A组轮状病毒VP5<sup>*</sup>和VP8<sup>*</sup>的表达和免疫学性质研究

中国生物工程杂志

2010, Vol. 30

Issue (02): 32-38

研究报告

A组轮状病毒VP5^*和VP8^*的表达和免疫学性质研究

张艳,文喻玲,魏海涛,李传印,范耀春,陈元鼎^*

中国医学科学院&北京协和医学院医学生物学研究所昆明 650118

Expression of VP5^*and VP8^* of Goup A Rotavirus and Immunological Characteristics

全文: PDF(647 KB) HTML

摘要：

轮状病毒是引起婴幼儿腹泻的主要病原，VP4是RV重要的抗原蛋白,在早期病毒与细胞黏附过程中发挥重要作用，包括受体结合和细胞渗透。在细胞黏附过程中，VP4易被切割成VP5*和VP8*两个片段并以此增强病毒感染性。为了深入研究VP5*和VP8*的免疫学性质，进一步评价其应用前景，本研究从TB-Chen株RV基因组中编码VP4蛋白基因上克隆了VP5*和VP8*开放读码框核苷酸序列，构建了表达质粒，在原核大肠杆菌系统中重组表达了VP5*和VP8*蛋白，进一步分析了它们的免疫学性质。结果显示，VP5*和VP8*可在E.coli中高效表达，重组蛋白VP5* (rVP5*)和VP8* (rVP8*)可诱导免疫豚鼠产生特异性血清抗体，这些抗体可特异性识别自身蛋白（rVP5*或rVP8*），可识别来自的TB-Chen株重组VP4蛋白，并可识别SA11和Wa感染的MA104细胞中合成的病毒VP4蛋白。这些结果表明，rVP5*和rVP8*蛋白具有较高的免疫原性，抗rVP5*和抗rVP8*的抗体具有高度特异性和交叉反应性。

关键词： rotavirus; VP5^*; VP8^*; Recombinant expression; Immunological characteristics

Abstract:

Infections of rotaviruses are the primary cause of severe infantile gastroenteritis worldwide. VP4 is one of the major antigenic protein of the virus. VP4 has an essential role during the early interactions of the virus with the cell surface, including receptor binding and cell penetration. Cleavage of the VP4 protein prior to cell attachment by trypsin-like enzymes into two peptide fragments VP5* and VP8* that remain associated with the virus particle enhances its infectivity. In order to further investigate properties of this two proteins, open reading frame (ORF) sequences of VP5* and VP8* proteins derived from the VP4 of rotavirus strain TB-Chen were cloned and expressed and their immunological characteristics were analyzed. The results showed that VP5* and VP8* proteins could be highly expressed in E. coli cells; the expressed recombinant VP5* (rVP5*) and VP8* (rVP8*) could elicit specific antibodies in guinea pigs, these antibodies could recognize the recombinant VP4 derived from TB-Chen strain and the VP4 protein synthesized in Wa infected or SA11 infected MA104 cells. The results indicated that rVP5* and rVP8* have good immunogenicity and anti-rVP5* and anti-rVP8* antibodies have high specificity and cross reactivity.

Key words: 轮状病毒 VP5^* VP8^* 重组表达抗原性

收稿日期: 2009-09-14 出版日期: 2010-02-26

通讯作者: 陈元鼎 E-mail: chenyd@imbcams.com.cn

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	张艳
	文喻玲
	魏海涛
	李传印
	范耀春
	陈元鼎

引用本文:

张艳文喻玲魏海涛李传印范耀春陈元鼎. A组轮状病毒VP5^*和VP8^*的表达和免疫学性质研究[J]. 中国生物工程杂志, 2010, 30(02): 32-38.

ZHANG Yan, WEN Yu-Ling, WEI Hai-Chao, LI Chuan-Yi, FAN Yao-Chun, CHEN Yuan-Ding. Expression of VP5^*and VP8^* of Goup A Rotavirus and Immunological Characteristics. China Biotechnology, 2010, 30(02): 32-38.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I02/32

［1］ de Zoysa I, Feachem R G. Interventions for the control of diarrhoeal diseases among young children: rotavirus and cholera immunization. Bull WHO, 1985, 63(3):569583.
［2］ Glass R I, Lew J F, Gangarosa R E, et al. Estimates of morbidity and mortality rates for diarrheal diseases in American children. J Pediatr, 1991, 118(4):2733.
［3］ Glass R I, Parashar U D, Bresee J S, et al. Rotavirus vaccines: current prospects and future challenges. Lancet, 2006, 368(9532):323332.
［4］ Parashar U D, Gibson C J, Bresse J S, et al. Rotavirus and severe childhood diarrhea. Emerg Infect Dis, 2006, 12(2):304306.
［5］ Shaw A L, Rothnagel R, Chen D, et al. Threedimensional visualization of the rotavirus hemagglutinin structure. Cell, 1993, 74(4):693701.
［6］ Prasad B V, Chiu W. Structure of rotavirus. Curr Top Microbiol Immunol, 1994, 185:929.
［7］ Yeager M, Berriman J A, Baker T S, et al. Threedimensional structure of the rotavirus haemagglutinin VP4 by cryoelectron microscopy and difference map analysis. EMBO J, 1994, 13(5):10111018.
［8］ Chen Y, Wen Y, Liu X, et al. Full genomic analysis of human rotavirus strain TBChen isolated in China. Virology, 2008, 375(2):361373.
［9］陈元鼎，范耀春，李传印.轮状病毒分类与命名.国际病毒学杂志，2009，16（4）：115120. Chen Y D, Fan Y C, Li C Y. Internatl J Virol, 2009, 16:(4):115120.
［10］文喻玲，李传印，范耀春，等.TBChen株轮状病毒NSP5/NSP6及基因型研究.病毒学报,2009,25(5): 349354. Wen Y L, Li C Y, Fan Y C, et al. Chin J Virology, 2009, 25(5):349354.
［11］ Zhao Q H, Wen Y L, Yu Y, et al. Expression and immunoreactivity of a human group A rotavirus Vp4. Virologica Sinica, 2007, 22(4):287293.
［12］ Arias C F, Guerrero C A, Mendes E, et al. Early events of rotavirus infection: the search for the receptor(s). Novartis Found Symp, 2001, 238:47~60
［13］ Déctor M A, Romero P, López S, et al. Rotavirus gene silencing by small interfering RNAs. EMBO Rep, 2002, 3(12):11751180.
［14］ Clark S M, Roth J R, Clark M L, et al. Trypsin enhancement of rotavirus infectivity: mechanism of enhancement. J Virol, 1981, 39(3):816822.
［15］ Chwetzoff S, Trugnan G. Rotavirus assembly an alternative model that utilizes a typical trafficking pathway. Curr Top Microbiol Immunol, 2006, 309:245261.
［16］ Delmas O, DurandSchneider A M, Cohen J, et al. Spike protein VP4 assembly with maturing rotavirus requires a postendoplasmic reticulum event in polarized caco2 cells. J Virol, 2002, 78(20):1098710994.

No related articles found!

Viewed

Full text

Abstract

Cited

Shared

Discussed