Entry of Hepatitis B Virus into Immortalized Human Primary Hepatocytes by Clathr

StephenW

http://jvi.asm.org/content/86/17/9443.abstract?etoc
Entry of Hepatitis B Virus into Immortalized Human Primary Hepatocytes by Clathrin-Dependent Endocytosis

    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.

StephenW

B型肝炎病毒进入永生化人原代肝细胞网格蛋白依赖的内吞作用

    修陈黄A，
    陈骏池B，
    张文成C，
    陶秘华d和
    郑黄C

- 作者背景

    应用科学系，国立新竹教育大学，台湾新竹
    b中国科学院病原微生物学与免疫学重点实验室，中国科学院微生物研究所，中国科学院，北京，中国
    3C国家研究中国医药研究所，台北，台湾
    ð，台北，台湾中央研究院生物医学科学研究所

摘要

适合在体外乙型肝炎病毒（HBV）感染性模型的缺乏限制了病毒细胞相互作用的早期阶段的考试。在这项研究中，我们使用来自人类原发性肝细胞，HUS-/ 2的永生细胞株，研究乙肝病毒感染的机制。二甲基亚砜（DMSO）诱导分化的细胞后，HBV感染效率显着增加。透射电镜显示完整的乙肝病毒颗粒的存在，在二甲基亚砜治疗乙肝病毒感染的HUS E /2细胞，这可能是与乙肝病毒感染，至少50代。专门网格蛋白重链（CHC）的网格接头蛋白AP-2相互作用的大型乙肝表面抗原（LHBsAg）蛋白前S1域。文委会或AP-2的短发夹RNA击倒HUS E /2细胞显着减少其对HBV的易感性，表明这两个是乙肝病毒感染的必要条件。此外，氯丙嗪，网格蛋白介导的内吞作用的抑制剂抑制乙肝病毒进入。 LHBsAg也干扰了网格蛋白介导的转人肝细胞的内吞作用。这种感染系统使用一个永生化人原发性肝细胞株，将促进乙肝病毒入境，并在制定操纵HBV相关的肝脏疾病的治疗策略的调查。

MP4

台湾國立中國醫藥研究所美女黃琤
还有牛人陶秘华和北京中科院。