Department of Microbiology, Qom branch, Islamic Azad University, Qom 37185-364, Iran.
Virol Sin. 2013 Jun;28(3):167-73. doi: 10.1007/s12250-013-3291-z. Epub 2013 May 25.
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.
丙型肝炎病毒(HCV)慢性感染是一个全球性的健康问题,人们投入了大量的努力来开发新型疫苗。有效的疫苗需要针对病毒蛋白产生广泛的免疫反应。为了实现这一目标,我们构建了一种表达非结构 3 (NS3)基因的 DNA 疫苗(pcDNA3.1-HCV-NS3),并在 C57BL/6 小鼠中评估了其免疫反应。在这项研究中,使用 HCV 感染患者 1a 型血清,通过巢式逆转录聚合酶链反应(RT-PCR)方法扩增 NS3 基因。将得到的 NS3 基因亚克隆到 pcDNA3.1 真核表达载体中,并通过 Western blot 检测基因表达。将所得的 DNA 疫苗与白细胞介素-12(IL-12)联合作为佐剂共同给予雌性 C57BL/6 小鼠。在最后一次免疫接种后,评估淋巴细胞增殖、细胞毒性和细胞因子水平以测量免疫反应。我们的数据表明,HCV NS3 DNA 疫苗与 IL-12 联合给药可诱导产生显著水平的 IL-4 和干扰素(IFN)-γ(p<0.05)。与对照组相比,接种疫苗的小鼠的细胞毒性和淋巴细胞增殖反应显著增加(p<0.05)。总之,我们的结果表明,HCV NS3 和 IL-12 的联合给药在小鼠模型中显示出强大的免疫原性。