Veto cells effective in preventing graft rejection and devoid of graft versus host potential

Abstract

A method of transplanting a transplant derived from a donor into a recipient is disclosed. The method comprises the steps of (a) transplanting the transplant into the recipient; and (b) administering to the recipient a dose including non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), wherein the non-alloreactive anti-third party CTLs are generated by directing T-lymphocytes of the donor against a third party antigen or antigens, the dose is substantially depleted of T-lymphocytes capable of developing into alloreactive CTLs, thereby preventing or ameliorating both graft rejection by the recipient and graft versus host disease.

1 . A method of preventing or decreasing graft rejection in a human recipient of a transplant which is derived from a non-syngeneic donor, the method comprising: 
 (a) conditioning the recipient under lethal or sublethal conditions;    (b) transplanting the transplant into the recipient; and    (c) administering to the recipient a dose of a cell preparation, wherein said cell preparation comprises non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), does not comprise lymphocytes capable of developing into anti-recipient CTLs, and does not comprise CD4+ cells and/or CD56+ cells, and wherein said cell preparation is generated by a method which comprises: 
 (i) directing peripheral blood lymphocytes derived from the non-syngeneic donor against a third party antigen or antigens, thereby generating third party antigen-treated lymphocytes which comprise anti-third party CTLs;  
 (ii) depleting said peripheral blood lymphocytes derived from the non-syngenic donor of CD4+ cells and/or CD56+ cells; and  
 (iii) depleting, by affinity purification, said third party antigen-treated lymphocytes of T-lymphocytes capable of developing into anti-recipient CTLs, thereby preventing or decreasing graft rejection in the recipient of the transplant.  
   
   
   
       2 . The method of  claim 1 , wherein the non-syngeneic donor is allogeneic with respect to the recipient.  
   
   
       3 . The method of  claim 1 , wherein the non-syngeneic donor is xenogeneic with respect to the recipient.  
   
   
       4 . The method of  claim 1 , wherein the non-syngeneic donor is HLA identical with respect to the recipient.  
   
   
       5 . The method of  claim 1 , wherein the non-syngeneic donor is HLA non-identical with respect to the recipient.  
   
   
       6 . The method of  claim 1 , wherein the non-syngeneic donor is mismatched haploidentical with respect to the recipient.  
   
   
       7 . The method of  claim 1 , wherein the transplant is selected from the group consisting of cells, a tissue and an organ.  
   
   
       8 . The method of  claim 1 , wherein said third party antigen or antigens is selected from the group consisting of third party cells, a cell antigen, a viral antigen, a bacterial antigen, a protein extract, a purified protein, a synthetic peptide presented by autologous antigen presenting cells and a synthetic peptide presented by non-autologous antigen presenting cells.  
   
   
       9 . The method of  claim 8 , wherein said third party cells are allogeneic or xenogeneic with respect to the recipient.  
   
   
       10 . The method of  claim 9 , wherein said third party cells have HLA antigens which are different from HLA antigens of the non-syngeneic donor but which are not cross reactive with HLA antigens of the recipient.  
   
   
       11 . The method of  claim 9 , wherein said third party cells are stimulatory cells selected from the group consisting of cells purified from peripheral blood lymphocytes, cells purified from spleen, cells purified from lymph nodes, cytokine-mobilized PBLs and in vitro expanded antigen-presenting cells (APC).  
   
   
       12 . The method of  claim 1 , wherein steps (a) and (b) are effected at the same time.  
   
   
       13 . The method of  claim 1 , wherein step (a) is effected prior to step (b).  
   
   
       14 . The method of  claim 1 , wherein step (a) is effected following step (b).  
   
   
       15 . The method of  claim 1 , wherein affinity purification is effected by an antibody.  
   
   
       16 . The method of  claim 15 , wherein the antibody is selected from the group consisting of anti-CD69, anti-CD25, anti-IL-2 and anti-INFγ.  
   
   
       17 . The method of  claim 1 , wherein the transplant comprises immature hematopoietic cells.  
   
   
       18 . The method of  claim 17 , wherein said immature hematopoietic cells are CD34+ cells.  
   
   
       19 . The method of  claim 17 , wherein said immature hematopoietic cells are stem cells.  
   
   
       20 . The method of  claim 1 , wherein the recipient has a malignant disease.  
   
   
       21 . The method of  claim 1 , wherein the recipient has a hematopoietic malignant disease.  
   
   
       22 . The method of  claim 1 , wherein the recipient has a lymphoma.  
   
   
       23 . A method of treating a human recipient suffering from a disease requiring immature hematopoietic cell transplantation, the method comprising: 
 (a) conditioning the recipient under lethal or sublethal conditions;    (b) administering to the recipient a dose which comprises immature hematopoietic cells derived from a non-syngeneic donor which is allogeneic with respect to the recipient; and    (c) administering to the recipient a dose of a cell preparation which comprises non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), does not comprise T-lymphocytes capable of developing into anti-recipient CTLs, does not comprise CD4+ cells and/or CD56+ cells, and is generated by a method which comprises: 
 (i) directing peripheral blood lymphocytes derived from the non-syngeneic donor against a third party antigen or antigens, thereby generating third party antigen-treated lymphocytes which comprise anti-third party CTLs.  
 (ii) depleting said peripheral blood lymphocytes derived from said non-syngeneic donor of CD4+ cells and/or CD56+ cells; and  
 (iii) depleting, by affinity purification, said third party antigen-treated lymphocytes of T-lymphocytes capable of developing into anti-recipient CTLs, thereby preventing or treating the recipient suffering from the disease requiring immature hematopoietic cell transplantation.  
   
   
   
       24 . The method of  claim 23 , wherein the non-syngeneic donor is allogeneic with respect to the recipient.  
   
   
       25 . The method of  claim 23 , wherein the non-syngeneic donor is xenogeneic with respect to the recipient.  
   
   
       26 . The method-of  claim 23 , wherein said non-syngeneic donor is HLA identical with respect to the recipient.  
   
   
       27 . The method-of  claim 23 , wherein said non-syngeneic donor is HLA non-identical with respect to the recipient.  
   
   
       28 . The method-of  claim 23 , wherein said non-syngeneic donor is mismatched haploidentical with respect to the recipient.  
   
   
       29 . The method of  claim 23 , wherein said third party antigen or antigens is selected from the group consisting of third party cells, a cell antigen, a viral antigen, a bacterial antigen, a protein extract, a purified protein, a synthetic peptide presented by autologous antigen presenting cells and a synthetic peptide presented by non-autologous antigen presenting cells.  
   
   
       30 . The method of  claim 29 , wherein said third party cells are allogeneic or xenogeneic with respect to the recipient.  
   
   
       31 . The method of  claim 30 , wherein said third party cells have HLA antigens which are different from HLA antigens of said non-syngeneic donor and which are not cross reactive with HLA antigens of the recipient.  
   
   
       32 . The method of  claim 30 , wherein said third party cells are stimulatory cells selected from the group consisting of cells purified from peripheral blood lymphocytes, cells purified from spleen, cells purified from lymph nodes, cytokine-mobilized PBLs and in vitro expanded antigen-presenting cells (APC).  
   
   
       33 . The method of  claim 23 , wherein said immature hematopoietic cells are cells which are derived from a source selected from the group consisting of bone marrow, mobilized peripheral blood, fetal liver, yolk sac and/or cord blood.  
   
   
       34 . The method of  claim 33 , wherein said cells which are derived from mobilized peripheral blood are obtained by leukapheresis of peripheral blood of said non-syngeneic donor after stimulation of said non-syngeneic donor with a suitable cytokine.  
   
   
       35 . The method of  claim 23 , wherein affinity purification is effected by an antibody.  
   
   
       36 . The method of  claim 35 , wherein the antibody is selected from the group consisting of anti-CD69, anti-CD25, anti-IL-2 and anti-INFγ.  
   
   
       37 . The method of  claim 23 , wherein said immature hematopoietic cells are T-cell depleted hematopoietic progenitor cells.  
   
   
       38 . The method of  claim 23 , wherein said immature hematopoietic cells are CD34+ cells.  
   
   
       39 . The method of  claim 23 , wherein a cell ratio between said cytotoxic T-lymphocytes and said immature hematopoietic cells is at least 1 to 100.  
   
   
       40 . The method of  claim 23 , wherein said immature hematopoietic cells are stem cells.  
   
   
       41 . The method of  claim 23 , wherein steps (b) and (c) are effected at the same time.  
   
   
       42 . The method of  claim 23 , wherein step (b) is effected following step (c).  
   
   
       43 . The method of  claim 23 , wherein the disease is a malignant disease.  
   
   
       44 . The method of  claim 23 , wherein the disease is a hematopoietic malignancy.  
   
   
       45 . The method of  claim 23 , wherein the disease is a lymphoma.  
   
   
       46 . A method of producing a cell preparation for preventing or decreasing graft rejection in a human recipient of a transplant, wherein the recipient is conditioned under sublethal conditions, wherein the transplant is derived from a non-syngeneic donor, wherein the cell preparation comprises non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), wherein the cell preparation does not comprise T-lymphocytes capable of developing into anti-recipient CTLs, and wherein the cell preparation does not comprise CD4+ cells and/or CD56+ cells, the method comprising: 
 (i) directing peripheral blood lymphocytes derived from the non-syngeneic donor against a third party antigen or antigens, thereby generating third party antigen-treated lymphocytes which comprise anti-third party CTLs;    (ii) depleting, by affinity purification, said third party antigen-treated lymphocytes of T-lymphocytes capable of developing into anti-recipient CTLs; and    (iii) removing CD4+ cells and/or CD56+ cells from said peripheral blood lymphocytes derived from the non-syngeneic donor, thereby producing the cell preparation.    
   
   
       47 . The method of  claim 46 , wherein the non-syngeneic donor is allogeneic with respect to the recipient.  
   
   
       48 . The method of  claim 46 , wherein the non-syngeneic donor is xenogeneic with respect to the recipient.  
   
   
       49 . The method of  claim 46 , wherein step (iii) is effected by removing both the CD56+ cells and the CD4+ cells from said peripheral blood lymphocytes derived from the non-syngeneic donor.  
   
   
       50 . The method of  claim 46 , wherein said third party antigen or antigens is selected from the group consisting of third party cells, a cell antigen, a viral antigen, a bacterial antigen, a protein extract, a purified protein, a synthetic peptide presented by autologous antigen presenting cells and a synthetic peptide presented by non-autologous antigen presenting cells.  
   
   
       51 . The method of  claim 50 , wherein said third party cells are allogeneic or xenogeneic with respect to the recipient.  
   
   
       52 . The method of  claim 51 , wherein said third party cells have HLA antigens which are different from HLA antigens of the non-syngeneic donor and which are not cross reactive with HLA antigens of the recipient.  
   
   
       53 . The method of  claim 51 , wherein said third party cells are stimulatory cells selected from the group consisting of cells purified from peripheral blood lymphocytes, cells purified from spleen, cells purified from lymph nodes, cytokine-mobilized PBLs and in vitro expanded antigen-presenting cells (APC).  
   
   
       54 . The method of  claim 46 , wherein affinity purification is effected by an antibody.  
   
   
       55 . The method of  claim 54 , wherein the antibody is selected from the group consisting of anti-CD69, anti-CD25, anti-IL-2 and anti-INFγ.  
   
   
       56 . A cell preparation for preventing or decreasing graft rejection in a human recipient of a transplant, wherein the recipient has been conditioned under lethal or sublethal conditions, wherein the transplant is derived from a non-syngeneic donor wherein the cell preparation comprises non-syngeneic donor derived non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), the cell preparation comprising non-syngeneic donor derived non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), and wherein the cell preparation does not comprise T-lymphocytes capable of developing into anti-recipient CTLs and does not comprise CD4+ cells and/or CD56+ cells; the cell preparation having been generated by a method which comprises: directing peripheral blood lymphocytes derived from the non-syngeneic donor against a third party antigen or antigens, thereby generating third party antigen-treated lymphocytes which comprise anti-third party CTLs; depleting said peripheral blood lymphocytes derived from the non-syngeneic donor of CD4+ cells and/or CD56+ cells; and depleting, by affinity purification, said third party antigen-treated lymphocytes of T-lymphocytes capable of developing into anti-recipient CTLs.  
   
   
       57 . The method of  claim 56 , wherein the non-syngeneic donor is allogeneic with respect to the recipient.  
   
   
       58 . The method of  claim 56 , wherein the non-syngeneic donor is xenogeneic with respect to the recipient.  
   
   
       59 . The cell preparation of  claim 56 , wherein said third party antigen or antigens is selected from the group consisting of third party cells, a cell antigen, a viral antigen, a bacterial antigen, a protein extract, a purified protein, a synthetic peptide presented by autologous antigen presenting cells and a synthetic peptide presented by non-autologous antigen presenting cells.  
   
   
       60 . The cell preparation of  claim 59 , wherein said third party cells are allogeneic or xenogeneic with respect to the recipient.  
   
   
       61 . The cell preparation of  claim 60 , wherein said third party cells have HLA antigens which are different from HLA antigens of the non-syngeneic donor and which are not cross reactive with HLA antigens of the recipient.  
   
   
       62 . The cell preparation of  claim 60 , wherein said third party cells are stimulatory cells selected from the group consisting of cells purified from peripheral blood lymphocytes, cells purified from spleen, cells purified from lymph nodes, cytokine-mobilized PBLs and in vitro expanded antigen-presenting cells (APC).  
   
   
       63 . The cell preparation of  claim 56 , wherein the cell preparation further comprises immature hematopoietic cells derived from the non-syngeneic donor.  
   
   
       64 . The cell preparation of  claim 63 , wherein said immature hematopoietic cells are derived from a source selected from the group consisting of bone marrow, mobilized peripheral blood, fetal liver, yolk sac and cord blood.  
   
   
       65 . The cell preparation of  claim 64 , wherein said cells derived from mobilized peripheral blood are obtained by leukapheresis of peripheral blood of the non-syngeneic donor after stimulation of the non-syngeneic donor with a suitable cytokine.  
   
   
       66 . The cell preparation of  claim 63 , wherein said immature hematopoietic cells are T-cell depleted hematopoietic progenitor cells.  
   
   
       67 . The cell preparation of  claim 63 , wherein said immature hematopoietic cells are CD34+ cells.  
   
   
       68 . The cell preparation of  claim 63 , wherein a cell ratio between said cytotoxic T-lymphocytes and said immature hematopoietic cells including stem cells is at least 1 to 100.  
   
   
       69 . The cell preparation of  claim 63 , wherein said immature hematopoietic cells are stem cells.  
   
   
       70 . The method of  claim 56 , wherein affinity purification is effected by an antibody.  
   
   
       71 . The method of  claim 70 , wherein the antibody is selected from the group consisting of anti-CD69, anti-CD25, anti-IL-2 and anti-INFγ.  
   
   
       72 . A method of producing a cell preparation for preventing or decreasing graft rejection in a human recipient of a transplant, wherein the recipient is conditioned under lethal or sublethal conditions, wherein the transplant is derived from a non-syngeneic donor, wherein the cell preparation comprises non-alloreactive anti-third party cytotoxic T-lymphocytes (CTLs), wherein the cell preparation does not comprise T-lymphocytes capable of developing into anti-recipient CTLs, and wherein the cell preparation does not comprise CD4+ cells and/or CD56+ cells, the method comprising: 
 (i) directing peripheral blood lymphocytes derived from the non-syngeneic donor against a third party antigen or antigens, thereby generating third party antigen-treated lymphocytes which comprise anti-third party CTLs;    (ii) depleting said peripheral blood lymphocytes derived from the non-syngeneic donor of CD4+ cells and/or CD56+ cells; and    (iii) depleting, by affinity purification, said third party antigen-treated lymphocytes of T-lymphocytes capable of developing into anti-recipient CTLs, thereby producing the cell preparation.    
   
   
       73 . The method of  claim 72 , wherein the non-syngeneic donor is allogeneic with respect to the recipient.  
   
   
       74 . The method of  claim 72 , wherein the non-syngeneic donor is xenogeneic with respect to the recipient.  
   
   
       75 . The method of  claim 72 , wherein step (ii) is effected by depleting said peripheral blood lymphocytes derived from the non-syngeneic donor of CD56+ cells or CD4+ cells.  
   
   
       76 . The method of  claim 72 , wherein said third party antigen or antigens is selected from the group consisting of third party cells, a cell antigen, a viral antigen, a bacterial antigen, a protein extract, a purified protein, a synthetic peptide presented by autologous antigen presenting cells and a synthetic peptide presented by non-autologous antigen presenting cells.  
   
   
       77 . The method of  claim 76 , wherein said third party cells are allogeneic or xenogeneic with respect to the recipient.  
   
   
       78 . The method of  claim 77 , wherein said third party cells have HLA antigens which are different from HLA antigens of the non-syngeneic donor and which are not cross reactive with HLA antigens of the recipient.  
   
   
       79 . The method of  claim 77 , wherein said third party cells are stimulatory cells selected from the group consisting of cells purified from peripheral blood lymphocytes, cells purified from spleen, cells purified from lymph nodes, cytokine-mobilized PBLs and in vitro expanded antigen-presenting cells (APC).  
   
   
       80 . The method of  claim 72 , wherein affinity purification is effected by an antibody.  
   
   
       81 . The method of  claim 80 , wherein the antibody is selected from the group consisting of anti-CD69, anti-CD25, anti-IL-2 and anti-INFγ.