Ng M-L, Tan S-H, See E-E, Ooi E-E, Ling A-E
Department of Microbiology, 5 Science Drive 2, National University of Singapore, 117597, Singapore.
Electron Microscopy Unit, Faculty of Medicine, 5 Science Drive 2, National University of Singapore, 117597, Singapore.
J Gen Virol. 2003 Dec;84(Pt 12):3291-3303. doi: 10.1099/vir.0.19505-0.
An isolate of SARS coronavirus (strain 2003VA2774) was obtained from a patient and used to infect Vero E6 cells. The replication cycle of the virus was followed from 1 to 30 h post-infection (p.i.). It was surprising to observe the swift growth of this human virus in Vero cells. Within the first hour of infection, the most obvious ultrastructural change was the proliferation of the Golgi complexes and related vesicles accompanied by swelling of some of the trans-Golgi sacs. Extracellular virus particles were present by 5 h p.i. in about 5 % of the cells and this increased dramatically to about 30 % of the cell population within an hour (6 h p.i.). Swollen Golgi sacs contained virus nucleocapsids at different stages of maturation. These virus precursors were also in large vacuoles and in close association with membrane whorls. The membrane whorls could be the replication complexes, since they appeared rather early in the replication cycle. As infection progressed from 12 to 21 h p.i., the cytoplasm of the infected cells was filled with numerous large, smooth-membraned vacuoles containing a mixture of mature virus and spherical cores. Several of these vacuoles were close to the cell periphery, ready to export out the mature progeny virus particles via exocytosis. By 24 to 30 h p.i., crystalline arrays of the extracellular virus particles were seen commonly at the cell surface.
从一名患者身上分离出一株严重急性呼吸综合征冠状病毒(毒株2003VA2774),并用其感染Vero E6细胞。在感染后1至30小时(p.i.)跟踪病毒的复制周期。令人惊讶的是,这种人类病毒在Vero细胞中迅速生长。在感染的第一个小时内,最明显的超微结构变化是高尔基体复合体和相关囊泡的增殖,同时一些反式高尔基体囊泡肿胀。在感染后5小时,约5%的细胞中出现了细胞外病毒颗粒,在一小时内(感染后6小时),这一比例急剧增加至约30%的细胞群体。肿胀的高尔基体囊泡中含有处于不同成熟阶段的病毒核衣壳。这些病毒前体也存在于大液泡中,并与膜性漩涡紧密相连。膜性漩涡可能是复制复合体,因为它们在复制周期中出现得相当早。随着感染从感染后12小时进展到21小时,被感染细胞的细胞质中充满了大量含有成熟病毒和球形核心混合物的大的、光滑膜性液泡。其中一些液泡靠近细胞周边,准备通过胞吐作用输出成熟的子代病毒颗粒。到感染后24至30小时,在细胞表面通常可以看到细胞外病毒颗粒的晶体阵列。