Vincent S, Spehner D, Manié S, Delorme R, Drillien R, Gerlier D
Immunité Infections Virales, IVMC, CNRS-UCBL UMR 5537, Lyon Cedex 08, 69372, France.
Virology. 1999 Dec 20;265(2):185-95. doi: 10.1006/viro.1999.0064.
Infection of mouse L.CD46 fibroblasts with measles virus resulted in a poor virus yield, although no defects in the steps of virus binding, entry or fusion, were detected. Two days post-infection, the level of expression of the viral F protein was found to be similar on the surface of infected L.CD46 and HeLa cells using a virus multiplicity enabling an equal number of cells to be infected. After immunofluorescence labelling and confocal microscopy, L.CD46 cells also displayed a significant increase in the co-localisation of the N protein with the cell surface H and F proteins. Immunogold labelling and transmission electron microscopy demonstrated the accumulation of numerous nucleocapsids near the plasma membrane of L. CD46 cells with little virus budding, in contrast to infected HeLa cells which displayed fewer cortical nucleocapsids and more enveloped viral particles. Purified virus particles from infected L. CD46 contained a reduced amount of H, F and M protein. Altogether, these data indicate that, in L.CD46 cells, the late stage of measles virus assembly is defective. This cellular model will be helpful for the identification of cellular factors controlling measles virus maturation.
用麻疹病毒感染小鼠L.CD46成纤维细胞，尽管在病毒结合、进入或融合步骤中未检测到缺陷，但病毒产量很低。感染后两天，使用能使相同数量细胞被感染的病毒复数，发现在被感染的L.CD46细胞和HeLa细胞表面，病毒F蛋白的表达水平相似。经过免疫荧光标记和共聚焦显微镜观察，L.CD46细胞中N蛋白与细胞表面H蛋白和F蛋白的共定位也显著增加。免疫金标记和透射电子显微镜显示，与感染的HeLa细胞相比，L.CD46细胞的质膜附近积累了大量核衣壳，病毒出芽很少，而HeLa细胞的皮质核衣壳较少，包膜病毒颗粒较多。从感染的L.CD46中纯化的病毒颗粒所含的H蛋白、F蛋白和M蛋白量减少。总之，这些数据表明，在L.CD46细胞中，麻疹病毒组装的后期存在缺陷。这种细胞模型将有助于鉴定控制麻疹病毒成熟的细胞因子。
