Abstract
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, FLC4A10 II, 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 FLC4A10 II cell genome. The presence of HBV in the FLC4A10 II 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, FLC4A10 II, can serve as an important tool for further exploration of HBV host-pathogen interaction, viral life cycle, and for assessing new antiviral agents.
Original language | English |
---|---|
Pages (from-to) | 269-274 |
Number of pages | 6 |
Journal | Biochemical and Biophysical Research Communications |
Volume | 321 |
Issue number | 2 |
DOIs | |
State | Published - 20 Aug 2004 |
Bibliographical note
Funding Information:We thank Ruth Adler, Rima Barsuk, and Athalia Klein (Liver Unit, Hadassah Hospital) for technical help and ongoing discussions; Orit Pappo and Naomi Ron (Pathology Department, Hadassah hospital) for their insights and technical support; and Reba Condiotti and Hilla Giladi (Goldyne Savad Institute of Gene Therapy, Hadassah hospital) for critical reading of the manuscript. We thank Prof. Tatsuo Miyamura and Yoichiro Aoki from the National Institute of Infectious Diseases, Tokyo, Japan, for providing us FLC4 cells. This study was supported by the Blum and Grinspoon foundation and by the Young Clinicians Foundation (Hadassah Organization).
Keywords
- HBV
- Hepatocellular carcinoma cell lines