Study on molecular mechanisms of herpesvirus entry into host cells

1. 通过细胞-细胞融合实验和VSV假分型实验，证明了单纯疱疹病毒1型（HSV-1）的gB、gH、gL、gD是病毒进入所必需的。在相同的条件下，任何由巨细胞病毒（CMV）和水痘-带状疱疹病毒（VZV）编码的糖蛋白组合都没有产生阳性结果。然而，CMV或VZV糖蛋白中的任何一种都能抑制HSV-1糖蛋白介导的与假型vsv - 2的融合或感染。为了分析糖蛋白相关中和作用的表集，建立了一种新的VSV假型与MLV env蛋白和靶糖蛋白结合的检测方法。利用VZV和CMV.4报告细胞系，对肝素样分子的附着抑制和激酶抑制剂对早期感染的抑制进行了评价。建立了一种基于过滤器的实时PCR检测cmv阳性临床材料的方法。用该方法鉴定的CMV DNA标本进行糖蛋白基因分型。gH、gO和gN基因型之间存在连锁，而gB基因型与其他糖蛋白基因型之间不存在连锁。一种假设是，中和对gB的压力与缺乏连锁有关，另一种假设是，gH、gO和gN的特定组合是它们在病毒进入中发挥作用所必需的。描述了CMV感染激活干扰素刺激基因54K （ISG54K）启动子的启动子区域。激活依赖于CMV IE基因的表达。

1. Using a cell-cell fusion assay and a VSV pseudotyping assay, it was demonstrated that herpes simplex virus type 1 (HSV-1) gB, gH, gL, gD were necessary for virus entry. Under the same condition, any combinations of glycoproteins encoded by cytomegalovirus (CMV) and varicella-zoster virus (VZV) did not yield positive results. However, any of CMV or VZV glycoproteins inhibited the HSV-1 glycoprotein mediated fusion or infection with pseudotyped VSV.2. To analyze epitomes on glycoproteins associated neutralization, a new assay using VSV pseudotyped with MLV env protein and the target glycoprotein.3. Inhibition of attachment with heparin-like molecules and inhibition of immediate early phase of infection with kinase inhibitors were evaluated by using reporter cell lines for VZV and CMV.4. A filter-based real-time PCR assay was developed for identification of CMV-positive clinical materials. CMV DNA specimens identified by the assay were used for genotyping of glycoprotein genes. There was a linkage among gH, gO, and gN genotypes, although there was no linkage of gB genotypes with those of other glycoproteins. One hypothesis is that pressure on gB by neutralization relates to the lack of linkage and alternative hypothesis is that particular combination of gH, gO, and gN are required for their functions in virus entry.5. Promoter region for activation of interferon stimulating gene 54K (ISG54K) promoter by CMV infection was delineated. The activation was dependent on CMV IE gene expression.