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 generating an isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, said cells being tolerance-inducing cells and/or endowed with anti-disease activity, and capable of homing to the lymph nodes following transplantation, 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 said 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 said central memory T-lymphocyte (Tcm) phenotype, thereby generating the isolated population of cells.
2 . The method of claim 1 , further comprising:
depleting non-adherent cells from said PBMC prior to step (a);
depleting CD4+ and/or CD56+ cells from said PBMC prior to step (a); and/or
selecting CD45RA+ and/or CD45RO− cells from said PBMC prior to step (a).
3 - 5 . (canceled)
6 . The method of claim 1 , further comprising culturing said cells resulting from step (a) with a third party antigen or antigens in the presence of IL-21, IL-15 and IL-7 following step (a) and prior to step (b).
7 - 10 . (canceled)
11 . A method of generating an isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, said cells being tolerance-inducing cells and/or endowed with graft-versus-leukemia (GVL) activity, and capable of homing to the lymph nodes following transplantation, the method comprising:
(a) treating non-adherent peripheral blood mononuclear cells (PBMC) with an agent capable of depleting CD4+ and/or CD56+ cells so as to obtain CD8+ T cells;
(b) contacting said CD8+ T cells with third party dendritic cells in the presence of IL-21 for 12 hours to 5 days so as to allow enrichment of antigen reactive cells;
(c) culturing said cells resulting from step (b) with said third party dendritic cells in the presence of IL-21, IL-15 and IL-7 for 12 hours to 3 days; 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 for 5-20 days so as to allow proliferation of cells comprising said central memory T-lymphocyte (Tcm) phenotype, thereby generating the isolated population of cells.
12 . A method of generating an isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, said cells being endowed with anti-disease activity, and capable of homing to the lymph nodes following transplantation, the method comprising:
(a) treating non-adherent peripheral blood mononuclear cells (PBMC) with an agent capable of depleting CD4+ and/or CD56+ cells so as to obtain CD8+ T cells;
(b) contacting said CD8+ T cells with non-syngeneic dendritic cells in the presence of IL-21 for 12 hours to 5 days so as to allow enrichment of antigen reactive cells;
(c) culturing said cells resulting from step (b) with said non-syngeneic dendritic cells in the presence of IL-21, IL-15 and IL-7 for 12 hours to 3 days; 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 for 5-20 days so as to allow proliferation of cells comprising said central memory T-lymphocyte (Tcm) phenotype, thereby generating the isolated population of cells.
13 . The method of claim 12 , further comprising selecting CD45RA+ and/or CD45RO− cells from said PBMC following step (a) and prior to step (b).
14 - 16 . (canceled)
17 . The method of claim 1 , wherein said contacting in said presence of IL-21 is effected for 12 hours to 5 days.
18 - 19 . (canceled)
20 . The method of claim 1 , further comprising:
selecting for activated cells following step (a) and prior to step (b); or
depleting alloreactive cells following step (b).
21 . The method of claim 11 , further comprising:
selecting CD45RA+ and/or CD45RO− cells from said PBMC following step (a) and prior to step (b); or
selecting for activated cells following step (b) and prior to step (c); or
depleting alloreactive cells following step (d).
22 - 24 . (canceled)
25 . The method of claim 1 , wherein said culturing in said presence of IL-21, IL-15 and IL-7 in said antigen free environment is effected for 5-20 days.
26 - 32 . (canceled)
33 . The method of claim 1 , wherein said anti-third party cells having a T central memory phenotype comprises a CD3 + , CD8 + , CD62L + , CD45RA − , CD45RO + signature.
34 . The method of claim 33 , wherein at least 50% of the isolated population of cells are CD3+CD8+ cells of which at least 50% have said signature.
35 . An isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, wherein at least 50% of the isolated population of cells are CD3+CD8+ cells of which at least 50% comprise a CD3 + , CD8 + , CD62L + , CD45RA − , CD45RO + signature, and further wherein said cells are tolerance-inducing cells and/or endowed with anti-disease activity, and capable of homing to the lymph nodes following transplantation.
36 . An isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, said cells being tolerance-inducing cells and/or endowed with anti-disease activity, and capable of homing to the lymph nodes following transplantation, generated according to the method of claim 1 .
37 . A method of treating a disease in a subject in need thereof, wherein the disease is selected from the group consisting of a malignant disease, a viral disease and an autoimmune disease, the method comprising administering to the subject a therapeutically effective amount of the isolated population of cells of claim 36 , thereby treating the subject.
38 - 40 . (canceled)
41 . A method of treating a subject in need of a cell or tissue transplantation, the method comprising:
(a) transplanting a cell or tissue transplant into the subject; and
(b) administering to the subject a therapeutically effective amount of the isolated population of cells of claim 36 , thereby treating the subject.
42 - 44 . (canceled)
45 . The method of claim 41 , wherein said cell or tissue transplant comprises immature hematopoietic cells.
46 . The method of claim 41 , wherein said cell or tissue transplant is selected from the group consisting of a liver, a pancreas, a spleen, a kidney, a heart, a lung, a skin, an intestine and a lymphoid/hematopoietic tissue or organ.
47 . The method of claim 41 , wherein said cell or tissue transplant comprises a co-transplantation of several organs.
48 . The method of claim 47 , wherein said co-transplantation comprises transplantation of immature hematopoietic cells and a solid organ.
49 . The method of claim 48 , wherein said immature hematopoietic cells and said solid organ are obtained from the same donor.
50 . The method of claim 48 , wherein said immature hematopoietic cells are transplanted prior to, concomitantly with, or following said transplantation of said solid organ.
51 . The method of claim 41 , wherein said isolated population of cells are administered prior to, concomitantly with, or following said cell or tissue transplant.
52 - 53 . (canceled)
54 . The method of claim 41 , wherein said cell or tissue transplant and said isolated population of cells are derived from the same donor.
55 . The method of claim 41 , wherein said cell or tissue transplant is syngeneic with the subject and said isolated population of cells are non-syngeneic with the subject.
56 . The method of claim 41 , wherein said cell or tissue transplant is syngeneic with the subject and said isolated population of cells are syngeneic with the subject.
57 . A method of treating a subject in need of an immature hematopoietic cell transplantation, the method comprising:
(a) transplanting immature hematopoietic cells into the subject; and
(b) administering to the subject a therapeutically effective amount of the isolated population of cells of claim 36 , thereby treating the subject.
58 . The method of claim 57 , wherein said isolated population of cells are administered prior to, concomitantly with, or following said immature hematopoietic cells.
59 . The method of claim 57 , wherein said immature hematopoietic cells and said isolated population of cells are derived from the same donor.
60 . (canceled)
61 . The method of claim 57 , wherein said immature hematopoietic cells and said isolated population of cells are derived from the subject.
62 - 63 . (canceled)