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狂犬病病毒在C6大鼠胶质瘤细胞(克隆CCL-107)中的分离与增殖

Isolation and replication of rabies virus in C6 rat glioma cells (clone CCL-107).

作者信息

Bordignon J, Piza A T, Alvarez-Silva M, Caporale G M, Carrieri M L, Kotait I, Zanetti C R

机构信息

Department of Microbiology, Immunology and Parasitology of the Federal University of Santa Catarina (MIP/CCB/UFSC), Campus Universitário da Trindade, Florianópolis, 88040-900, Brazil.

出版信息

Biologicals. 2001 Jun;29(2):67-73. doi: 10.1006/biol.2001.0278.

Abstract

The susceptibility of the C6 rat glioma cell line (ATCC; CCL-107) to rabies virus was characterized. The kinetics of infection performed with a fixed and a wild strain (from an infected cow) of rabies virus was monitored by direct immunofluorescence. Fluorescent cytoplasmic bodies were readily observed by UV microscopy from 24 hours post-infection (hpi) onwards. The ability of C6 to produce rabies infective virion particles was confirmed by determining the viral titres present in the supernatants of infected cultures, by both BHK-21 cell infection and mice inoculation. C6 cells produced similar viral titres to those produced by BHK-21 for both strains used. In addition, the yield of rabies glycoprotein was assessed by ELISA. In general, BHK-21 and C6 cells infected either by PV or with the wild rabies strain produced similar amounts of rabies glycoprotein. At 96 hpi, however, when the glycoprotein production peaked, BHK-21 infected with the wild strain produced significantly higher amounts of glycoprotein than C6. Subsequently, the optimal conditions for isolation of wild rabies virus strains from C6 cells were established and these proved to be as sensitive as NA cells in detecting 10 wild rabies samples. Due to the high sensitivity exhibited, C6 rat glioma cells present a new and useful system for rabies virus investigation.

摘要

对C6大鼠胶质瘤细胞系(美国典型培养物保藏中心;CCL - 107)对狂犬病病毒的易感性进行了表征。通过直接免疫荧光监测了用狂犬病病毒的固定毒株和野生毒株(来自一头感染牛)进行感染的动力学。从感染后24小时(hpi)起,通过紫外线显微镜很容易观察到荧光细胞质体。通过BHK - 21细胞感染和小鼠接种来测定感染培养物上清液中的病毒滴度,从而证实了C6产生狂犬病感染性病毒粒子的能力。对于所使用的两种毒株,C6细胞产生的病毒滴度与BHK - 21产生的相似。此外,通过酶联免疫吸附测定法评估狂犬病糖蛋白的产量。一般来说,用PV或野生狂犬病毒株感染的BHK - 21和C6细胞产生的狂犬病糖蛋白量相似。然而,在96 hpi时,当糖蛋白产量达到峰值时,用野生毒株感染的BHK - 21产生的糖蛋白量明显高于C6。随后,确定了从C6细胞中分离野生狂犬病病毒毒株的最佳条件,并且这些条件在检测10个野生狂犬病样本时被证明与NA细胞一样敏感。由于表现出的高敏感性,C6大鼠胶质瘤细胞为狂犬病病毒研究提供了一个新的有用系统。

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