Shiraki K, Takahashi M
J Gen Virol. 1982 Aug;61 (Pt 2):271-5. doi: 10.1099/0022-1317-61-2-271.
Virus particles and virus proteins excreted from cultured human embryonic lung (HEL) cells infected with varicella-zoster virus (VZV) were examined by electron microscopy and affinity column chromatography using an antibody to VZV followed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Approximately 1 X 10(9) to 2 X 10(9) virus particles/ml with no detectable infectivity, of which 30 to 80% were enveloped, were observed in the culture fluid 48 to 72 h after infection, when cytopathic effect (c.p.e.) appeared. In the sonicated infected cell suspension, 1 X 10(9) to 2 X 10(9) virus particles/ml, of which 30 to 50% were enveloped, were observed and the virus particle/infectivity ratio was approx. 10(6):1. The culture fluid of infected HEL cells labelled with [35S]methionine or [3H]glucosamine was centrifuged at 1000000 g for 2 h to remove virus particles and the supernatant was examined for excreted virus proteins. Affinity column chromatography of the supernatant using immobilized human zoster convalescent serum, led to the isolation of virus antigens which were analyzed by SDS-PAGE. Polypeptides with mol. wt. of approx. 115K and 45K, both of which were glycosylated, were detected, suggesting that these VZV glycoproteins were excreted from the infected cells.
运用电子显微镜以及亲和柱层析法（先使用水痘-带状疱疹病毒（VZV）抗体，随后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）），对感染水痘-带状疱疹病毒（VZV）的人胚肺（HEL）细胞培养物中排出的病毒颗粒和病毒蛋白进行了检测。感染后48至72小时，当出现细胞病变效应（c.p.e.）时，在培养液中观察到约1×10⁹至2×10⁹个/毫升无明显感染性的病毒颗粒，其中30%至80%有包膜。在超声处理的感染细胞悬液中，观察到1×10⁹至2×10⁹个/毫升的病毒颗粒，其中30%至50%有包膜，病毒颗粒与感染性的比例约为10⁶:1。用[³⁵S]甲硫氨酸或[³H]葡糖胺标记的感染HEL细胞的培养液在1000000g下离心2小时以去除病毒颗粒，然后检测上清液中的排出病毒蛋白。使用固定化的人带状疱疹康复血清对上清液进行亲和柱层析，分离出病毒抗原，并通过SDS-PAGE进行分析。检测到分子量约为115K和45K的两种糖基化多肽，表明这些VZV糖蛋白是从感染细胞中排出的。
