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 method of generating antigen-specific T lymphocytes that recognize at least one antigen from two or more different viruses, comprising the step of:
 directly stimulating PBMCs comprising antigen-specific T lymphocytes that recognize at least one epitope from a viral antigen of at least two different viruses with more than one library of peptides, wherein peptides in each library comprise a sequence of amino acids spanning all or part of at least one epitope of at least one antigen of each different virus, thereby expanding antigen-specific T lymphocytes capable of recognizing a cell that expresses part or all of at least one antigen from the different viruses, wherein the antigen-specific T lymphocytes comprise CD4+ T-lymphocytes and CD8+ T-lymphocytes. 
 
     
     
       2. The method of  claim 1 , wherein the viruses are selected from the group consisting of EBV, CMV, Adenovirus, BK, HHV6, RSV, Influenza, Parainfluenza, Bocavirus, Coronavirus, LCMV, Mumps, Measles, Metapneumovirus, Parvovirus B, Rotavirus, West Nile Virus, and a combination thereof. 
     
     
       3. The method of  claim 1 , wherein the stimulating step occurs in a gas permeable cell culture device. 
     
     
       4. The method of  claim 3 , wherein the gas permeable cell culture device comprises a medium that comprises IL-4, IL-7, IL-2, IL-15, IL-12, or a combination thereof. 
     
     
       5. The method of  claim 1 , wherein the T-lymphocytes are administered to an individual. 
     
     
       6. The method of  claim 5 , wherein the individual is an immunocompromised individual. 
     
     
       7. The method of  claim 5 , wherein the individual has lymphoma or leukemia. 
     
     
       8. The method of  claim 5 , wherein the cells are administered by injection. 
     
     
       9. The method of  claim 8 , wherein the injection is intravenous. 
     
     
       10. The method of  claim 5 , wherein the individual has had an allogeneic stem cell transplant. 
     
     
       11. The method of  claim 1 , wherein the stimulating occurs in the presence of IL-4, IL-7, IL-2, IL-15, IL-12, or a combination thereof. 
     
     
       12. The method of  claim 11 , wherein the stimulating occurs in the presence of IL-4, IL-7, or a combination thereof. 
     
     
       13. The method of  claim 5 , wherein at least one epitope is from an adenovirus antigen. 
     
     
       14. The method of  claim 5 , wherein at least one epitope is from the hexon antigen. 
     
     
       15. The method of  claim 5 , wherein at least one epitope is from the penton antigen.