Vieyres Gabrielle, Angus Allan G N, Haberstroh Anita, Baumert Thomas F, Dubuisson Jean, Patel Arvind H
MRC Virology Unit, Institute of Virology, University of Glasgow, Glasgow, UK.
J Virol Methods. 2009 Apr;157(1):69-79. doi: 10.1016/j.jviromet.2008.11.015. Epub 2009 Jan 7.
Hepatitis C virus (HCV) entry into target cells is thought to be a multistep process involving several cellular factors. However, their precise role during virus entry is unclear. Investigation of the mechanisms of HCV entry, such as the order of intervention by the cellular receptors, requires synchronizing infections. This study describes a new method involving magnetic adsorption of virus to nanoparticles to synchronize infection, which can be adapted to both HCV pseudoparticles and cell culture infectious HCV. By combining these particles with negatively or positively charged magnetic nanoparticles it was possible to adsorb them onto target cells under a magnetic field in only 2min. This resulted in greater efficiency of virus adsorption to cells, and increased the infectivity of cell culture infectious virus, as compared to the standard protocol involving incubation of the virus with cells at 4 degrees C for 1h, or to a standard infection at 37 degrees C. Furthermore, magnetic adsorption respected the natural entry route of the virus, making this system suitable to study the early stages of HCV infection.
丙型肝炎病毒(HCV)进入靶细胞被认为是一个涉及多种细胞因子的多步骤过程。然而,它们在病毒进入过程中的精确作用尚不清楚。对HCV进入机制的研究,如细胞受体干预的顺序,需要同步感染。本研究描述了一种新方法,该方法涉及通过病毒与纳米颗粒的磁性吸附来同步感染,这一方法可适用于HCV假颗粒和细胞培养感染性HCV。通过将这些颗粒与带负电或正电的磁性纳米颗粒相结合,能够在磁场作用下仅用2分钟就将它们吸附到靶细胞上。与在4℃下将病毒与细胞孵育1小时的标准方案或在37℃下的标准感染相比,这导致病毒吸附到细胞上的效率更高,并提高了细胞培养感染性病毒的感染性。此外,磁性吸附遵循病毒的自然进入途径,使得该系统适合于研究HCV感染的早期阶段。