EBV Latency II-derived Peptides Induce A Specific CD4+ Cytotoxic T-cell Activity and Not A CD4+ Regulatory T-cell Response

Epstein-Barr virus (EBV) is associated with several malignant diseases that can be distinguished by their patterns of viral latent gene expression. We developed here an original peptidic approach to favor the induction of a specific CD4(+) T-cell response against EBV latency II malignancies (Hodgkin's lymphoma, nasopharyngeal carcinoma, T/NK lymphoma). Previously, we selected 6 peptides derived from EBV nuclear antigen-1, latency membrane proteins (LMP)-1, and LMP-2 highly promiscuous for major histocompatibility complex class II molecules and showed their ability to induce interferon-gamma-secreting CD4(+) T cells. We confirmed here that all peptides used in cocktail are recognized by human CD4(+) memory T cells from healthy donors, inducing a broad T-helper (Th) 1 cytokine secretion interferon-gamma, interleukin-2. Furthermore, we have generated EBV-specific CD4(+) T-cell lines and proved their cytotoxic potential, not only on original models expressing latency II antigens (EBV-transformed T cell or monocyte), but also on lymphoblastoid cell lines expressing latency III antigens (lymphoblastoid cell lines). In addition, granzyme B enzyme-linked immunospot assays suggested that a part of this specific cytotoxic activity could be linked to the granule lytic pathway. Very importantly, we have showed that neither phenotypical changes nor functional activities of CD4(+) CD25(+) CD127(low)-regulatory T cells were observed in response to EBV peptides, avoiding any risk of aggravation of the preexisting immunosuppressive environment reported in EBV+-associated malignancies. In conclusion, our promiscuous peptide cocktail could be used safely in immunotherapeutic approaches against EBV latency II malignancies, mainly to prevent relapse in high-risk patients further to classic treatments.