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

作　　者：Yingying Song Shuyu Shou Huimin Guo Zixiang Gao Nannan Liu Yang Yang Feifei Wang Qiang Deng Jing Liu Youhua Xie 

机构地区：[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]Institute for Hepatology,National Clinical Research Center for Infectious Disease,Shenzhen Third People's Hospital,Shenzhen,518112,China [3]Children's Hospital,Fudan University,Shanghai,201102,China [4]Shanghai Key Laboratory of Medical Epigenetics,Institutes of Biomedical Sciences,School of Basic Medical Sciences,Shanghai Medical College,Fudan University,Shanghai,200032,China [5]The Second Affiliated Hospital,School of Medicine,Southern University of Science and Technology,Shenzhen,518112,China

出　　处：《Virologica Sinica》2022年第4期558-568,共11页中国病毒学（英文版）

基　　金：the National Natural Science Foundation of China(81971921,81971931);the National base cultivation project(20DZ2210404);the Major science and technology project for the prevention and treatment of major infectious diseases(2018ZX10301208)。

摘　　要：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(pg RNA)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 HNF4αand human sodium taurocholate cotransporting polypeptide(h NTCP)expression into BEL7404made 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 h NTCP and Tet-inducible HNF4αfollowed 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.

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

分 类 号：R373.21[医药卫生—病原生物学]