1. Department of Microbiology, Qom branch, Islamic Azad University, Qom 37185-364, Iran
2. Department of Microbiology, Lahijan Branch, Islamic Azad University, Lahijan 1616, Iran
3. Golestan Research Center of Gastroenterology and Hepatology-GRCGH, Department of Microbiology, Golestan University of Medical Sciences, Gorgan 4917765181, Iran
4. Shefa Neuroscience Research Centre, Tehran 1996835911, Iran
5. Institut für Physiologie I, Westf?lische Wilhelms-Universit?t Münster, Robert-Koch-Strasse Münster 48149, Germany
6. Klinik und Poliklinik für Neurochirurgie, Westf?lische Wilhelms-Universit?t Münster, Münster 48149, Germany
Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral proteins. To achieve this goal, we constructed a DNA vaccine expressing nonstructural 3 (NS3) gene (pcDNA3.1-HCV-NS3) and assessed the immune response in C57BL/6 mice. In this study, the NS3 gene was amplified with a nested-reverse transcriptase-polymerase chain reaction (RT-PCR) method using sera of HCV-infected patients with genotype 1a. The resulting NS3 gene was subcloned into a pcDNA3.1 eukaryotic expression vector, and gene expression was detected by western blot. The resultant DNA vaccine was co-administered with interleukin-12 (IL-12) as an adjuvant to female C57BL/6 mice. After the final immunizations, lymphocyte proliferation, cytotoxicity, and cytokine levels were assessed to measure immune responses. Our data suggest that co-administration of HCV NS3 DNA vaccine with IL-12 induces production of significant levels of both IL-4 and interferon (IFN)-γ (p<0.05). Cytotoxicity and lymphocyte proliferation responses of vaccinated mice were significantly increased compared to control (p<0.05). Collectively, our results demonstrated that co-administration of HCV NS3 and IL-12 displayed strong immunogenicity in a murine model.