Booth T F, Gould E A, Nuttall P A
NERC Institute of Virology and Environmental Microbiology, Oxford, U.K.
Virus Res. 1991 Nov;21(3):199-212. doi: 10.1016/0168-1702(91)90033-r.
We have studied the structure and morphogenesis of Dugbe (DUG) virus (Bunyaviridae, Nairovirus) in cultured porcine kidney (PS) cells and a tick cell line (Ra 243) using immunogold electron microscopy. DUG virus is a tickborne arbovirus, considered to be a low health hazard, that is antigenically and genetically related to Crimean Congo haemorrhagic fever (CCHF) virus (Marriott et al., 1990). We have investigated the maturation and intracellular transport of DUG virus particles as a model for other more pathogenic nairoviruses using monoclonal antibodies for immunogold labelling of ultrathin cryosections and immunofluorescence techniques. The spherical DUG virus particle measures about 90 nm in diameter, with a 5 nm thick membrane covered by 5-7 nm long projections or "spikes". These projections form hollow cylindrical morphological units, about 5 nm in diameter. DUG virus infection caused only a slight cytopathogenic effect in mammalian cells and none in tick cells. DUG virus particles assembled by budding from the Golgi complex, where the DUG virus glycoprotein G1 accumulated in vesicles originating from Golgi cisternae. The nucleocapsid protein N accumulated in scattered foci throughout the cytoplasm, and this appears to be related to the limited maturation of DUG virus particles that occurred. The reduced number of budding virus particles observed in tick cells was correlated with the reduced cytopathology observed.
我们利用免疫金电子显微镜技术,研究了杜格(DUG)病毒(布尼亚病毒科,内罗毕病毒属)在猪肾(PS)细胞培养物和蜱细胞系(Ra 243)中的结构和形态发生。DUG病毒是一种蜱传虫媒病毒,被认为对健康危害较小,在抗原性和基因上与克里米亚刚果出血热(CCHF)病毒相关(Marriott等人,1990年)。我们使用单克隆抗体对超薄冷冻切片进行免疫金标记和免疫荧光技术,以DUG病毒颗粒的成熟和细胞内运输作为其他更具致病性的内罗毕病毒的模型进行了研究。球形的DUG病毒颗粒直径约为90纳米,有一层5纳米厚的膜,上面覆盖着5至7纳米长的突起或“刺突”。这些突起形成直径约5纳米的中空圆柱形形态单位。DUG病毒感染在哺乳动物细胞中仅引起轻微的细胞病变效应,在蜱细胞中则无此效应。DUG病毒颗粒通过从高尔基体复合体出芽组装而成,其中DUG病毒糖蛋白G1在源自高尔基体池的小泡中积累。核衣壳蛋白N在整个细胞质中散在的病灶中积累,这似乎与发生的DUG病毒颗粒有限的成熟有关。在蜱细胞中观察到的出芽病毒颗粒数量减少与观察到的细胞病变减少相关。