人类疱疹病毒7型表面糖蛋白gH、gL及CD4关系的探讨

发布时间：2018-05-09 18:31

本文选题：HHV-7 + gH　；参考：《南京医科大学》2005年博士论文

【摘要】：人类疱疹病毒7型(Human herpesvirus 7,HHV-7)是1990年首先从健康人外周血单个核细胞中分离出的一种嗜淋巴细胞疱疹病毒。在人群中感染率高,主要通过唾液传播,初次感染多发生在幼儿时期,且病毒在体内可长期持续存在。HHV-7在临床上可能与婴幼儿急疹、玫瑰糠疹及肾脏移植后的巨细胞病毒综合征等有关。 HHV-7与HHV-6同属β-疱疹病毒亚科,两者DNA序列同源性可达到60%,相应氨基酸序列的同源性也达到53.4%,在宿主范围、感染方式、导致的临床症状等方面有一定的相似性。在对HHV-6糖蛋白的研究过程中发现,gH(与gL结合形成复合物)是HHV-6受体CD46的结合蛋白,在HHV-6入侵宿主细胞的过程起着重要的作用。针对其他疱疹病毒的糖蛋白研究也发现gH蛋白(与gL相互结合形成复合物)在病毒与细胞膜的融合以及感染细胞与邻近细胞膜融合过程中起着重要作用。已知CD4分子是HIV感染细胞的重要受体,也是HHV-7感染的主要受体。HIV的gp120能够与CD4结合,介导HIV入侵宿主细胞的过程。HHV-7与CD4结合的病毒蛋白尚未发现。寻找HH-7编码的,与CD4结合的病毒糖蛋白,将为HIV的防治提供新思路。HHV-7糖蛋白gH是否能够与gL结合形成复合物,其能否在病毒入侵中发挥功能,能否与CD4分子相互作用,是本论文的主要研究内容。本研究获得了HHV-7编码gH、gL糖蛋白的基因及CD4的编码基因,应用酵母双杂交技术和免疫共沉淀方法,研究了gH、gL蛋白和CD4的相互作用,并且探讨了gH、gL对HHV-7感染引起CPE的影响。研究取得如下主要结果: 一、HHV-7的培养与鉴定分别接种HHV-7Glasgow标准株于人的脐血单个核细胞(CBMC)和SupT1细胞系。病毒接种后10～15天,镜检出现明显细胞病变效应(CPE)时,以培养物DNA为模板,通过特异性引物扩增,得到493bp的PCR产物。PCR产物测序结果与Genbank中HHV-7RK株、JI株相应序列比对,同源性达到99.9%～100%。
[Abstract]:Human herpesvirus 7 (HHV-7) was first isolated from peripheral blood mononuclear cells (PBMC) of healthy people in 1990. The infection rate is high in the population, mainly transmitted by saliva. The first infection occurs in early childhood, and the virus can persist in the body for a long time. HHV-7 may be clinically associated with acute rash in infants. Pityriasis roseus and cytomegalovirus syndrome after kidney transplantation and so on. HHV-7 and HHV-6 belong to the subfamily of 尾 -herpesvirus. The homology of DNA sequence and amino acid sequence between them can reach 600.The homology of the corresponding amino acid sequence is 53.4. there are certain similarities in host range, infection mode, clinical symptoms and so on. In the study of HHV-6 glycoprotein, it was found that GH (binding to gl) is the binding protein of HHV-6 receptor CD46, which plays an important role in the invasion of host cells by HHV-6. Studies on glycoproteins of other herpes viruses have also found that gh proteins (which bind to gl to form complexes) play an important role in the fusion of viruses with cell membranes and infected cells with adjacent cell membranes. It is known that CD4 molecule is an important receptor of HIV infected cells. It is also the main receptor of HHV-7 infection. HIV gp120 can bind to CD4, and the viral protein that mediates HIV invading host cells. HHV-7 binds to CD4 has not been found. Searching for the viral glycoprotein encoded by HH-7 and binding to CD4 will provide a new idea for the prevention and treatment of HIV. Can the glycoprotein G H of HHV-7 bind to GL to form a complex, can it function in virus invasion, and can it interact with CD4 molecule? Is the main research content of this thesis. In this study, the gene encoding gHG L glycoprotein by HHV-7 and the gene encoding CD4 were obtained. The interaction between gHG L protein and CD4 was studied by yeast two-hybrid technique and immunoprecipitation method, and the effect of gHGL on CPE induced by HHV-7 infection was investigated. The main results of the study are as follows: Culture and Identification of HHV-7 HHV-7Glasgow standard strains were inoculated into human umbilical cord blood mononuclear cells (CBMC) and SupT1 cell lines respectively. When the virulent cytopathic effect was detected by microscope 10 ~ 15 days after inoculation, using the culture DNA as template, the PCR products of 493bp were amplified by specific primers, and the sequencing results of PCR products were compared with the corresponding sequences of HHV-7RK strain JI in Genbank. The homology reached 99.9%.
【学位授予单位】：南京医科大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392

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