Anti third party central memory t cells, methods of producing same and use of same in transplantation and disease treatment
A method of generating an isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, the cells being tolerance-inducing cells and/or endowed with anti-disease activity, and capable of homing to the lymph nodes following transplantation is disclosed. The method comprising: (a) contacting peripheral blood mononuclear cells (PBMC) with a third party antigen or antigens in the presence of IL-21 so as to allow enrichment of antigen reactive cells; and (b) culturing the cells resulting from step (a) in the presence of IL-21, IL-15 and IL-7 in an antigen free environment so as to allow proliferation of cells comprising the central memory T-lymphocyte (Tcm) phenotype.
1. A method of treating an autoimmune disease in a subject in need thereof, the method comprising:
(i) generating an isolated population of cells comprising non-graft versus host (GVH) inducing anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, said cells being tolerance-inducing cells and capable of homing to the lymph nodes following administration to the subject, by a method comprising:
(a) treating peripheral blood mononuclear cells (PBMC) derived from a donor with an agent capable of depleting CD4+ and CD56+ cells so as to obtain CD8+ T cells;
(b) contacting said CD8+ T cells with a third-party antigen or antigens in the presence of IL-21 so as to allow enrichment of antigen reactive cells;
(c) adding IL-15 and IL-7 to said culture comprising said antigen reactive cells, said third party antigen or antigens and said IL-21 of step (b); and
(d) culturing said cells resulting from step (c) in the presence of IL-21, IL-15 and IL-7 in an antigen free environment so as to allow proliferation of cells comprising said Tcm phenotype;
(ii) transplanting into the subject immature hematopoietic cells derived from said donor; and
(iii) administering to the subject a therapeutically effective amount of said isolated population of cells,
thereby treating the subject.
2. The method of claim 1 , wherein said isolated population of cells are syngeneic with the subject.
3. The method of claim 1 , wherein said isolated population of cells are non-syngeneic with the subject.
4. The method of claim 1 , wherein said subject is a human subject.
5. The method of claim 1 , wherein said third party antigen or antigens are presented on antigen presenting cells.
6. The method of claim 5 , wherein said antigen presenting cells comprise dendritic cells.
7. The method of claim 6 , wherein said dendritic cells comprise irradiated dendritic cells.
8. The method of claim 6 , wherein said dendritic cells comprise third party antigen or antigens pulsed dendritic cells.
9. The method of claim 1 , wherein said third party antigen or antigens are selected from the group consisting of a viral antigen, a bacterial antigen, a protein extract, a purified protein and a synthetic peptide.
10. The method of claim 1 , further comprising:
depleting adherent cells from said PBMC prior to step (a); and/or
selecting CD45RA + and/or CD45RO − cells from said PBMC prior to step (a).
11. The method of claim 1 , wherein said contacting in said presence of IL-21 according to step (b) is affected for 12 hours to 5 days.
12. The method of claim 1 , wherein said contacting in said presence of IL-21 according to step (b) is affected for 2-3 days.
13. The method of claim 1 , wherein step (c) is affected for 12 hours to 3 days.
14. The method of claim 1 , wherein step (c) is affected for 1-2 days.
15. The method of claim 1 , wherein step (d) is affected for 5-20 days.
16. The method of claim 1 , wherein step (d) is affected for 7-11 days.
17. The method of claim 1 , further comprising:
selecting for activated cells following step (b) and prior to step (d); or
depleting alloreactive cells following step (d).
18. The method of claim 1 , wherein said anti-third party cells having a Tcm phenotype comprises a CD3 + , CD8 + , CD62L + , CD45RA-CD45RO + signature.
19. The method of claim 18 , wherein at least 50% of the isolated population of cells are CD3+CD8+ cells of which at least 50% have said signature.