HSP70增强汉滩病毒NP和S基因诱导的特异性抗HTNV NP免疫反应的研究

发布时间：2018-04-05 17:17

本文选题：汉滩病毒　切入点：S基因　出处：《第四军医大学》2006年博士论文

【摘要】：HSP70是一个高度保守的蛋白，其在蛋白的合成、折叠、转运以及抗原的加工递呈过程中均具有重要作用。研究表明，HSP70可作为一种分子载体，增强与其结合蛋白的抗原性。汉滩病毒是布尼亚病毒科汉坦病毒(Hantaviruses，HV)的血清型之一，为单股负链RNA病毒，其基因组有大(L)、中(M)、小(S)3个片段组成，分别编码病毒相关的依赖RNA的RNA聚合酶、囊膜糖蛋白(glycoprotein，GP)G1和G2及核衣壳蛋白(nucleocapsidprotein．NP)四种结构蛋白。其中NP具有很强的抗原性和免疫原性，刺激机体产生抗体出现时间早、滴度高、持续时间长，而且也是CTLs(CytotoxicT lymphocytes)识别的主要抗原之一。本实验室以往研究发现，HTNV感染的动物体内存在HSP70-NP复合物。基于NP的汉坦病毒免疫一直受到广泛研究，而将HSP70与汉滩病毒76-118株S基因相结合用于抗HTNV免疫尚未见报道。本研究构建了HSP70与HTNV S融合蛋白疫苗与基因疫苗，以HSP70为疫苗载体(免疫佐剂)，HTNV NP为靶抗原，研究疫苗对机体体液及细胞免疫作用的影响。 1．HSP70-NP融合蛋白原核表达载体的构建、表达与鉴定分别构建了pGEX-4T-1／HSP70-S，pGEX-4T-1／HSP70，pET28a／S三种原核表达载体。用IPTG对含有pGEX-4T-1／HSP70-S，pGEX-4T-1／HSP70
[Abstract]:HSP70 is a highly conserved protein, which plays an important role in protein synthesis, folding, transport and antigen processing and presentation.The results showed that HSP70 could be used as a molecular carrier to enhance the antigenicity of its binding proteins.Hantavirus is one of the serotypes of HantavirusesHVof Bunyaviridae. It is a single-stranded negative-stranded RNA virus. Its genome is composed of three segments, namely, large Laphne, midribosome and Sus, which encode RNA polymerase dependent on RNA, which is associated with the virus.Glycoprotein glycoprotein (GP) G 1 and G 2 and nucleocapsidprotein. NPs.NP has strong antigenicity and immunogenicity, and it is one of the main antigens recognized by CTLs(CytotoxicT lymphocytes.Previous studies in our laboratory have found that HSP70-NP complexes exist in animals infected with HTNV.The immunization of Hantavirus based on NP has been widely studied, but the combination of HSP70 and S gene of Hantaan virus 76-118 strain has not been reported.In this study, HSP70 and HTNV S fusion protein vaccine and gene vaccine were constructed, and HSP70 was used as vaccine vector to study the effect of the vaccine on humoral and cellular immunity.Three prokaryotic expression vectors pGEX-4T-1 / HSP70-SSP70-pGEX-4T-1 / HSP70pET28a / S were constructed by construction, expression and identification of prokaryotic expression vector of 1.HSP70-NP fusion protein.PGEX-4T-1 / HSP70-Sn pGEX-4T-1 / HSP70 using IPTG
【学位授予单位】：第四军医大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R392

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