Hughes J H
Department of Medical Microbiology & Immunology, Ohio State University, Columbus 43210.
Clin Microbiol Rev. 1993 Apr;6(2):150-75. doi: 10.1128/CMR.6.2.150.
Viral replication events can be enhanced by physical, chemical, or heat treatment of cells. The centrifugation of cells can stimulate them to proliferate, reduce their generation times, and activate gene expression. Human endothelial cells can be activated to release cyclo-oxygenase metabolites after rocking for 5 min, and mechanical stress can stimulate endothelial cells to proliferate. Centrifugation of virus-infected cultures can increase cytopathic effects (CPE), enhance the number of infected cells, increase viral yields, and reduce viral detection times and may increase viral isolation rates. The rolling of virus-infected cells also has an effect similar to that of centrifugation. The continuous rolling of virus-infected cultures at < or = 2.0 rpm can enhance enterovirus, rhinovirus, reovirus, rotavirus, paramyxovirus, herpesvirus, and vaccinia virus CPE or yields or both. For some viruses, the continuous rolling of infected cell cultures at 96 rpm (1.9 x g) is superior to rolling at 2.0 rpm for viral replication or CPE production. In addition to centrifugation and rolling, the treatment of cells with chemicals or heat can also enhance viral yields or CPE. For example, the treatment of virus-infected cells with dimethyl sulfoxide can enhance viral transformation, increase plaque numbers and plaque size, increase the number of cells producing antigens, and increase viral yields. The infectivity of fowl plague virus is increased by 80-fold when 4% dimethyl sulfoxide is added to culture medium immediately after infection. The heat shocking of virus-infected cells also has been shown to have a stimulatory effect on the replication events of cytomegalovirus, Epstein-Barr virus, and human immunodeficiency virus. The effects of motion, chemicals, or heat treatments on viral replication are not well understood. These treatments apparently activate cells to make them more permissive to viral infection and viral replication. Perhaps heat shock proteins or stress proteins are a common factor for this enhancement phenomenon. The utility of these treatments alone or in combination with other methods for enhancing viral isolation and replication in a diagnostic setting needs further investigation.
病毒复制事件可通过对细胞进行物理、化学或热处理来增强。细胞离心可刺激其增殖、缩短其代时并激活基因表达。人内皮细胞在摇晃5分钟后可被激活以释放环氧化酶代谢产物，机械应力可刺激内皮细胞增殖。对病毒感染的培养物进行离心可增加细胞病变效应（CPE）、增加感染细胞数量、提高病毒产量、缩短病毒检测时间并可能提高病毒分离率。病毒感染细胞的滚动也具有与离心类似的效果。以≤2.0转/分钟的速度持续滚动病毒感染的培养物可增强肠道病毒、鼻病毒、呼肠孤病毒、轮状病毒、副粘病毒、疱疹病毒和痘苗病毒的CPE或产量或两者。对于某些病毒，以96转/分钟（1.9×g）的速度持续滚动感染细胞培养物在病毒复制或CPE产生方面优于以2.0转/分钟的速度滚动。除了离心和滚动外，用化学物质或热处理细胞也可提高病毒产量或CPE。例如，用二甲基亚砜处理病毒感染的细胞可增强病毒转化、增加蚀斑数量和蚀斑大小、增加产生抗原的细胞数量并提高病毒产量。感染后立即向培养基中添加4%二甲基亚砜时，禽瘟病毒的感染性可提高80倍。对病毒感染的细胞进行热休克也已显示对巨细胞病毒、爱泼斯坦-巴尔病毒和人类免疫缺陷病毒的复制事件具有刺激作用。运动、化学物质或热处理对病毒复制的影响尚不清楚。这些处理显然激活细胞，使其对病毒感染和病毒复制更具易感性。也许热休克蛋白或应激蛋白是这种增强现象的共同因素。这些处理单独使用或与其他方法联合用于在诊断环境中增强病毒分离和复制的效用需要进一步研究。
