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在C57BL/6小鼠中对表达丙型肝炎病毒非结构蛋白2基因的DNA疫苗进行免疫原性评估。

Immunogenicity evaluation of a DNA vaccine expressing the hepatitis C virus non-structural protein 2 gene in C57BL/6 mice.

作者信息

Gorzin Zahra, Gorzin Ali Akbar, Tabarraei Alijan, Behnampour Naser, Irani Shiva, Ghaemi Amir

机构信息

Dept. of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Shiraz HIV/AIDS Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

Iran Biomed J. 2014;18(1):1-7. doi: 10.6091/ibj.1231.2013.

Abstract

BACKGROUNDS

Most of the hepatitis C virus (HCV) infections elicit poor immune responses and 75% to 85% of cases become chronic; therefore, the development of an effective vaccine against HCV is of paramount importance. In this study, we aimed to evaluate co-administration of HCV non-Structural Protein 2 and IL-12 DNA vaccines in C57BL/6 mice.

METHODS

A plasmid encoding full-length HCV NS2 protein (non-structural protein 2) was generated and used to vaccinate mice. Negative control (an empty expression vector) was also employed to evaluate the background response. To investigate immune responses against vaccine, C57BL/6 mice received three doses of the vaccine with a two-week interval. Cellular immunity was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay for lymphocyte proliferation, lactate dehydrogenase release for cytotoxic T lymphocyte (CTL) activity and cytokine assay.

RESULTS

The findings demonstrated that immunization of mice with plasmid expressing HCV NS2 induced CTL response, interferon gamma production, and lymphocyte proliferation compared to negative control. The results also demonstrated that co-administration of IL-12 with the HCV NS2 plasmid induced significantly better immune response in C57BL/6 mice.

CONCLUSION

DNA vaccine encoding HCV NS2 is an effective candidate that can trigger CTL-based immune response against HCV. In addition, the results suggested that combining the DNA vaccine approach with immune stimulatory cytokines may significantly enhance antigen-specific immune responses.

摘要

背景

大多数丙型肝炎病毒(HCV)感染引发的免疫反应较差,75%至85%的病例会发展为慢性感染;因此,开发一种有效的抗HCV疫苗至关重要。在本研究中,我们旨在评估在C57BL/6小鼠中联合使用HCV非结构蛋白2和IL-12 DNA疫苗。

方法

构建了一种编码全长HCV NS2蛋白(非结构蛋白2)的质粒,并用于给小鼠接种疫苗。还使用阴性对照(空表达载体)来评估背景反应。为了研究针对疫苗的免疫反应,C57BL/6小鼠每隔两周接受三剂疫苗。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法评估淋巴细胞增殖的细胞免疫、通过乳酸脱氢酶释放评估细胞毒性T淋巴细胞(CTL)活性以及进行细胞因子检测。

结果

研究结果表明,与阴性对照相比,用表达HCV NS2的质粒免疫小鼠可诱导CTL反应、干扰素γ产生和淋巴细胞增殖。结果还表明,在C57BL/6小鼠中,IL-12与HCV NS2质粒联合使用可诱导明显更好的免疫反应。

结论

编码HCV NS2的DNA疫苗是一种有效的候选疫苗,可引发针对HCV的基于CTL的免疫反应。此外,结果表明将DNA疫苗方法与免疫刺激细胞因子相结合可能会显著增强抗原特异性免疫反应。

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