Viral clearance in hepatitis C virus (HCV) infection has been correlated with strong, multi-specific and sustained T cell responses. The number of functionally active effector T cells determines the outcome of infection. Only a small number of antigen-specific naïve T cells are originally present. Upon infection, they undergo activation, clonal expansion and differentiation to become effector cells. In this study, we determined the ability of dendritic cells (DCs) to prime T cells in vitro to become effector cells upon stimulation with various TLR ligands or IFNalpha. T cell priming and activation was determined by proliferation and production of effector molecules, IFN-gamma and Granzyme B (GrB). HCV Core-specific T cells showed significant increase in proliferation, and the number of HCV Core-specific CD4+ and CD8+ T cells producing IFN-gamma and GrB was higher than control or NS3-specific T cells. These in vitro-primed CD4+ and CD8+ T cells exhibit the phenotype of just-activated and/or armed effector lymphocytes confirming the transition of naïve T cells to effector cells. This is the first study demonstrating the activation of GrB+CD4+ T cells against antigen(s) derived from HCV. Our study suggests a novel role of CD4+ T cells in immunity against HCV.