Gamma delta T cells and a method of augmenting the tumoricidal activity of the same

Patent metadata

Field	Value
Publication number	US-11629333-B2
Application number	US-201816484427-A
Country	US
Kind code	B2
Filing date	Feb 8, 2018
Priority date	Feb 8, 2017
Publication date	Apr 18, 2023
Grant date	Apr 18, 2023

Abstract

The present invention relates to a method of generating γδ T cells having at least one down-regulated co-inhibitory receptor, the method comprising the steps of: (a) culturing a population of cells comprising γδ T cells with a phosphoantigen to expand the γδ T cells; and (b) culturing the expanded γδ T cells with artificial antigen-presenting cells expressing a Fc receptor, and an anti-CD3 antibody. The present invention also relates to γδ T cells generated according to a method of the present invention, as well as methods of treatment and medical uses thereof.

First claim

The invention claimed is:

1. A method of generating γδ T cells having at least one down-regulated co-inhibitory receptor, the method comprising the steps of:
(a) culturing a population of cells comprising Vγ9Vδ2 T cells with a phosphoantigen to expand the Vγ9Vδ2 T cells;
(b) culturing the expanded Vy9V62T cells with artificial antigen-presenting cells expressing a Fc receptor, and an anti-CD3 antibody; and
(c) modifying the Vγ9Vδ2 T cells to express a chimeric antigen receptor (CAR) wherein the CAR overexpresses an extracellular antigen binding domain of natural killer group 2D (NKG2D).

2. The method of claim 1 , wherein the Fc receptor is CD64.

3. The method of claim 1 , wherein the phosphoantigen is zoledronic acid or a salt thereof.

4. The method of claim 1 , wherein the population of cells are peripheral blood mononuclear cells.

5. The method of claim 1 , wherein step (a) is carried out for 7 days.

6. The method of claim 1 , wherein step (b) is carried out for 10 days or more.

7. The method of claim 1 , wherein the artificial antigen-presenting cells are K562 cells.

8. The method of claim 1 , further comprising irradiating the artificial antigen-presenting cells prior to step (b) using gamma irradiation.

9. The method of claim 1 , wherein the anti-CD3 antibody is Muromonab-CD3.

10. The method of claim 1 , wherein the at least one co-inhibitory receptor is selected from the group consisting of cytotoxic T lymphocyte (CTL)-associated antigen 4 (CTLA-4/CD152); programmed cell death protein 1 (PD-1/CD279); lymphocyte activation gene-3 (LAG-3); T cell immunoglobulin and immunoreceptor tyrosine-based inhibition motif (ITIM) domain (TIGIT); T-cell immunoglobulin and mucin-containing protein 3 (TIM3); and B and T lymphocyte attenuator (BTLA).

11. The method of claim 1 , wherein the modified Vγ9Vδ2 T cells have at least one up-regulated activating receptor.

12. The method of claim 1 , wherein modifying the Vγ9Vδ2 T cells comprises transfecting the Vγ9Vδ2 T cells with an mRNA vector encoding the CAR.

13. The method of claim 12 , wherein transfecting the Vγ9Vδ2 T cells comprises RNA electroporation.

14. The method of claim 1 , wherein the CAR further comprises a signalling domain of CD3 zeta or DAP 12.

Assignees

• Agency Science Tech & Res

Inventors

• Wang Shu
• Ang Wei Xia
• Zeng Jieming

Classifications

• A61K40/4224

  Molecules with a "CD" designation not provided for elsewhere · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/38

  characterised by the dose, timing or administration schedule · CPC title

• A61K2239/59

  Reproductive system, e.g. uterus, ovaries, cervix or testes · CPC title