Hill C L, Booth T F, Stuart D I, Mertens P P
NERC Institute of Virology and Environmental Microbiology, Oxford, UK.
J Virol Methods. 1999 Mar;78(1-2):177-89. doi: 10.1016/s0166-0934(98)00181-5.
Cytoplasmic polyhedrosis viruses (CPV) are classified as 14 distinct species (electropherotypes) within the genus Cypovirus, family Reoviridae. Cypovirus research has been limited by a lack of appropriate cell culture systems (for each of these virus species) in which the majority of cells can become productively infected. Lipofection increased the infection rate of Lymantria dispar 652 cells, by virus particles (derived from polyhedra) of Orgyia pseudosugata type 5 cypovirus (Op-5 CPV), from 3 to 44%. Lipofection also significantly increased the percentage of Trichoplusia ni 368 cells infected with the same virus (from < 1 to approximately 7%). The spread of cypovirus infection between cells was either very slow or insignificant, and infected cells appeared to remain viable for long periods. Virus infection was detected by the observation of polyhedra formation in individual cells and it was therefore possible to develop a simple quantitative assay system to measure virus titre (TCID50). Cryo-electron microscopy showed that cypovirus particles formed a complex with the lipid, involving their envelopment within the liposome membrane. It was concluded that the increased infectivity of the virus by lipofection was due to a more efficient cell entry mechanism, probably involving fusion between liposome and cell membranes.
细胞质多角体病毒(CPV)在呼肠孤病毒科质型多角体病毒属内被分为14个不同的种(电泳型)。质型多角体病毒的研究一直受到缺乏合适细胞培养系统(针对这些病毒的每个种)的限制,在这种系统中大多数细胞能够被有效感染。脂质体转染将舞毒蛾652细胞被云杉毒蛾5型质型多角体病毒(Op-5 CPV)的病毒粒子(从多角体中提取)感染的比例从3%提高到了44%。脂质体转染还显著提高了被同一种病毒感染的粉纹夜蛾368细胞的比例(从小于1%提高到了约7%)。质型多角体病毒在细胞间的传播非常缓慢或不明显,且被感染的细胞似乎能长时间保持存活。通过观察单个细胞中多角体的形成来检测病毒感染,因此有可能开发一种简单的定量检测系统来测量病毒滴度(半数组织培养感染剂量)。冷冻电子显微镜显示质型多角体病毒粒子与脂质形成了复合物,包括它们被包裹在脂质体膜内。得出的结论是,脂质体转染使病毒感染性增加是由于更有效的细胞进入机制,可能涉及脂质体与细胞膜之间的融合。