METHODS OF T CELL PRODUCTION

1 . A method of producing a population of TCR αβ+ T cells comprising;

(i) differentiating a population of haematopoietic progenitor cells (HPCs) into progenitor T cells and;

(ii) maturing the progenitor T cells to produce a population of TCR αβ+ T cells,

wherein one or both of (i) and (ii) are performed in the presence of Inducible Co-stimulator ligand (ICOS-L).

2 . A method according to claim 1 wherein the presence of ICOS-L increases the proportion of HPCs or progenitor T cells that mature into TCR αβ+ cells.

3 . A method according to claim 1 or claim 2 wherein the HPCs are differentiated into progenitor T cells in the presence of ICOS-L.

4 . A method according to any one of claims 1 to 3 wherein the progenitor T cells are matured in the presence of ICOS-L.

5 . A method according to any one of the preceding claims wherein the HPCs and/or progenitor T cells are cultured on a surface coated with ICOS-L.

6 . A method according to any one of the preceding claims wherein the HPCs have a CD34+ phenotype.

7 . A method according to any one of the preceding claims wherein the population of HPCs is produced in vitro from induced pluripotent stem cells (iPSCs).

8 . A method according to claim 7 wherein the method comprises providing a population of iPSCs and differentiating the iPSCs into a population of HPCs.

9 . A method according to claim 7 or claim 8 wherein the iPSCs are derived from T cells obtained from a donor individual.

10 . A method according to claim 9 wherein the T cells obtained from the donor individual are specific for a target antigen.

11 . A method according to claim 10 wherein the target antigen is a tumour antigen.

12 . A method according to claim 10 or 11 wherein the T cells obtained from the donor individual are tumour-infiltrating lymphocytes (TILs).

13 . A method according to any one of the preceding claims wherein the HPCs are differentiated by a method comprising culturing the population of HPCs in a lymphoid expansion medium to produce the progenitor T cells.

14 . A method according to any one of the preceding claims wherein the progenitor T cells have a CD5+, CD7+ phenotype.

15 . A method according to any one of the preceding claims wherein the progenitor T cells are differentiated by a method comprising culturing the population of progenitor T cells in a T cell maturation medium to produce the TCR αβ+ T cells.

16 . A method according to any one of the preceding claims wherein the TCR αβ+ T cells have a CD8+CD4+ phenotype.

17 . A method according to any one of the preceding claims comprising activating and expanding the TCR αβ+ T cells to produce a population of T cells have a CD8+ single positive phenotype or a CD4+ single positive phenotype.

18 . A method according to any one of the preceding claims wherein the TCR αβ+ T cells specifically bind to cells expressing a target antigen.

19 . A method according to claim 18 wherein the target antigen is a tumour antigen.

20 . A method according to claim 19 wherein the TCR αβ+ T cells specifically bind to cancer cells expressing the tumour antigen.

21 . A method according to any one of claims 1 to 20 wherein the method further comprises introducing heterologous nucleic acid encoding an αβ TCR into the iPSCs, HPCs or progenitor T cells.

22 . A method according to claim 21 wherein the heterologous nucleic acid encoding the αβ TCR is comprised in an expression vector.

23 . A method according to claim 22 wherein the expression vector is a lentiviral vector.

24 . A method according to any one of claims 21 to 23 wherein the αβ TCR is an affinity enhanced TCR.

25 . A method according to one of claims 21 to 24 wherein the αβ TCR binds specifically to an MHC displaying a peptide fragment of a target antigen expressed by cells or specifically binds to a target antigen or peptide thereof expressed by cells independently of MHC presentation.

26 . A method according to claim 25 wherein the αβ TCR binds specifically to an MHC displaying a peptide fragment of a tumour antigen expressed by the cancer cells or binds specifically to a tumour antigen or peptide fragment thereof expressed by cancer cells independently of MHC presentation.

27 . A method according to any one of the preceding claims further comprising isolating or purifying the TCR αβ+ T cells.

28 . A method according to claim 27 wherein TCR αβ+ T cells are isolated by magnetic activated cell sorting.

29 . A method according to according to any one of the preceding claims comprising concentrating the population of TCR αβ+ T cells.

30 . A method according to according to any one of the preceding claims comprising storing the population of TCR αβ+ T cells.

31 . A method according to any one of the preceding claims comprising formulating the population of TCR αβ+ T cells with a pharmaceutically acceptable excipient.

32 . A population of TCR αβ+ T cells produced by a method according to any one of claims 1 to 31 .

33 . A pharmaceutical composition comprising a population of TCR αβ+ T cells produced by a method according to any one of claims 1 to 31 and a pharmaceutically acceptable excipient.

34 . A population of TCR αβ+ T cells produced by a method according to any one of claims 1 to 31 for use in a method of treatment.

35 . A population of TCR αβ+ T cells produced by a method according to any one of claims 1 to 31 for use in a method of treatment of cancer.

36 . A coating composition for treating the surface of a cell culture vessel, the composition comprising ICOS-L.

37 . A composition according to claim 36 further comprising a Notch signalling ligand.

38 . A culture vessel for T cell culture, said vessel comprising a surface coated with ICOS-L.

39 . A culture vessel according to claim 39 wherein the surface is additionally coated with a Notch signalling ligand.