Monoclonal antibodies against three major glycoproteins of varicella-zoster virus.
Monoclonal antibodies against three major glycoproteins of varicella-zoster virus.
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针对水痘带状疱疹病毒三种主要糖蛋白的单克隆抗体。
DOI：
10.1128/iai.40.1.381-388.1983
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发表时间：
1983
影响因子：
3.1
通讯作者：
McGuire,WL
中科院分区：
文献类型：
--
作者：
Grose,C;Edwards,DP;Friedrichs,WE;Weigle,KA;McGuire,WL
水痘带状疱疹病毒（VZV）在受感染的细胞培养物中编码三种主要的糖蛋白——gp62、gp98和gp118。为了分别表征这些已知的免疫原，我们首先用VZV粗提取物接种BALB/c小鼠，通过Köhler-Milstein细胞融合技术产生杂杂瘤培养物，并通过间接免疫荧光筛选培养上清，以检测针对未固定VZV感染细胞的反应性（FAMA测定）。5个独立衍生和亚克隆的VZV- fama滴度高但对未感染或单纯疱疹病毒感染的细胞无反应性的杂交瘤上清进一步通过[3H]聚焦标记和清洁剂溶解的VZV抗原制剂进行免疫沉淀分析。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对沉淀进行分离，发现4种单克隆抗体能与gp62和gp98反应，1种单克隆抗体只与gp118反应。无论在还原或非还原条件下，剖面都没有变化。当检测中和活性时，单个抗gp118杂交瘤的分泌产物，而不是四个抗gp62/gp98克隆的上清液，抑制VZV斑块形成超过80%。因此，FAMA试验检测到的糖基化抗原中至少有一个是引起中和活性的决定因素。
Varicella-zoster virus (VZV) codes for three prominent glycoproteins--gp62, gp98, and gp118--in infected cell cultures. To characterize individually these known immunogens, we first inoculated BALB/c mice with crude VZV extracts, produced hybridoma cultures by Köhler-Milstein cell-fusion technology, and screened culture supernatants by indirect immunofluorescence for reactivity directed against unfixed VZV-infected cells (FAMA assay). Supernatants from five independently derived and subcloned hybridomas with a high VZV-FAMA titer but no reactivity against either uninfected or herpes simplex virus-infected cells were further analyzed by immunoprecipitation of [3H]fucose-labeled and detergent-solubilized VZV antigen preparations. Fractionation of the precipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that four monoclonal antibodies reacted with both gp62 and gp98, and one precipitated only gp118. The profiles were unchanged whether performed under reducing or nonreducing conditions. When assayed for neutralizing activity, the secretory product of the single anti-gp118 hybridoma, but not the supernatants from the four anti-gp62/gp98 clones, inhibited VZV plaque formation by greater than 80%. Thus, at least one of the glycosylated antigens detected by the FAMA assay is a determinant which elicits neutralizing activity.