Ye Li, Wang Xu, Wang Shihong, Luo Guangxiang, Wang Yanjian, Liang Hao, Ho Wenzhe
Division of Allergy & Immunology, Joseph Stokes, Jr. Research Institute at The Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
J Virol Methods. 2008 Mar;148(1-2):161-5. doi: 10.1016/j.jviromet.2007.11.002.
Hepatitis C virus (HCV) is a human pathogen associated with chronic liver disease. Recently, the cell culture systems supporting complete replication and production of HCV genotype 2a (JFH1) have been established. This study investigated the effect of low-speed centrifugation on HCV JFH1 infection of human hepatocytes (Huh7.5.1). Higher levels of HCV RNA expression were observed in Huh7.5.1 cells infected with centrifugal inoculation of HCV JFH1 than those in the control cells. This increased HCV RNA expression was associated with the elevated expression of HCV NS3 protein in the hepatocytes. The centrifugal enhancement of HCV infection was time and speed dependent. However, the enhancement was not observed when centrifugation was performed before or after HCV infection. In addition, there was no association between centrifugal enhancement and the expression of HCV entry receptors (CD81 and claudin-1) and intracellular IFN-alpha in the hepatocytes. These data indicate that centrifugal inoculation is a useful tool for increasing the efficiency of HCV infection and replication in the target cells in vitro.
丙型肝炎病毒（HCV）是一种与慢性肝病相关的人类病原体。最近，支持丙型肝炎病毒2a基因型（JFH1）完全复制和产生的细胞培养系统已经建立。本研究调查了低速离心对人肝细胞（Huh7.5.1）丙型肝炎病毒JFH1感染的影响。与对照细胞相比，经离心接种丙型肝炎病毒JFH1感染的Huh7.5.1细胞中观察到更高水平的丙型肝炎病毒RNA表达。这种丙型肝炎病毒RNA表达的增加与肝细胞中丙型肝炎病毒NS3蛋白表达的升高有关。丙型肝炎病毒感染的离心增强作用具有时间和速度依赖性。然而，在丙型肝炎病毒感染之前或之后进行离心时未观察到增强作用。此外，离心增强与肝细胞中丙型肝炎病毒进入受体（CD81和claudin-1）的表达以及细胞内干扰素-α之间没有关联。这些数据表明，离心接种是一种提高丙型肝炎病毒在体外靶细胞中感染和复制效率的有用工具。
