Pepmixes to generate multiviral ctls with broad specificity

Abstract

The present invention concerns methods of generating CTLs that are able to target at least one antigen from two or more viruses. The method includes exposing mixtures of peptides for different antigens to the same plurality of PBMCs and, at least in certain aspects, expanding the cells in the presence of IL4 and IL7.

1 . A method of generating cytotoxic T-lymphocytes (CTLs) that target at least one antigen from two or more viruses, comprising the steps of:
 contacting a plurality of peripheral blood mononuclear cells with at least two libraries of peptides, said libraries of peptides each comprising peptides that correspond to a particular viral antigen; and   expanding the plurality of cells in the presence of one or more cytokines.   
     
     
         2 . The method of  claim 1 , wherein said method occurs in the absence of exposing the libraries to isolated peptide-pulsed dendritic cells prior to expanding the CTLs. 
     
     
         3 . The method of  claim 1 , wherein the one or more cytokines are selected from the group consisting of IL4, IL7 and a combination thereof. 
     
     
         4 . The method of  claim 1 , wherein the peptides are further defined as peptides that overlap in sequence to span part or all of a viral antigen. 
     
     
         5 . The method of  claim 3 , wherein the peptides overlap by at least three amino acids. 
     
     
         6 . The method of  claim 3 , wherein the peptides are at least seven amino acids in length. 
     
     
         7 . The method of  claim 1 , wherein the viruses are selected from the group consisting of EBV, CMV, Adenovirus, BK virus, HHV6, RSV, Influenza, Parainfluenza, Bocavirus, Coronavirus, LCMV, Mumps, Measles, Metapneumovirus, Parvovirus B, Rotavirus, West Nile Virus, JC, HHV7, and a combination thereof. 
     
     
         8 . The method of  claim 1 , wherein the virus is EBV and the antigen is selected from the group consisting of EBNA1, LMP2, and BZLF1. 
     
     
         9 . The method of  claim 1 , wherein the virus is CMV and the antigen is selected from the group consisting of IE1 and pp65. 
     
     
         10 . The method of  claim 1 , wherein the virus is Adv and the antigen is selected from the group consisting of Hexon and penton. 
     
     
         11 . The method of  claim 1 , wherein the virus is BK virus and the antigen is selected from the group consisting of LT and VP-1. 
     
     
         12 . The method of  claim 1 , wherein the virus is HHV6 and the antigen is selected from the group consisting of U14, U11, U71, U54, and U90. 
     
     
         13 . The method of  claim 1 , wherein the virus is RSV and the antigen is selected from the group consisting of N and F. 
     
     
         14 . The method of  claim 1 , wherein the virus is Influenza and the antigen is selected from the group consisting of MP1 and NP1. 
     
     
         15 . The method of  claim 1 , wherein the CTLs are administered to an individual. 
     
     
         16 . The method of  claim 1 , wherein the CTLs are administered to an immunocompromised individual. 
     
     
         17 . The method of  claim 15 , wherein the individual has had allogeneic stem cell transplant. 
     
     
         18 . The method of  claim 14 , wherein the cells are administered by injection. 
     
     
         19 . The method of  claim 17 , wherein the injection is intravenous. 
     
     
         20 . The method of  claim 1 , wherein the CTLs are further defined as polyclonal CD4+ and CD8+ CTLs. 
     
     
         21 . The method of  claim 14 , wherein the PBMCs are allogeneic to the individual. 
     
     
         22 . The method of  claim 14 , wherein the PBMCs are autologous to the individual. 
     
     
         23 . The method of  claim 1 , further comprising the step of exposing the CTLs to one or more compositions that stimulate cell division.