Pizzato M, Marlow S A, Blair E D, Takeuchi Y
Chester Beatty Laboratories, Institute of Cancer Research, London SW3 6JB, United Kingdom.
J Virol. 1999 Oct;73(10):8599-611. doi: 10.1128/JVI.73.10.8599-8611.1999.
The initial step of virus-cell interaction was studied by immunofluorescence microscopy. Single particles of murine leukemia virus (MLV) vectors and human immunodeficiency virus (HIV) were visualized by immunofluorescence. Fluorescent dots representing single virions could be localized by staining of capsid proteins (CA) or surface envelope proteins (SU) after fixation of virus supernatants. This technique can be used to determine particle concentration in viral supernatants and also to study virus-cell interaction. We investigated the role of the Env-receptor interaction for the initial binding event between the cell and the viral particles. Ecotropic MLV vector particles were shown to bind to human cells which do not express the specific viral receptor. In addition, MLV particles defective for Env were shown to bind the cells similarly to infectious MLV. Time course experiments of virus-cell binding and dissociation showed identical profiles for infectious and Env-defective MLV particles and suggested that MLV Env is not involved in the early phases of attachment of virus to cells. The possible implication of cellular factors in enhancing viral binding and infectivity is discussed.
通过免疫荧光显微镜研究了病毒与细胞相互作用的初始步骤。通过免疫荧光观察到了鼠白血病病毒(MLV)载体和人类免疫缺陷病毒(HIV)的单个颗粒。在固定病毒上清液后,通过对衣壳蛋白(CA)或表面包膜蛋白(SU)进行染色,可以定位代表单个病毒粒子的荧光点。该技术可用于确定病毒上清液中的颗粒浓度,还可用于研究病毒与细胞的相互作用。我们研究了Env受体相互作用在细胞与病毒颗粒之间初始结合事件中的作用。嗜亲性MLV载体颗粒被证明可与不表达特定病毒受体的人类细胞结合。此外,Env缺陷型MLV颗粒被证明与感染性MLV一样能与细胞结合。病毒与细胞结合和解离的时间进程实验表明,感染性和Env缺陷型MLV颗粒具有相同的特征,这表明MLV Env不参与病毒附着于细胞的早期阶段。文中还讨论了细胞因子在增强病毒结合和感染性方面的可能影响。