Generation of ctl lines with specificity against multiple tumor antigens or multiple viruses

Abstract

The present invention encompasses methods and compositions for the generation and use of cytotoxic T lymphocytes that target multiple viruses or that are specific for multiple tumor antigens. In specific embodiments, the generation methods employ use of certain cytokines to promote proliferation and reduce cell death in an activated T cell population and/or that employ a particular bioreactor having a gas permeable membrane.

What is claimed is: 
     
         1 . A polyclonal population of cytotoxic T-lymphocytes (CTLs) that recognize at least one epitope from each of at least two different tumor antigens, said population comprising CD4+ T-lymphocytes and CD8+ T-lymphocytes. 
     
     
         2 . The population of  claim 1 , wherein the CTLs have been stimulated by dendritic cells (DCs) and/or PBMCs contacted with at least two libraries of peptides that represent the at least two tumor antigens, wherein the peptides in each library overlap in sequence to span at least a part of a tumor antigen. 
     
     
         3 . A composition comprising a population of CTLs that recognize at least one epitope from each of at least two different tumor antigens, said population comprising CD4+ lymphocytes and CD8+ T-lymphocytes. 
     
     
         4 . The composition of  claim 3 , wherein the CTLs have been stimulated one or more times by DCs or PBMCs contacted with at least two libraries of peptides that represent the at least two tumor antigens, wherein the peptides in each library overlap in sequence to span at least a part of a tumor antigen. 
     
     
         5 . The composition of  claim 3 , wherein the CTLs are cultured ex vivo in the presence of IL-7 and IL-15. 
     
     
         6 . The composition of  claim 3 , wherein the CTLs are cultured ex vivo in the presence of IL-7, IL-15, IL-12, and either IL-6 or IL-27. 
     
     
         7 . The composition of  claim 3 , wherein the CTLs are expanded in a gas permeable vessel. 
     
     
         8 . The composition of  claim 4 , where the one or more stimulations has been performed using DCs and where DCs have been used in the first antigen stimulation. 
     
     
         9 . The composition of  claim 4 , wherein the libraries of peptides were chemically synthesized. 
     
     
         10 . The composition of  claim 3 , wherein the population comprises MHC-restricted CTLs. 
     
     
         11 . A method of treating a human patient having a tumor that expresses two or more of PRAME, WT1, Survivin, NY-ESO-1, MAGEA4 and SSX2, comprising administering to the human patient an effective amount of a tumor antigen specific T-cell population that is activated to at least two tumor associated antigens selected from the group of consisting of PRAME, WT1, Survivin, NY-ESO-1, MAGEA4 and SSX2. 
     
     
         12 . The method of  claim 11 , wherein the activation of the population is performed with at least two libraries of peptides that represent the at least two tumor antigens, wherein the peptides in each library overlap in sequence to span at least a part of a tumor antigen. 
     
     
         13 . A method of producing the population of  claim 1 , comprising the steps of stimulating DCs or PBMCs with at least two libraries of peptides that each represent at least one epitope from each of at least two different tumor antigens. 
     
     
         14 . A method of treating a human, comprising administering to the human an effective amount of a T-cell population that is activated to at least two tumor associated antigens selected from the group consisting of PRAME, WT1, Survivin, NY-ESO-1, MAGEA4 and SSX2. 
     
     
         15 . The method of  claim 14 , wherein said human has a cancer that is a solid tumor. 
     
     
         16 . The method of  claim 14 , wherein said human has pancreatic cancer. 
     
     
         17 . The method of  claim 14 , wherein said human has a hematologic cancer.