Anti third party central memory t cells, methods of producing same and use of same in transplantation and disease treatment

Abstract

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)