手足口病EV71病毒串联表位的免疫性研究

发布时间：2018-10-23 14:08

【摘要】：肠道病毒71型(Enterovirus71，EV71)是小核糖核酸病毒科肠道病毒属，单链的小RNA肠道病毒，其感染性强且致病率高，对中枢神经系统的侵害尤其严重。近年来，EV71引起社会各界的关注是因为发现该病毒是儿童手足口病(HFMD)的主要病原体之一，特别是导致6岁以下儿童重症手足口病（HFMD）的发生。据统计，目前手足口病90%以上的重症病例是由EV71引起，患儿除了有HFMD的典型症状外，还有高达11．1％和0．6％的感染者患有无菌性脑膜炎、脑炎的中枢神经系统疾病，且病程进展迅速，导致近年来死亡率都高于25/1000。EV71的爆发流行严重危害我国儿童的身体健康，给广大民众带来极大的心理恐慌。现阶段国内外尚未研制出针对EV71的疫苗、特异性治疗药物，，对于控制HFMD的发生主要依靠对低幼龄儿童进行宣传教育，建立良好的卫生习惯，因此研究安全，高效，经济的疫苗迫在眉睫。本研究以EV71表位肽疫苗研究为着手点，通过比较29个国内流行株（来自福建，安徽阜阳，江苏，浙江等省份）C4型基因序列，以及28个NCBI基因库中收录的B型EV71异型毒株的基因序列，发现5个表位在不同血清型的病毒株中能保持很高的同源性。以日本脑炎的D蛋白为载体蛋白与上述5个表位以T-T-B和T-B-B的方式串联，一共设计构建了8种表位肽组合，并在原核表达系统中进行了高效表达。以原核表达的重组蛋白为抗原进行的免疫性试验，结果显示，上述8种组合均能在小鼠血清中的产生抗体；其中C，E，F，H四种组合能产生较高水平的抗体，达到本研究的设计目的，期望可为EV71新型疫苗设计提供理论基础，研制出有应用价值的EV71表位肽疫苗。
[Abstract]:Enterovirus 71 (Enterovirus71,EV71) is a small RNA enterovirus belonging to small ribonucleic acid virus family. It is highly infectious and has a high pathogenicity, especially to the central nervous system (CNS). In recent years, EV71 has attracted the attention of all walks of life because it has been found to be one of the major pathogens of HFMD in children, especially (HFMD) in children under 6 years of age. According to statistics, at present, more than 90% of severe cases of hand, foot and mouth disease are caused by EV71. In addition to the typical symptoms of HFMD, 11. 1% and 0. 6% of the infected patients also suffer from aseptic meningitis and encephalitis central nervous system diseases. The rapid progress of the course of disease has led to the death rate higher than the outbreak of 25/1000.EV71 in recent years the outbreak of epidemic serious harm to the health of children in our country and bring great psychological panic to the general public. At present, there has not been developed a vaccine for EV71 and a specific therapeutic drug at home and abroad. In order to control the occurrence of HFMD, it mainly depends on the propaganda and education of young children and the establishment of good hygiene habits. Therefore, the research is safe and efficient. Economic vaccines are imminent. In this study, EV71 epitope peptide vaccine was used as the starting point to compare the C4 gene sequences of 29 prevalent strains (from Fujian, Fuyang, Jiangsu and Zhejiang provinces) in China. The gene sequences of B type EV71 heterotypic strains collected from 28 NCBI gene banks showed that 5 epitopes could maintain high homology in different serotype strains. Eight epitope peptides were designed and expressed in prokaryotic expression system by using Japanese encephalitis D protein as carrier protein in tandem with the above five epitopes by T-B and T-B-B. The results of immunological tests using prokaryotic expressed recombinant protein as antigen showed that all the above eight combinations could produce antibodies in mouse serum, among which four combinations of CfG and FG could produce a higher level of antibodies, so as to achieve the purpose of the design of this study. It is expected to provide a theoretical basis for the design of new EV71 vaccine and to develop EV71 epitope peptide vaccine.
【学位授予单位】：河南师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.1

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