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感染火鸡肠道冠状病毒的HRT - 18细胞的超微结构及蛋白A - 金免疫标记

Ultrastructure and protein A-gold immunolabelling of HRT-18 cells infected with turkey enteric coronavirus.

作者信息

Dea S, Garzon S, Strykowski H, Tijssen P

机构信息

Centre de Recherche en Médecine Comparée, Institut Armand-Frappier, Université du Québec, Laval-des-Rapides, Canada.

出版信息

Vet Microbiol. 1989 May;20(1):21-33. doi: 10.1016/0378-1135(89)90004-7.

Abstract

The Minnesota strain of turkey enteric coronavirus (TCV) was propagated in HRT-18 cells, a cell line derived from human rectum adenocarcinoma. A productive non-cytopathic infection was established, without a previous adaptation, in these cells as shown by the specific hemagglutinating activity in cell culture supernatants. A post-embedding immunochemical technique, using specific antiserum directed against the original egg-adapted virus and colloidal-gold-labelled protein A as the electron-dense marker, was used for the identification of the virus and related antigens in the cells by electron microscopy. Budding of typical coronavirus particles, through intracytoplasmic membranes and accumulation of complete virus within cytoplasmic vesicles or the lumen of rough endoplasmic reticulum, were the main features of the viral morphogenesis. Late in infection, numerous progeny viral particles were shown at the outer surface of infected cells, but budding could not be demonstrated at this level. Two different types of surface projections were observed on the extracellular particles of this avian coronavirus. These morphological characteristics have been thus far described only for mammalian hemagglutinating coronaviruses.

摘要

火鸡肠道冠状病毒(TCV)的明尼苏达毒株在HRT - 18细胞中繁殖,HRT - 18细胞系源自人直肠腺癌。如细胞培养上清液中的特异性血凝活性所示,在这些细胞中未预先适应就建立了有效的非细胞病变感染。采用包埋后免疫化学技术,使用针对原始鸡胚适应病毒的特异性抗血清和胶体金标记的蛋白A作为电子致密标记物,通过电子显微镜鉴定细胞中的病毒及相关抗原。典型冠状病毒颗粒通过胞质内膜出芽以及完整病毒在胞质囊泡或粗面内质网腔内积累,是病毒形态发生的主要特征。感染后期,在受感染细胞的外表面可见大量子代病毒颗粒,但在此水平无法证明出芽现象。在这种禽冠状病毒的细胞外颗粒上观察到两种不同类型的表面突起。迄今为止,这些形态特征仅在哺乳动物血凝冠状病毒中有所描述。

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