EV71类病毒颗粒的表达和免疫原性的初步评价

中国生物工程杂志

2013, Vol. 33

Issue (1): 8-13

研究报告

EV71类病毒颗粒的表达和免疫原性的初步评价

郭莉莉, 欧霞, 米锴, 孙茂盛, 李鸿钧

云南省重大传染病疫苗工程技术研究中心云南省重大传染病疫苗研发实验室昆明 650118

Expression of Enterovirus 71 VLP in Baculovirus System and Evaluation of Its Immunogenicity

GUO Li-li, OU Xia, MI Kai, SUN Mao-sheng, LI Hong-jun

Yunnan Engineering Center of Vaccine Research Development on Severe Infecticus Diseases Yunnan Keylaboratory of Vaccine Research & Development on Severe Infections Diseases, Kunming 650118, China

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摘要： 目的:运用Bac-to-Bac杆状病毒表达系统构建EV71类病毒颗粒,并对形成的类病毒颗粒的免疫原性进行初步评价。方法:构建重组杆状病毒表达载体Bacmid-P1-3CD,转染昆虫细胞sf9细胞系,获得携带EV71病毒结构基因P1和3CD蛋白酶基因的重组杆状病毒AcMNPV-P1-3CD。采用免疫荧光、Western blot及电镜观察等方法检测重组病毒表达产物及形态学特征。表达产物免疫动物后用ELISA、中和试验的方法对其免疫效果进行初步评价。结果:免疫荧光证明有特异性表达产物,Western blot结果可见大小约为39kDa的VP1特异性条带。电镜结果可观察到大小约为27nm的颗粒。免疫动物的血清ELISA抗体效价为1:776,中和抗体效价为1:588。结论:利用Bac-to-Bac杆状病毒表达系统能共表达EV71的P1和3CD基因,并能形成EV71类病毒颗粒。经初步评价,形成的EV71 类病毒颗粒具有较好的免疫原性。

关键词： 手足口病; EV71; 类病毒颗粒; Bac-to-Bac杆状病毒表达系统; 免疫评价

Abstract: Enterovirus type (EV71) is an etiologic agent responsible for seasonal epidemics of hand-foot-and-mouth disease and causes outbreaks with significant mortality among young children. Expression of enterovirus EV71 Virus-like Particles(VLP) in Bac-to-Bac Baculovirus Expression System and evaluated its immunogenicity. Recombinant AcMNPV-P1-3CD was obtained by transfecting the Bacmid-P1-3CD into the insect cell line of Sf9. The expressed protein was analyzed by IFA, Western blot methods and electron microscope obseruation, The VLPs of EV71 were observed by Electron microscope, while its immunogenicity was measured by ELISA and Microneutralization. The result reveals that EV71 VLP expressed by Recombinant AcMNPV-P1-3CD has strong immunogenicity, which is helpful for the development of the vaccine against EV71 infection.

Key words: HFMD EV71 VLP Bac-to-bac expression system Immunogenicity

收稿日期: 2012-10-25 出版日期: 2013-01-25

ZTFLH:

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通讯作者: 李鸿钧 E-mail: lihj6912@hotmail.com

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引用本文:

郭莉莉, 欧霞, 米锴, 孙茂盛, 李鸿钧. EV71类病毒颗粒的表达和免疫原性的初步评价[J]. 中国生物工程杂志, 2013, 33(1): 8-13.

GUO Li-li, OU Xia, MI Kai, SUN Mao-sheng, LI Hong-jun. Expression of Enterovirus 71 VLP in Baculovirus System and Evaluation of Its Immunogenicity. China Biotechnology, 2013, 33(1): 8-13.

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I1/8

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[1]	薛玲, 刘江宁, 张耀, 张纯, 王祺, 秦川, 刘永东, 苏志国. EV71多表位抗原亲和纯化及类病毒颗粒胞外自组装[J]. 中国生物工程杂志, 2016, 36(7): 34-40.

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