T Cell Expansion Method
The invention relates to the expansion of T cells and particularly, although not exclusively, to the expansion of gamma delta T cells, and the optimization of medium, serum and cytokine combinations for large-scale ex vivo expansion of gamma delta T cells for clinical use.
1 . A method for generating or expanding gamma delta T cells, the method comprising culturing peripheral blood mononuclear cells (PBMCs) in the presence of IL2 and IL21.
2 . A method for generating or expanding gamma delta T cells, the method comprising culturing PBMCs in the presence of 112 and IL18.
3 . A method for generating or expanding gamma delta T cells, the method comprising culturing PBMCs in the presence of IL15.
4 . The method according to claim 3 , wherein the PBMCs are cultured in the presence of IL15 and IL21.
5 . The method according to claim 4 , wherein the PBMCs are cultured in the presence of IL15, IL 21 and IL18.
6 . A method for generating or expanding gamma delta T cells, the method comprising culturing PBMCs in the presence of IL21.
7 . The method according to claim 6 wherein the PBMCs are cultured in the presence of IL21 and IL2 and/or IL15.
8 . The method according to any one of the preceding claims wherein the gamma delta T cells are Vδ2 T cells.
9 . The method according to claim 8 , wherein the gamma delta T cells are Vγ9Vδ2 T cells.
10 . The method according to any one of the preceding claims, wherein the method comprises culturing the PBMCs in culture medium supplemented with serum.
11 . The method according to claim 10 wherein the culture medium is supplemented with 10% serum.
12 . The method according to any one of the preceding claims the PBMCs are cultured in OpTimizer® T cell media.
13 . The method according to claim 10 or claim 11 wherein the serum is human AB serum or defined FBS.
14 . The method according to any one of claims any one of the preceding claims, wherein the method generates a population of gamma delta T cells that is at least 60% gamma delta T cells, preferably at least 70% gamma delta T cells.
15 . The method according to any one of the preceding claims, wherein the gamma delta T cells exhibit antigen presentation and effector phenotypes.
16 . A gamma delta T cell generated using a method according to any one of the preceding claims.
17 . The gamma delta T cell according to claim 16 for use in medicine.
18 . The gamma delta T cell according to claim 16 for use in a method of adoptive T cell therapy.
19 . A gamma delta T cell that expresses a higher level of at least one marker selected from HLA-ABC, HLA-DR, CD80, CD83, CD86, CD40 and ICAM-1 than a gamma delta T cell that has been generated in the presence of IL2 alone.
20 . A gamma delta T cell that expresses a higher level of at least one marker selected from CCR5, CCR6, CCR7, CD27 and NKG2D than a gamma delta T cell that has been generated in the presence of IL2 alone.
21 . A cell culture comprising gamma delta T cells, media, and
IL2 and IL21;
IL15;
IL15 and IL21;
IL2 and IL18;
IL15, IL18 and IL21.
IL2 and IL7;
IL2 and IL15;
IL2, IL18 and IL21;
IL15 and IL7; or
IL15 and IL18.
22 . The cell culture according to claim 21 , further comprising serum, preferably 10% serum.
23 . A method for generating or expanding a population of antigen-specific T cells, comprising stimulating T cells by culture in the presence of gamma delta T cells generated/expanded according to the method of any one of claims 1 to 15 presenting a peptide of the antigen.
24 . An antigen-specific T cell according generated using the method according to claim 23 .
25 . The antigen-specific T cell according to claim 24 for use in medicine.
26 . The antigen-specific T cell according to claim 25 for use in a method of adoptive T cell therapy.
27 . A method of treating or preventing a disease or disorder in a subject, comprising:
(a) isolating PBMCs from a subject;
(b) generating or expanding a population of gamma delta T cells according to the method of any one of claims 1 to 15 , and;
(c) administering the gamma delta T cells to a subject.
28 . A method of treating or preventing a disease or disorder in a subject, comprising:
(a) isolating PBMCs from a subject;
(b) generating or expanding a population of gamma delta T cells according to the method of any one of claims 1 to 15 ;
(c) generating or expanding a population of antigen-specific T cells by a method comprising stimulating T cells by culture in the presence of gamma delta T cells generated/expanded according to (b) presenting a peptide of the antigen; and
(d) administering the antigen-specific T cells to a subject.