重组黄热病毒E蛋白结构域Ⅲ作为亚单位疫苗的抗原性和免疫原性  

Immunogenicity and Antigenicity of Recombinant Yellow Fever Virus Envelope Domain Ⅲ as a Subunit Vaccine

作　　者：杨鹏[1,2] 黄莺[1] 刘珊[1] 孙志伟[1] 俞炜源[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]解放军第210医院403临床部检验科,辽宁大连116021

出　　处：《生物技术通讯》2009年第5期635-638,共4页Letters in Biotechnology

摘　　要：目的:表达纯化黄热病毒(YFV)囊膜蛋白(E蛋白)结构域Ⅲ,研究其作为亚单位疫苗预防YFV、日本脑炎病毒(JEV)感染的可能。方法:扩增YFVE蛋白结构域Ⅲ(YFDⅢ)的cDNA片段333bp,将其连接到原核表达载体pET-32a(+)中,构建原核表达载体pET-YFDⅢ,转化感受态大肠杆菌Rosetta(DE3),IPTG诱导表达重组YFDⅢ;用纯化的YFDⅢ免疫新西兰兔和BALB/c鼠,检测相关抗体滴度。结果:在大肠杆菌中可溶性表达了YFDⅢ融合蛋白,表达量约占菌体蛋白的50%;Western印迹及ELISA分析表明,纯化的YFDⅢ具有良好的抗原性和免疫原性;利用纯化的YFDⅢ免疫新西兰兔,获得了高达1∶4×105滴度的抗YFV抗体和1∶2×104滴度的抗JEV抗体;利用纯化的YFDⅢ免疫BALB/c鼠,获得了1∶7×104滴度的抗YFV抗体和1∶2×103滴度的抗JEV抗体。结论:重组YFDⅢ有较好的免疫原性,具有开发成亚单位疫苗的潜能。Objective： To express and purify the envelope domain Ⅲ of yellow fever virus（YFV）（YFDm）, research on the possibility of developing the YFDⅢ as a subunit vaccine against the YFV and Japanese encephalitis virus （JEV）. Methods： The cDNA of the YFDm of the attenuated strain YF-17D was cloned to the pET-32a （＋） expression vector and expressed as a thioredoxin fusion protein in Escherichia coli Rosetta （DE3）. Rabbits and mice were immunized with the putified YFDⅢ, and the relative antibodies were analysed. Results： The recombinant protein was unique in forming a large fraction of the soluble recombinant protein in E.coli. The antigenicity and immunogenicity of the YFDⅢ was tested by Western blot and ELISA. Rabbits immunized with the purified YFDm generated 1：4±10^5 anti-YFV antibody titers and 1：2±10^4 anti-JEV antibody titers. Mice immunized with the purified YFDⅢ generated1 ：7±10^4 anti-YFV antibody titers and 1：2±10^3 anti-JEV antibody titers. Conclusion： The recombinant YFDⅢ delivered in mice and rabbits can generate high antibody titers against YFV and JEV, can serve as a potential vaccine to prevent YFV and JEV infection.

关 键 词：黄热病毒 E蛋白 结构域Ⅲ 免疫原性 抗原性 亚单位疫苗 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]