脂筏在人类疱疹病毒6型装配中的作用及对病毒感染的影响

发布时间：2018-02-06 06:30

本文关键词： 人类疱疹病毒6型脂筏胆固醇感染　出处：《吉林大学》2006年博士论文　论文类型：学位论文

【摘要】：为了探讨脂筏(lipid raft)在人类疱疹病毒6型(HHV-6)的装配与在病毒感染中的作用,用HHV-6 GS株感染HSB2细胞,用非离子去污剂Triton X-100提取脂筏成分,利用Western Blot分析HHV-6包膜糖蛋白与脂筏的相关性。并用免疫荧光双标记的方法,从分子共定位的角度研究HHV-6糖蛋白B(gB)与GPI锚固蛋白CD59分子以及神经节苷脂GM1分子之间的表达与分布关系。结果显示,HHV-6包膜糖蛋白B、H、L、Q1和Q2(gB、gH、gL、gQ1和gQ2)分布在脂筏部位。激光共聚焦显微镜可观察到CD59分子及GM1均与HHV-6包膜糖蛋白B有着相同的分布。另外,在纯化的HHV-6颗粒中检测到了脂筏的指示剂GM1分子。为了探讨构成病毒包膜的脂筏成分在病毒感染中的作用,用甲基-β-环糊精(MβCD)从HHV-6颗粒中去除脂筏的主要组成成分胆固醇,结果显示,病毒的感染呈剂量依赖性降低,而补充外源性胆固醇后可恢复其感染率。用胆固醇提取剂MβCD处理后,HHV-6病毒颗粒虽然能够与靶细胞结合但不能进入到细胞内部。本研究的结果揭示了脂筏提供HHV-6装配的平台,且病毒包膜中的脂筏成分在病毒的感染过程中起着重要的作用。
[Abstract]:In order to investigate the role of lipid raft in the assembly of human herpesvirus 6 (HHV-6) and its role in viral infection, HSB2 cells were infected with HHV-6 GS strain. The components of lipid raft were extracted by non-ionic detergent Triton X-100. The correlation between HHV-6 envelope glycoprotein and lipid raft was analyzed by Western Blot, and immunofluorescence double labeling method was used. The expression and distribution of HHV-6 glycoprotein (HHV-6), GPI anchored protein (CD59) and ganglioside (GM1) molecules were studied from the point of molecular co-localization. The HHV-6 envelope glycoprotein (HHV-6) was found to be Q1 and Q2GBG / gH1, respectively. GQ1 and gQ2) were distributed in lipid rafts. The same distribution of CD59 and GM1 was observed by confocal laser microscopy with HHV-6 coated glycoprotein B. The indicator GM1 molecule of lipid raft was detected in purified HHV-6 particles. In order to investigate the role of lipid raft components that form viral envelope in viral infection, cholesterol was removed from HHV-6 granules by methyl 尾 -cyclodextrin M 尾 CD-CD2. The results showed that cholesterol was the main component of lipid raft. The infection rate of the virus was decreased in a dose-dependent manner, and the infection rate could be recovered after supplementation of exogenous cholesterol, which was treated with cholesterol extractant M 尾 CD. Although HHV-6 virus particles can bind to target cells, they cannot enter the cells. The results of this study revealed that lipid rafts provide a platform for HHV-6 assembly, and the lipid rafts in viral envelope play an important role in the process of virus infection.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R373

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