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Studies on glycoproteins of varicella-zoster virus and their biological functions.

水痘带状疱疹病毒糖蛋白及其生物学功能的研究。

基本信息

批准号：
62480160
负责人：
YAMANISHI Koichi
金额：
$ 3.71万
依托单位：
Research Institute for Microbial Diseases,Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

Varicella-zoster virus (VZV) contains four major glycoproteins designated as gpI,gpII,gpIII and gpIV. Firstly, the presence and role of fatty acids in these glycoproteins were examined. The incorporation of fatty acid into the glycoproteins that span the membranes of VZV was studied using infected cells labeled with [^3H]-palmitic acid. An extract of the cells was treated with monoclonal antibodies to VZV glycoproteins and the precipitate was analyzed by sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The electrophoretic pattern showed that gpI was heavily labeled, and gpII and IV were lightly labeled. Cerulenin, an antibiotic that inhibits de novo fatty acid biosynthesis, was used to examine the effects of fatty acids on replication of VZV and posttranslational processing of glycoproteins. Treatment of the cells with cerulenin significantly inhibited viral growth, but did not affect protein synthesis in the cells. Examination of processing of viral glycoproteins … More in treated cells revealed the accumulation of the precursor proteins, and decrease in the amount of mature glycoproteins. these data suggest that fatty acid acylation of VZV glycoproteins is necessary for formation of complete infectious virions. Next, plasmid DNAs encoding gpI,gpII, gpIII and gpIV were inseted into vaccinia virus DNA and the recombinant viruses were used for immunological analysis. In consequence, the gpI derived from cells infected with virus showed have very similar antigenic characteristics as gpI derived from VZV infected cells by immunoprecipitation pattern, and antibody produced in rabbits infected with recombinant virus had high neutralizing activity, when the reaction was performed with complement. However, no virus specific polypeptides were detected in cells infected with other recombinant viruses, although RNAs were detected in infected cells. This result suggested that gpI has important role for protection and recovery from VZV infection. Finally, specificity of the skin test with VZV glycoprotein was examined in guinea pigs infected with herpes simplex virus(HSV) type 1 and VZV and in children with a history of HSV infection who developed varicella. No cross-reaction between HSV and VZV was detected and the skin test was specific for immunity to VZV infection in children. These results suggested that VZV skin test is specific for immunity to VZV infection. This specificity will be of value in screening or immunity to VZV, irrespective of prior HSV infection. Less

水痘-带状疱疹病毒（VZV）含有四种主要的糖蛋白，命名为gpI、gpII、gpIII和gpIV。首先，检查这些糖蛋白中脂肪酸的存在和作用。用[^3H]-棕榈酸标记的感染细胞研究了脂肪酸与VZV跨膜糖蛋白的结合。用抗VZV糖蛋白的单克隆抗体处理细胞提取物，并用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）分析沉淀物。电泳图谱显示gpI被重标记，而gpII和IV被轻标记。浅蓝菌素，一种抑制从头脂肪酸生物合成的抗生素，用于检查脂肪酸对VZV复制和糖蛋白翻译后加工的影响。用浅蓝菌素处理细胞显著抑制病毒生长，但不影响细胞中的蛋白质合成。病毒糖蛋白加工的检查 ...更多信息在处理的细胞中，显示了前体蛋白的积累和成熟糖蛋白的量的减少。这些数据表明VZV糖蛋白的脂肪酸酰化对于完整感染性病毒体的形成是必需的。接着，将编码gpI、gpII、gpIII和gpIV的质粒DNA插入到牛痘病毒DNA中，并将重组病毒用于免疫学分析。结果，通过免疫沉淀模式，来自病毒感染细胞的gpI显示出与来自VZV感染细胞的gpI具有非常相似的抗原特性，并且当与补体进行反应时，在重组病毒感染的兔中产生的抗体具有高中和活性。然而，在感染其他重组病毒的细胞中未检测到病毒特异性多肽，尽管在感染的细胞中检测到RNA。提示gpI在VZV感染后的保护和恢复过程中具有重要作用。最后，在单纯疱疹病毒（HSV）1型和VZV感染的豚鼠和患有水痘的HSV感染史的儿童中检查了VZV糖蛋白皮肤试验的特异性。HSV与VZV之间无交叉反应，对儿童VZV感染具有特异性。提示VZV皮试具有特异性。这种特异性将在筛查或VZV免疫中具有价值，而与既往HSV感染无关。少