A hepatocellular carcinoma cell line producing mature hepatitis B viral particles.
Current in vitro models for hepatitis B virus (HBV) are based on human hepatoblastoma cell lines transfected with HBV genome. The objective of this work was to develop an in vitro, hepatocellular carcinoma (HCC)-based system supporting HBV full replication and producing mature viral particles. The FLC4 human HCC cell line was stably transfected with a plasmid carrying a head-to-tail dimer of the adwHBV genome. One of the clones, FLC4A10II, exhibited prolonged expression of HBV, as was demonstrated by secreted levels of HBsAg, HBeAg, and HBV DNA in the culture medium of the growing cells. Furthermore, the cells produced HBV particles that were detected by a cesium chloride density gradient performed on the culture medium. Analysis by Southern blot revealed that HBV DNA has integrated into the FLC4A10II cell genome. The presence of HBV in the FLC4A10II cells did not cause alterations in cell morphology and the cells continued to resemble mature hepatocytes. They do exhibit a high mitotic activity. The new HBV stably transfected cell line, FLC4A10II, can serve as an important tool for further exploration of HBV host-pathogen interaction, viral life cycle, and for assessing new antiviral agents.
Fellig
Yakov
Y
Department of Pathology, Hadassah University Hospital, Jerusalem, Israel.
Almogy
Gidon
G
Galun
Eithan
E
Ketzinel-Gilad
Mali
M
D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000821:virology*; D045744:Cell Line, Tumor; D003470:Culture Media; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D019065:Virus Assembly; D016662:Virus Integration
D003470:Culture Media; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D012367:RNA, Viral; D014764:Viral Proteins