2. Academic Validation
  3. Establishment and characterization of a new cell culture system for hepatitis B virus replication and infection

Establishment and characterization of a new cell culture system for hepatitis B virus replication and infection

  • Virol Sin. 2022 Aug;37(4):558-568. doi: 10.1016/j.virs.2022.05.002.
Yingying Song 1,  Shuyu Shou 1,  Huimin Guo 2,  Zixiang Gao 1,  Nannan Liu 1,  Yang Yang 1,  Feifei Wang 1,  Qiang Deng 1,  Jing Liu 3,  Youhua Xie 4
Affiliations

Affiliations

  • 1 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Institute of Infectious Diseases and Biosecurity, Shanghai Medical College, Fudan University, Shanghai, 200032, China.
  • 2 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Institute of Infectious Diseases and Biosecurity, Shanghai Medical College, Fudan University, Shanghai, 200032, China; Institute for Hepatology, National Clinical Research Center for Infectious Disease, Shenzhen Third People's Hospital, Shenzhen, 518112, China; The Second Affiliated Hospital, School of Medicine, Southern University of Science and Technology, Shenzhen, 518112, China.
  • 3 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Institute of Infectious Diseases and Biosecurity, Shanghai Medical College, Fudan University, Shanghai, 200032, China; Shanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, 200032, China. Electronic address: liujing212@fudan.edu.cn.
  • 4 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Institute of Infectious Diseases and Biosecurity, Shanghai Medical College, Fudan University, Shanghai, 200032, China; Children's Hospital, Fudan University, Shanghai, 201102, China. Electronic address: yhxie@fudan.edu.cn.
PMID: 35568375 DOI: 10.1016/j.virs.2022.05.002
Abstract

Hepatitis B virus (HBV) is a primary cause of chronic liver diseases in humans. HBV Infection exhibits strict host and tissue tropism. HBV core promoter (Cp) drives transcription of pregenomic RNA (pgRNA) and plays a key role in the viral life cycle. Hepatocyte nuclear factor 4α (HNF4α) acts as a major transcriptional factor that stimulates Cp. In this work, we reported that BEL7404 ​cell line displayed a high efficiency of DNA transfection and high levels of HBV antigen expression after transfection of HBV replicons without prominent viral replication. The introduction of exogenous HNF4α and human sodium taurocholate cotransporting polypeptide (hNTCP) expression into BEL7404 made it permissive for HBV replication and susceptible to HBV Infection. BEL7404-derived cell lines with induced HBV permissiveness and susceptibility were constructed by stable co-transfection of hNTCP and Tet-inducible HNF4α followed by limiting dilution cloning. HBV replication in such cells was sensitive to inhibition by nucleotide analog tenofovir, while the Infection was inhibited by HBV entry inhibitors. This Cell Culture system provides a new and additional tool for the study of HBV replication and Infection as well as the characterization of Antiviral agents.

Keywords

BEL7404; Doxycycline (DOX); Hepatitis B virus (HBV); Hepatocyte nuclear factor 4α (HNF4α); Sodium-taurocholate cotransporting polypeptide (NTCP); Tetracycline (Tet)-On.

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