Methods for off-the-shelf-tumor immunotherapy using allogeneic t-cell precursors

Abstract

The inventive subject matter relates to methods for treating a T-cell deficiency in a subject in need thereof, comprising administering to said subject a T-cell precursor isolated from an allogeneic donor, provided that said allogeneic donor is not MHC-matched to said subject. The inventive methods can be further enhanced by genetic engineering for targeted immunotherapy.

What is claimed is: 
     
         1 . A method for treating a human T-cell deficiency in a subject in need thereof comprising administering to said subject a composition comprising double negative (DN) T cell precursors from an allogeneic donor, wherein said DN T cell precursors consisting essentially of:
 cells having a DN2 CD44 + CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof,   cells having a DN3 (CD44 − CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof, or   cells having a DN2 (CD44 + CD25 + ) phenotype or a phenotype of an equivalent human DN precursor thereof and cells having a DN3 (CD44−CD25+) phenotype or phenotype of an equivalent human DN precursor thereof;   wherein said subject does not receive a separate administration of hematopoietic stem cells;   
       wherein the major histocompatibility complex (MHC) of said allogeneic donor is not matched to the MHC of said subject. 
     
     
         2 . The method of  claim 1 , wherein said DN T-cell precursors are generated by ex vivo culturing of hematopoietic stem cells from the allogeneic donor. 
     
     
         3 . The method of  claim 2 , wherein said DN T-cell precursors are generated by ex vivo culturing said hematopoietic stem cells on a bone marrow derived stromal cell line expressing the Notch ligand Delta-like 1. 
     
     
         4 . The method of  claim 1 , wherein said DN T-cell precursors consist essentially of the cells having the DN2(CD44 + CD25 + ) phenotype or phenotype of the equivalent human DN precursor thereof and the cells having the DN3(CD44 − CD25 + ) phenotype or phenotype of the equivalent DN precursor thereof. 
     
     
         5 . The method of  claim 1 , wherein said DN T-cell precursors are derived from lineage lin − Sca-1 + c-kit hi  or a human equivalent CD34 +  lineage. 
     
     
         6 . A method for treating a human T-cell deficiency in a subject in need thereof, comprising administering to said subject a composition comprising double negative (DN) T-cell precursors from an allogeneic donor, wherein said DN T-cell precursors consist essentially of:
 cells having a DN2 (CD44 + CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof,   cells having a DN3 (CD44 − CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof, or   cells having a DN2 (CD44 + CD25 + ) phenotype or a phenotype of an equivalent human DN precursor thereof and cells having a DN3 (CD44 − CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof,   wherein the major histocompatibility complex (MHC) of said allogeneic donor is not matched to the MHC of said subject, wherein said DN T-cell precursors comprise a nucleic acid sequence which encodes for a chimeric antigen receptor.   
     
     
         7 . The method of  claim 6 , wherein said chimeric antigen receptor is specific for an antigen of a target cancer cell. 
     
     
         8 . The method of  claim 7 , wherein said antigen is CD19. 
     
     
         9 . The method of  claim 8 , wherein said chimeric antigen receptor is 19z1. 
     
     
         10 . The method of  claim 6 , wherein said DN T-cell precursors express one or more proteins selected from the group consisting of costimulatory molecules CD26, CD28, CD40, CD80, CD86, CD134, CD154, and combinations thereof. 
     
     
         11 . The method of  claim 6 , wherein said DN T-cell precursors consist essentially of the cells having the DN2(CD44 + CD25 + ) phenotype or phenotype of the equivalent human DN precursor thereof and the cells having the DN3(CD44 − CD25 + ) phenotype or phenotype of the equivalent DN precursor thereof. 
     
     
         12 . A composition comprising double negative (DN) T-cell precursors from an allogeneic donor, wherein said DN T-cell precursors consist essentially of:
 cells having a DN2 (CD44 + CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof,   cells having a DN3 (CD44 − CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof, or   cells having a DN2 (CD44 + CD25 + ) phenotype or a phenotype of an equivalent human DN precursor thereof and cells having a DN3 (CD44 − CD25 + ) phenotype or phenotype of an equivalent human DN precursor thereof,   wherein said DN T-cell precursors comprise a nucleic acid sequence which encodes for a chimeric antigen receptor.   
     
     
         13 . The composition of  claim 12 , wherein said chimeric antigen receptor is specific for an antigen of a target cancer cell. 
     
     
         14 . The composition of  claim 13 , wherein said antigen is CD19. 
     
     
         15 . The composition of  claim 14 , wherein said chimeric antigen receptor is 19z1. 
     
     
         16 . The composition of  claim 12 , wherein said DN T-cell precursors express one or more proteins selected from the group consisting of costimulatory molecules CD26, CD28, CD40, CD80, CD86, CD134, CD154, and combinations thereof. 
     
     
         17 . The composition of  claim 12 , wherein said DN T-cell precursors are generated by ex vivo culturing of hematopoietic stem cells from the allogeneic donor. 
     
     
         18 . The composition of  claim 17 , wherein said DN T-cell precursors are generated by ex vivo culturing said hematopoietic stem cells on a bone marrow derived stromal cell line expressing the Notch ligand Delta-like 1. 
     
     
         19 . The composition of  claim 12 , wherein said DN T-cell precursors consist essentially of the cells having the DN2(CD44 + CD25 + ) phenotype or phenotype of the equivalent human DN precursor thereof and the cells having the DN3(CD44 − CD25 + ) phenotype or phenotype of the equivalent DN precursor thereof. 
     
     
         20 . The composition of  claim 12 , wherein said DN T-cell precursors are derived from lineage lin − Sca-1 + c-kit hi  or a human equivalent CD34 +  lineage.