标题: Entry of Hepatitis B Virus into Immortalized Human Primary Hepatocytes by Clathr [打印本页] 作者: StephenW 时间: 2012-8-10 14:28 标题: Entry of Hepatitis B Virus into Immortalized Human Primary Hepatocytes by Clathr
Hsiu-Chen Huang a,
Chun-Chi Chen b,
Wen-Cheng Chang c,
Mi-Hua Tao d and
Cheng Huang c
- Author Affiliations
a Department of Applied Science, National Hsinchu University of Education, Hsinchu, Taiwan
b CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
c National Research Institute of Chinese Medicine, Taipei, Taiwan
d Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan
ABSTRACT
The lack of a suitable in vitro hepatitis B virus (HBV) infectivity model has limited examination of the early stages of the virus-cell interaction. In this study, we used an immortalized cell line derived from human primary hepatocytes, HuS-E/2, to study the mechanism of HBV infection. HBV infection efficiency was markedly increased after dimethyl sulfoxide (DMSO)-induced differentiation of the cells. Transmission electron microscopy demonstrated the presence of intact HBV particles in DMSO-treated HBV-infected HuS-E/2 cells, which could be infected with HBV for up to at least 50 passages. The pre-S1 domain of the large HBsAg (LHBsAg) protein specifically interacted with clathrin heavy chain (CHC) and clathrin adaptor protein AP-2. Short hairpin RNA knockdown of CHC or AP-2 in HuS-E/2 cells significantly reduced their susceptibility to HBV, indicating that both are necessary for HBV infection. Furthermore, HBV entry was inhibited by chlorpromazine, an inhibitor of clathrin-mediated endocytosis. LHBsAg also interfered with the clathrin-mediated endocytosis of transferrin by human hepatocytes. This infection system using an immortalized human primary hepatocyte cell line will facilitate investigations into HBV entry and in devising therapeutic strategies for manipulating HBV-associated liver disorders. 作者: StephenW 时间: 2012-8-10 14:29