Antigen-specific T cells are likely to provide a critical defense against hepatitis C virus (HCV) infection. However, their detection in blood is uncommon except in persons who undergo spontaneous recovery after acute HCV infection. We postulated that virological responses after antiviral interferon-alpha therapy may be associated with enhanced cytolytic T cell immunity. Peripheral blood memory CTL responses were quantified using short term limiting dilution culture, with cytolytic function detected by standard chromium release assay. In this cross-sectional study, 5 of 11 interferon-alpha or interferon-alpha plus ribavirin-treated subjects exhibited cytolytic T cell responses after therapy completion; 4 of these 5 subjects were HCV RNA negative at the time of assay. In contrast, only 1 of 9 untreated chronically viremic subjects had detectable HCV-specific cytolytic T cell responses. Although the requisite factors necessary to achieve sustained virologic response after therapy remain largely undefined, the findings presented here suggest that antiviral therapy-induced virological clearance may be associated with the induction, expansion, and/or recirculation of HCV antigen-specific cytolytic T cells, and may play a role in the maintenance of a nonviremic state.