Ogino Michiko, Yoshimatsu Kumiko, Ebihara Hideki, Araki Koichi, Lee Byoung-Hee, Okumura Megumi, Arikawa Jiro
Institute for Animal Experimentation, Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
J Virol. 2004 Oct;78(19):10776-82. doi: 10.1128/JVI.78.19.10776-10782.2004.
Hantaan virus (HTNV)-infected Vero E6 cells undergo cell fusion with both infected and uninfected cells under low-pH conditions. Flow cytometry and fluorescence microscopy of HTNV-infected Vero E6 cells showed that envelope glycoproteins (GPs) were located both on the cell surface and in the cytoplasm. Neutralizing monoclonal antibodies (MAbs) against the G1 and G2 envelope GPs inhibited cell fusion, whereas nonneutralizing MAbs against G1 or G2 and MAbs against the nucleocapsid protein (NP) did not. Transfected Vero E6 cells that expressed GPs but not those that expressed NP fused and formed syncytia. These results indicate that HTNV GPs act as fusogens at the cell surface. No fusion activity was observed either in infected Vero cells that were passaged more than 150 times or in BHK-21 cells, although GPs appeared to localize to the cell surface. This variability in fusion induction suggests the involvement of host cell factors in the process of cell membrane fusion.
汉坦病毒(HTNV)感染的Vero E6细胞在低pH条件下会与感染和未感染的细胞发生细胞融合。对HTNV感染的Vero E6细胞进行流式细胞术和荧光显微镜观察发现,包膜糖蛋白(GPs)既位于细胞表面,也存在于细胞质中。针对G1和G2包膜糖蛋白的中和单克隆抗体(MAbs)可抑制细胞融合,而针对G1或G2的非中和MAbs以及针对核衣壳蛋白(NP)的MAbs则不能。转染表达GPs的Vero E6细胞会发生融合并形成多核巨细胞,而表达NP的细胞则不会。这些结果表明,HTNV糖蛋白在细胞表面充当融合原。在传代超过150次的感染Vero细胞或BHK - 21细胞中均未观察到融合活性,尽管糖蛋白似乎定位于细胞表面。这种融合诱导的变异性表明宿主细胞因子参与了细胞膜融合过程。