T cells specific for different latent and lytic viral proteins efficiently control Epstein-Barr virus–transformed B cells

Abstract Background aims Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorders (PTLD) belong to the most dreaded complications of immunosuppression. The efficacy of EBV-specific T-cell transfer for PTLD has been previously shown, yet the optimal choice of EBV-derived antigens inducing polyclonal CD4 + and CD8 + T cells that cover a wide range of human leukocyte antigen types and efficiently control PTLD remains unclear. Methods A pool of 125 T-cell epitopes from seven latent and nine lytic EBV-derived proteins (EBV mix ) and peptide pools of EBNA1, EBNA3c, LMP2a and BZLF1 were used to determine T-cell frequencies and to isolate T cells through the use of the interferon (IFN)-γ cytokine capture system. We further evaluated the phenotype and functionality of the generated T-cell lines in vitro . Results EBV mix induced significantly higher T-cell frequencies and allowed selecting more CD4 + IFN-γ + and CD8 + IFN-γ + cells than single peptide pools. T cells of all specificities expanded similarly in vitro , recognized cognate antigen, and, to a lower extent, EBV-infected cells, exerted moderate cytotoxicity and showed reduced alloreactivity. However, EBV mix -specific cells most efficiently controlled EBV-infected lymphoblastoid cell lines (LCLs). This control was mainly mediated by EBV-specific CD8 + cells with an oligoclonal epitope signature covering both latent and lytic viral proteins. Notably, EBV-specific CD4 + cells unable to control LCLs produced significantly less perforin and granzyme B, probably because of limited LCL epitope presentation. Conclusions EBV mix induces a broader T-cell response, probably because of its coverage of latent and lytic EBV-derived proteins that may be important to control EBV-transformed B cells and might offer an improvement of T-cell therapies.