Epstein-Barr virus (EBV) provides one of the most informative systems for analysing cytotoxic T lymphocyte responses in humans. The viral infection and its persistence are the results of an alternation of lytic and latent phases that are controlled by the immune response. Using a transient COS transfection assay that permits semi-quantitative estimation of CD8 T cell responses against a large number of HLA/viral protein combinations, we analyzed responses to EBV within a large number of polyclonal T cell lines. This allowed a rapid identification of major epitopes and the demonstration that EBV-specificT cells were mainly directed against a restricted set of immunodominant epitopes, primarily generated during the early lytic cycle. Knowledge of the antigen specificity of CDB T cell responses against EBV should help generate cytotoxic T cell lines to this herpesvirus, and more generally to study the molecular basis of immunodominance.