论文部分内容阅读
目的:构建含有汉坦病毒M基因(编码糖蛋白G2)和部分S基因(编码核蛋白主要抗原区段)以及S基因的多个CTL表位的多种重组腺病毒表达载体,并对这些重组腺病毒的免疫学特性进行研究。方法:构建含目的基因的重组腺病毒载体,并在VeroE6细胞中进行表达,利用纯化后的重组腺病毒免疫BALB/c小鼠,通过ELISA、微量细胞中和试验、T淋巴细胞增殖实验及CTL杀伤实验等方法检测免疫反应。结果:成功表达出可被抗汉滩病毒核蛋白及糖蛋白G2的特异性单克隆抗体(mAb)所识别的融合蛋白;重组的腺病毒可以有效诱导针对汉坦病毒NP及GP的体液免疫应答和细胞免疫应答;同时我们发现含有CTL表位的重组腺病毒比不含CTL表位的重组腺病毒可以诱导小鼠产生更为有效的细胞免疫反应。结论:构建的含CTL多表位的重组腺病毒可以有效的提高嵌合基因刺激细胞免疫应答的能力。
OBJECTIVE: To construct a variety of recombinant adenovirus expression vectors containing multiple CTL epitopes of Hantavirus M gene (encoding glycoprotein G2) and part of S gene (encoding the major antigenic segment of nucleoprotein) and S gene. The immunological characteristics of adenovirus were studied. Methods: The recombinant adenovirus vector containing the gene of interest was constructed and expressed in VeroE6 cells. The purified recombinant adenovirus was used to immunize BALB / c mice. ELISA, microcycle neutralization assay, T lymphocyte proliferation assay and CTL Killing experiments and other methods to detect immune response. Results: The fusion protein recognized by anti-Hantaan virus nucleoprotein and glycoprotein G2-specific monoclonal antibody (mAb) was successfully expressed. The recombinant adenovirus could effectively induce the humoral immune response against Hantavirus NP and GP And cellular immune response. At the same time, we found that the recombinant adenovirus containing CTL epitope could induce more effective cellular immune response in mice than the recombinant adenovirus without CTL epitope. Conclusion: The constructed recombinant adenovirus containing CTL polyepitope can effectively improve the ability of chimeric gene to stimulate cellular immune response.