轮状病毒减毒活疫苗交叉免疫反应的研究

发布时间：2018-03-22 13:50

本文选题：轮状病毒　切入点：交叉保护　出处：《中国药品生物制品检定所》2011年硕士论文　论文类型：学位论文

【摘要】：轮状病毒(Rotavirus, RV)是引起婴幼儿重症腹泻最重要的病原,也是导致发展中国家婴幼儿死亡的主要原因之一。轮状病毒的治疗目前尚无特效药,临床上主要采用补液、维持电解质平衡及对症治疗等方法,只能挽救生命,不能控制病毒的传播。因此开发轮状疫苗成为当今WHO疫苗发展计划中的首要任务。目前国内外批准上市的疫苗有三种:人减毒活疫苗(Rotarix)、人-牛五价重配苗(RotaTeq)和羊减毒活疫苗(LLR)[2-4]。我国羊三价减毒活疫苗正在进行Ⅰ期临床,人-牛六价重配苗也处于研发阶段[5]。由于主要是A组轮状病毒引起儿童重症腹泻,所以这些疫苗的生产用病毒株均为A组。随着疫苗的研制与上市,对于疫苗来说究竟是单价还是多价能起到最好的保护效果?单价的是哪型好?多价的需要多少价才可以最大效果保护好人类?哪种型别的毒株能最大范围的与其他毒株进行交叉免疫保护?研究发现动物与人类毒株之间有在自然界发生重组的可能性。由于目前说法不一,并且缺乏基础研究,对疫苗的评价也造成困难,所以我们开展这个课题,以期解决目前的瓶颈问题。本课题采用已上市的两种种轮状病毒疫苗—葛兰素史克公司的人减毒活疫苗(Rotarix)和兰州生物制品研究所的羊减毒活疫苗(LLR)及轮状病毒Wa株、DS-1株,SA11株及ST-3株作为实验病毒株,进行了轮状病毒免疫交叉反应的研究。本研究就三个方面进行探讨,第一部分为轮状病毒减毒交叉免疫反应的研究,以期找到LLR和Rotarix免疫后抗体能最大范围保护哪种轮状病毒的感染。这些为以后轮状病毒疫苗的研发与质控打下基础;第二部分为从噬菌体展示随机十二肽库中筛选轮状病毒特异性结合肽的实验,挑选出于用轮状病毒免疫后血清结合力强的克隆,将获得的噬菌体克隆进行DNA测序,根据噬菌体克隆基因中所插入的外源DNA序列推导出呈现的外源多肽氨基酸序列,应用DNAStar软件及其它生物信息网络软件如BLAST软件将多肽序列比较分析,进行蛋白结构数据库和文献数据库检索;第三部分为轮状病毒结构蛋白VP5、VP6、VP7、VP8的原核表达与纯化,以期获得轮状病毒融合蛋白,并对其免疫原性进行研究。
[Abstract]:Rotavirus (Rotavirus, RV) is the cause of severe diarrhea in infants and young children the most important pathogen, is one of the main causes of infant death in developing countries. The treatment of rotavirus is currently no cure, the main clinical use of rehydration, electrolyte balance and symptomatic treatment methods can save lives, communication can not control the virus. Therefore the development of rotavirus vaccine has become a primary task in the WHO vaccine development plans.
There are three kinds of domestic licensed vaccines, live attenuated vaccine (Rotarix), human bovine reassortant vaccine quinquevalent (RotaTeq) and sheep live attenuated vaccine (LLR) [2-4]. China sheep trivalent live attenuated vaccine ongoing phase I clinical, human bovine reassortant vaccine in six price the development stage is mainly due to [5. Group A rotavirus cause severe diarrhea, so these vaccine virus strains were produced by A group.
With the development and market of the vaccine, a vaccine for what is the price or multivalent can play a protective effect of the best price? What type? How can the price of polyvalent maximum effect to protect human beings? What types of strains can cross immune protection and the maximum range of other strains in research found? The possibility of recombination occurs in nature between animal and human strains. Due to the inconsistency, and lack of basic research, evaluation of the vaccine is difficult, so we carry out this task, in order to solve the bottleneck problem at present. This paper has listed two kinds of rotavirus vaccine, live attenuated human GlaxoSmithKline Co the vaccine (Rotarix) and the Lanzhou Institute of biological products of sheep live attenuated vaccine (LLR) and rotavirus Wa strain, DS-1 strain, SA11 strain and ST-3 strain as the experimental strains of rotavirus disease Study on the cross reaction of toxic immune system.
This study is to explore three aspects, the first part is the study of rotavirus attenuated cross immune reaction, in order to find the LLR and Rotarix antibody to the largest extent to protect what kind of rotavirus infection. The development and quality control after rotavirus vaccine foundation; the second part is to demonstrate the experiment screening of rotavirus specific binding peptides from phage random twelve peptide library, selected for cloning with rotavirus serum adhesion, the phage clones obtained by DNA sequencing, based on the insertion of exogenous DNA gene in phage clone sequence deduced amino acid sequence of exogenous peptides presented, using DNAStar software and other biological information network software such as BLAST software to comparative analysis of the peptide sequence, protein structure database and document database retrieval; the third part is the rotavirus structural protein VP5 The prokaryotic expression and purification of VP6, VP7 and VP8 were expressed in order to obtain rotavirus fusion protein and to study the immunogenicity of rotavirus.

【学位授予单位】：中国药品生物制品检定所
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.1

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