Induction of Hantaan virus-specific immune responses in C57BL/6 mice by immunization with a modified recombinant adenovirus containing the chimeric gene, GcS0.7
Induction of Hantaan virus-specific immune responses in C57BL/6 mice by immunization with a modified recombinant adenovirus containing the chimeric gene, GcS0.7
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通过使用含有嵌合基因 GcS0.7 的改良重组腺病毒进行免疫，在 C57BL/6 小鼠中诱导汉坦病毒特异性免疫反应
DOI：
10.3892/ijmm.2013.1421
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发表时间：
2013-09-01
影响因子：
5.4
通讯作者：
Xu, Zhi-Kai
中科院分区：
文献类型：
--
作者：
Li, Kai;Li, Pu-Yuan;Xu, Zhi-Kai
汉坦病毒糖蛋白Gc是导致体液免疫应答产生的主要成分之一，而核衣壳蛋白（NP）通过诱导抗体依赖性细胞毒性T细胞参与细胞免疫应答。本研究以汉滩病毒（HTNV）为研究对象，构建了一个融合基因GcS0.7，该基因编码含Gc和截短NP的融合蛋白。将嵌合基因与强大的杂合巨细胞病毒（CMV）增强子/鸡β-肌动蛋白（CAG）启动子或土拨鼠肝炎病毒（WHV）转录后调控元件（WPRE）或两者一起克隆到腺病毒载体中。这两个元件都增加了融合蛋白的表达水平。rAd-GcS 0.7-pCAG组融合蛋白表达量最高，是未修饰腺病毒载体的2.3倍。为进一步评价重组腺病毒诱导的体液免疫和细胞免疫效果，检测免疫小鼠的抗体效价、IFN-γ分泌水平和细胞毒性T细胞比例。rAd-GcS 0.7-pCAG组的HTNV特异性体液免疫和细胞免疫应答最强。通过一系列免疫学试验比较了重组腺病毒与灭活疫苗的免疫原性。在细胞免疫应答方面，rAd-GcS 0.7-pCAG组甚至超过了疫苗对照组。CAG杂合启动子不仅提高了融合蛋白的表达水平，而且提高了融合蛋白的免疫原性，为汉坦病毒疫苗的研究提供了一种有希望的策略。
Hantavirus glycoprotein Gc is one of the main components that contribute to the generation of humoral immune responses, while the nucleocapsid protein (NP) is involved in cellular immune responses through the induction of antibody-dependent cytotoxic T cells. In this study, a chimeric gene, GcS0.7, which encodes a fusion protein containing Gc and truncated NP, was constructed as a candidate for Hantaan virus (HTNV) vaccine development. The chimeric gene was cloned into an adenoviral vector in conjunction with the powerful hybrid cytomegalovirus (CMV) enhancer/chicken beta-actin (CAG) promoter or the woodchuck hepatitis virus (WHV) post-transcriptional regulatory element (WPRE), or both. Both elements increased the expression level of the fusion protein. The rAd-GcS0.7-pCAG group demonstrated the highest fusion protein expression level, with a 2.3-fold increase compared with the unmodified adenoviral vector. To further evaluate the humoral and cellular immunity induced by the recombinant adenovirus, the antibody titers, interferon (IFN)-gamma secretion level and cytotoxic T cell ratio were detected in immunized mice. The strongest HTNV-specific humoral and cellular immune responses were detected in the rAd-GcS0.7-pCAG group. The immunogenicity of these recombinant adenoviruses was compared with that of the inactivated vaccine through a series of immunological assays. In terms of the cellular immune responses, the rAd-GcS0.7-pCAG group even exceeded those induced by the vaccine control. The CAG hybrid promoter improved not only the expression level, but also the immunogenicity of the fusion protein, and may thus provide a promising strategy for HTNV vaccine research.