Methods of using il-21 for adoptive immunotherapy and identification of tumor antigens

Abstract

Methods for preparing ex vivo T cell cultures using IL-21 compositions for use in adoptive immunotherapy are described. Addition of IL-21 to cultures of non-terminally differentiated T cells population, either isolated or present in peripheral blood mononuclear cells are exposed to one or more tumor antigens, and in the presence of IL-21 compositions and antigen presenting cells (APCs), the resulting T cell population has an enhanced antigen-specificity, and can be reintroduced into the patient. Methods are also disclosed for identifying tumor antigens by culturing T cell populations exposed to IL-21 compositions and APCs in the presence of tumor material.

1 . A method for identifying a tumor antigen comprising:
 co-culturing tumor material isolated from a subject with peripheral blood mononuclear cells (PBMCs) in the presence of an IL-21 composition and antigen presenting cells (APCs);   isolating a T cell population from the culture;   cloning individual T cells from the T cell population; and   characterizing T cell clones for antigen-specificity.   
     
     
         2 . The method of  claim 1 , wherein the T cell population is a T cell population that is non-terminally differentiated. 
     
     
         3 . The method of  claims 1 , wherein the PBMCs or T cell population is an autologous cell population. 
     
     
         4 . The method of  claim 1 , wherein the tumor material comprises total RNA, lysed tumor cells or apoptotic bodies. 
     
     
         5 . The method of  claim 2 , wherein the isolated T cell population does not include CD4+ cells. 
     
     
         6 . The method of  claim 1 , wherein co-culturing is in the presence of the IL-21 composition and one or more additional cytokines 
     
     
         7 . A method of preparing a naïve CD8+ human T cell population for in use adoptive immunotherapy comprising:
 isolating peripheral blood mononuclear cells (PBMCs) from a biological sample;   identifying PBMCs having a histocompatible phenotype to a subject having a tumor;   isolating a naïve CD8+ human T cell population;   co-culturing tumor material isolated from the subject with the naïve CD8+ human T cell population in the presence of an IL-21 composition and antigen presenting cells (APCs); and   expanding the cells in culture.   
     
     
         8 . The method of  claim 7 , wherein the PBMCs are autologous. 
     
     
         9 . The method of  claim 7 , wherein after the step of co-culturing the tumor material and with the T cell population, the T cells are enriched. 
     
     
         10 . A method of preparing a naïve CD8+ human T cell population for use in adoptive immunotherapy comprising:
 isolating a biological sample containing T cells;   identifying a T cell population having a histocompatible phenotype to a subject having a tumor;   isolating a naïve CD8+ human T cell population;   co-culturing tumor material from the subject with the naïve CD8+ human T cell population in the presence of an IL-21 composition and APCs; and   expanding these cells in culture.   
     
     
         11 . The method of  claim 10 , wherein the T cell population is autologous. 
     
     
         12 . The method of  claim 7 ,  10 , or  16  wherein the tumor material comprises total RNA, lysed tumor cells or apoptotic bodies. 
     
     
         13 . The method of  claim 10 , wherein after the step of co-culturing the tumor material and with the T cell population, the T cells are enriched. 
     
     
         14 . A method for ex vivo expansion of antigen-specific cytotoxic T cells comprising:
 isolating a biological sample containing T cells;   identifying a T cell population having a histocompatible phenotype to a subject;   co-culturing antigenic material from the subject with the T cell population in the presence of an IL-21 composition;   expanding the cells in culture; and   and reintroducing the cells back into the subject.   
     
     
         15 . The method of  claim 14 , wherein after the step of co-culturing the tumor material and with the T cell population, the T cells are enriched. 
     
     
         16 . A method of preparing a non-terminally differentiated CD8+ human T cell population for use in adoptive immunotherapy comprising:
 isolating a biological sample containing T cells;   identifying a T cell population having a histocompatible phenotype to a subject having a tumor;   isolating a non-terminally differentiated CD8+ human T cell population;   co-culturing tumor material from the subject with the non-terminally differentiated CD8+ human T cell population in the presence of an IL-21 composition and APCs; and   expanding these cells in culture.   
     
     
         17 . The method of  claim 16 , wherein the T cell population is autologous.