Abstract: Hepatitis C virus (HCV) is an important cause of acute and chronic hepatitis which is a disorder with a high worldwide prevalence. HCV core protein was considered as immunogenic counterpart of the HCV vaccine and it is an ideal candidate for HCV vaccine. Since cytokines such as IL-6, TNF-α and IFN-Gamma are responsible for the prevention of viral infection, this study aimed to evaluate the effectiveness of HCV core protein as a vaccine. Ten BALB/c mice were immunized with HCV core protein and after 42 days the splenocytes were isolated and the IL-6 and INF-γ secretion were measured using ELISpot technique, at the same time TNF-α was determined by ELISA in the sera. The MTT assay was done to assess the viability of the cultured splenocytes. For evaluating the humoral immune response against the recombinant HCV core protein the DOT Blot test was used. Data was compared using one-way ANOVA test and significant results were considered at p<0.05. In the present study the IL-6, INF-γ and TNF-α levels were dramatically higher in the immunized mice compared to the control group (respectively, 22.9±1.26; 18.53±3.87; 53.96±4.54 and p<0.05). The immunized mice with recombinant HCV core protein showed higher amount of IL-6, INF-γ and TNF-α in the current study. Since the level of IL-6, TNF-α and IFN-γ is high in patients with acute HCV infection, thus a vaccine which could stimulate the secretion of these cytokines in advance may have a preventive role.