Hemphill M L, Forng R Y, Abernathy E S, Frey T K
Department of Biology, Georgia State University, Atlanta 30303.
Virology. 1988 Jan;162(1):65-75. doi: 10.1016/0042-6822(88)90395-9.
Virus specific macromolecular synthesis was studied in Vero cells infected with plaque-purified rubella virus under one-step multiplication conditions. Under these conditions, the rate of virus production was found to increase rapidly until 24 hr postinfection after which time the rate of virus production rose more slowly, reaching a peak level at 48 hr postinfection. This peak rate of virus production was maintained through 72 hr postinfection. A majority of the cells remained alive through 96 hr postinfection, although a 20 to 30% decrease in the number of living cells occurred between 24 and 48 hr postinfection, the time period at which cytopathic effect was first observed. The virus structural proteins were first detected intracellularly at 16 hr postinfection. The rate of synthesis of these proteins was already maximal at 16 hr postinfection and remained constant through 48 hr postinfection. By immunofluorescence, cells expressing virus proteins were first observed at 12 hr postinfection. At 24 hr postinfection, 35 to 50% of the cells in the infected culture were exhibiting immunofluorescence, at 36 hr postinfection, 65 to 90% of the cells were exhibiting immunofluorescence, and at 48 hr postinfection, all of the cells were exhibiting immunofluorescence. The virus genomic and subgenomic RNA species were first detectable by 12 hr postinfection. The rate of synthesis of both of these species peaked at 26 hr postinfection. Rubella virus infection was found to have no effect on total cell RNA synthesis. However, a modest inhibition of total cell protein synthesis which reached 40% by 48 hr postinfection was observed. When Northern analysis of RNA extracted from infected cells was performed, a negative-polarity, virus-specific RNA probe hybridized only to the virus genomic and subgenomic RNA species. A positive-polarity, virus-specific RNA probe hybridized predominantly to a negative-polarity RNA of genome length indicating that both the genomic and subgenomic RNAs are synthesized from a genome-length negative-polarity template. Defective interfering (DI) RNAs were not detected in infected cells through 96 hr postinfection or in cells onto which virus released through 96 hr postinfection was passaged. Thus, the generation of DI particles by rubella virus appears to play no role in the slow, noncytopathic replication of this virus or in the ability of rubella virus-infected cells to survive for extended periods of time.
在一步增殖条件下，对感染蚀斑纯化风疹病毒的非洲绿猴肾细胞（Vero细胞）中的病毒特异性大分子合成进行了研究。在这些条件下，发现病毒产生速率在感染后24小时内迅速增加，此后病毒产生速率上升较慢，在感染后48小时达到峰值水平。病毒产生的峰值速率在感染后72小时内保持不变。大多数细胞在感染后96小时内仍存活，尽管在感染后24至48小时之间活细胞数量减少了20%至30%，这是首次观察到细胞病变效应的时间段。病毒结构蛋白在感染后16小时首次在细胞内被检测到。这些蛋白的合成速率在感染后16小时已达到最大值，并在感染后48小时内保持恒定。通过免疫荧光法，在感染后12小时首次观察到表达病毒蛋白的细胞。在感染后24小时，感染培养物中35%至50%的细胞呈现免疫荧光，在感染后36小时，65%至90%的细胞呈现免疫荧光，在感染后48小时，所有细胞都呈现免疫荧光。病毒基因组和亚基因组RNA种类在感染后12小时首次可检测到。这两种RNA的合成速率在感染后26小时达到峰值。发现风疹病毒感染对细胞总RNA合成没有影响。然而，观察到细胞总蛋白合成受到适度抑制，在感染后48小时达到40%。当对从感染细胞中提取的RNA进行Northern分析时，负极性病毒特异性RNA探针仅与病毒基因组和亚基因组RNA种类杂交。正极性病毒特异性RNA探针主要与基因组长度的负极性RNA杂交，表明基因组和亚基因组RNA均由基因组长度的负极性模板合成。在感染后96小时内的感染细胞中或在接种感染后96小时释放的病毒的细胞中未检测到缺陷干扰（DI）RNA。因此，风疹病毒产生DI颗粒似乎在该病毒缓慢的非细胞病变复制或风疹病毒感染细胞长时间存活的能力中不起作用。
