RESCUE OF A POSITIVE STRANDED RNA VIRUS FROM ANTIGEN NEGATIVE NEURO-BLASTOMA CELLS
RESCUE OF A POSITIVE STRANDED RNA VIRUS FROM ANTIGEN NEGATIVE NEURO-BLASTOMA CELLS
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DOI：
10.1016/0024-3205(79)90323-0
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发表时间：
1979-01-01
期刊：
影响因子：
6.1
通讯作者：
WEINER, LP
中科院分区：
文献类型：
--
作者：
STOHLMAN, SA;SAKAGUCHI, AY;WEINER, LP
潜伏感染的小鼠神经母细胞瘤细胞的单细胞克隆分离自在抗病毒抗体存在下用小鼠肝炎病毒慢性感染的培养物。这些细胞克隆在32、37或39 ℃培养期间不产生感染性病毒或显示病毒性细胞病变效应。C.感染性病毒通过使用聚乙二醇与允许细胞融合从单细胞克隆中分离，但在与灭活仙台病毒融合后或用代谢抑制剂处理后未分离。经免疫荧光检测，其中一个拯救病毒的细胞克隆（S-3）的病毒抗原呈阴性。通过使用细胞质提取物进行的补体结合试验或通过聚丙烯酰胺凝胶电泳检测的病毒特异性蛋白，S-3细胞克隆没有可证实的病毒抗原。拯救的病毒在32 ℃时表现出温度依赖性生长缺陷。C和被归类为冷敏感突变体。正链RNA病毒的完整基因组可以在感染细胞中保持潜伏，而不表达可检测的病毒抗原。
Single cell clones of latently infected mouse neuroblastoma cells were isolated from a culture chronically infected with mouse hepatitis virus in the presence of an antiviral antibody. These cell clones did not produce infectious virus or exhibit viral cytopathic effects during cultivation at 32, 37 or 39.degree. C. Infectious virus was isolated from single cell clones via fusion with permissive cells using polyethylene glycol, but not after fusion with inactivated Sendai virus or following treatment with metabolic inhibitors. One cell clone (S-3) from which virus was rescued was negative for viral antigen by immunofluorescence. The S-3 cell clone had no demonstrable virus antigen by complement-fixation tests using cytoplasmic extracts or virus-specified proteins detectable by polyacrylamide gel electrophoresis. The rescued viruses exhibited a temperature dependent growth defect at 32.degree. C and were classified as cold sensitive mutants. A complete genome of a positive stranded RNA virus can remain latent in infected cells without the expression of detectable virus antigen.