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含E2-gAD融合基因的丙型肝炎病毒(HCV)DNA疫苗的构建与评价

[Construction and evaluation of hepatitis C virus (HCV) DNA vaccine containing E2-gAD fusion gene].

作者信息

Wen Bo, Deng Yao, Tan Wen-Jie, Ying Xiao, Gao Ji-Ming, Ruan Li

机构信息

Key Laboratory of Model Organism Technology & Application, Wenzhou Medical College, Wenzhou 325035, China.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Feb;24(1):53-5.

PMID:20848852
Abstract

OBJECTIVE

To rational design HCV DNA vaccine candidates and evaluate their specific We design to construct two DNA vaccine candidates, one consists of immunity to HCV in mice.

METHODS

We design to construct two DNA vaccine candidates, one consists of E2 (the envelope glycoprotein 2 of HCV) gene only, the second consists of E2-gAD (Globular Domain of Human Adiponectin) fusion gene via overlapping PCR. Confirm the expression of the DNA vaccines by Western blotting, and then vaccinated by injection of DNA vaccines with gene electrotransfer (GET) in BALB/c mice. The immune response was measured by IFN-gamma ELISPOT.

RESULTS

The DNA vaccine candidate consists of E2-gAD could effectively express in vitro , and it could induced a higher anti-HCV T cell response in mice than the one consists of E2 only.

CONCLUSION

The HCV DNA vaccine consists of E2-gAD fusion can increase the immunity of the E, to some extend, and the research paved a way to develop and optimize the novel HCV DNA vaccine.

摘要

目的

合理设计丙型肝炎病毒(HCV)DNA候选疫苗并评估其在小鼠体内对HCV的特异性免疫。

方法

我们设计构建两种DNA候选疫苗,一种仅由E2(HCV包膜糖蛋白2)基因组成,另一种通过重叠PCR由E2-gAD(人脂联素球状结构域)融合基因组成。通过蛋白质印迹法确认DNA疫苗的表达,然后通过基因电转染(GET)将DNA疫苗注射到BALB/c小鼠体内进行接种。通过干扰素-γ酶联免疫斑点法(IFN-γ ELISPOT)检测免疫反应。

结果

由E2-gAD组成的DNA候选疫苗在体外能有效表达,并且与仅由E2组成的疫苗相比,它能在小鼠体内诱导更高的抗HCV T细胞反应。

结论

由E2-gAD融合组成的HCV DNA疫苗在一定程度上可以增强E2的免疫原性,该研究为开发和优化新型HCV DNA疫苗铺平了道路。

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