, regulatory T-cells.

17. The composition of claim 16, wherein the stable regulatory T-cells are Foxp3+Helios+.

19. A method of treating an individual for an autoimmune disease comprising administering the composition of claim 16 or 18 to the individual.