Process of expanding T cells
The present disclosure relates to a novel process for expanding T cells, such as autologous T cells, cell populations therefrom, pharmaceutical compositions comprising the said cell populations and use of the cells and compositions for treatment, particular the treatment or prophylaxis of virus infection and/or cancer, for example in immune compromised or immune competent human patients.
1. A process for in vitro expansion of autologous antigen specific T cells comprising the steps:
a) culturing a population of autologous PBMC cells in the presence of:
i) dendritic cells which have been pulsed with a peptide/peptide mix relevant to a target antigen(s) OR a peptide/peptide mix relevant to a target antigen(s), and
ii) at least one cytokine, and
b) culturing a population of T cells from step a) in the presence of:
i) autologous antigen presenting T cells (T-APCs) which have been pulsed with a peptide/peptide mix relevant to a target antigen(s), and
ii) an artificial co-stimulatory favor,
characterized in that the process does not employ live virus and/or viral vectors or the use of DNA or RNA encoding antigens in the expansion of the relevant T cell population.
2. A process according to claim 1 , wherein step b) further comprises population of T cells from step a) in the presence of a cytokine.
3. A process according to claim 1 wherein step b) is performed two or more times until sufficient quantities of the relevant T cell population are obtained.
4. A process according to claim 1 wherein the culturing of step a) is performed for 12 days or less.
5. A process according to claim 1 , wherein the culturing step of step b) is performed for 12 days or less.
6. A process according to claim 1 , wherein culturing is performed in a vessel comprising a gas permeable culture surface.
7. A process according to claim 1 , wherein the T cells expanded are specific to a viral antigen or antigens of Epstein-Barr Virus, Vaccinia Virus or Varicella Zoster Virus.
8. A process according to claim 1 , wherein the peptides of step a) and/or b) comprise between 2 and 1000 peptides.
9. A process according to claim 1 , wherein the peptides of step a) and/or b) overlap by 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more amino acids.
10. A process according to claim 1 , wherein the peptides of step a) and/or b) are about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length.
11. A process according to claim 1 , wherein the peptides of part a) and/or b) cover part or the full length of the antigen LMPI.
12. A process according to claim 1 , wherein the peptides of part a) and/or b) cover part or the full length of the antigen LMP2.
13. A process according to claim 1 , wherein the peptides of part a) and/or b) cover part of or the full length of the antigen EBNA1.
14. A process according to claim 1 , wherein the peptides of part a) and/or b) cover part of or the full length of the antigen BARF1.
15. A process according to claim 1 , wherein the cytokine present in step a) is IL-4 and/or IL-7.
16. A process according to claim 2 , wherein the cytokine present in step b) is IL-15.
17. A process according to claim 1 , wherein the artificial co-stimulatory factor is an engineered cell line with one or more relevant protein or protein fragments present on the cell surface.
18. A process according to claim 17 , wherein the protein or protein fragments are independently selected from CD80, CD86, CD83, OX-40 ligand and 41BB-ligand.
19. A process according to claim 18 , wherein all of the said protein or protein fragments are present on the surface of the co-stimulatory cell.
20. A process according to claim 1 , wherein the process is performed in a GRex™ system.
21. A process according to claim 17 , wherein the artificial co-stimulatory factor is a HLA negative cell line which has been genetically modified to express co-stimulatory molecules.
22. A process according to claim 17 , wherein the artificial co-stimulatory factor is an engineered aK562 cell.
23. A process according to claim 1 , wherein the artificial co-stimulatory factor is a bead with one or more relevant protein or protein fragments present on the cell surface.
24. A process according to claim 22 , wherein the protein or protein fragments are independently selected from CD80, CD86, anti-CD28 and anti-4-1BB.
25. A process according to claim 21 , wherein an anti-CD3 antibody is not loaded onto the Fcy receptor on the surface of the engineered cell.
26. A process according to claim 1 , wherein the population of autologous PBMC cells is cultured in the presence of the dendritic cells which have been pulsed with the peptide/peptide mix relevant to the target antigen(s).
27. A process according to claim 1 , wherein the population of autologous PBMC cells is cultured in the presence of the peptide/peptide mix relevant to the target antigen(s).
28. A process according to claim 1 , wherein the peptides of step a) comprise between 2 and 1000 peptides.
29. A process according to claim 1 , wherein the peptides of step b) comprise between 2 and 1000 peptides.
30. A process according to claim 1 , wherein the peptides of step a) overlap by 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more amino acids.
31. A process according to claim 1 , wherein the peptides of step b) overlap by 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more amino acids.
32. A process according to claim 1 , wherein the peptides of step a) are about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length.
33. A process according to claim 1 , wherein the peptides of step b) are about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length.
34. A process according to claim 1 , wherein the peptides of part a) cover part or the full length of the antigen LMPI.
35. A process according to claim 1 , wherein the peptides of part b) cover part or the full length of the antigen LMPI.
36. A process according to claim 1 , wherein the peptides of part a) cover part or the full length of the antigen LMP2.
37. A process according to claim 1 , wherein the peptides of part b) cover part or the full length of the antigen LMP2.
38. A process according to claim 1 , wherein the peptides of part a) cover part of or the full length of the antigen EBNA1.
39. A process according to claim 1 , wherein the peptides of part b) cover part of or the full length of the antigen EBNA1.
40. A process according to claim 1 , wherein the peptides of part a) cover part of or the full length of the antigen BARF1.
41. A process according to claim 1 , wherein the peptides of part b) cover part of or the full length of the antigen BARF1.
42. A process according to claim 14 , wherein the cytokine present in step a) is IL-4.
43. A process according to claim 14 , wherein the cytokine present in step a) is IL-7.
44. A process according to claim 14 , wherein the cytokine present in step a) is IL-4 and IL-7.