A hepatocellular carcinoma cell line producing mature hepatitis B viral particles

Yakov Fellig; Gidon Almogy; Eithan Galun; Mali Ketzinel-Gilad

doi:10.1016/j.bbrc.2004.06.148

A hepatocellular carcinoma cell line producing mature hepatitis B viral particles

Yakov Fellig, Gidon Almogy, Eithan Galun, Mali Ketzinel-Gilad

Faculty of Medicine

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

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	https://doi.org/10.1016/j.bbrc.2004.06.148
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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bbrc.2004.06.148

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title = "A hepatocellular carcinoma cell line producing mature hepatitis B viral particles",

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.",

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author = "Yakov Fellig and Gidon Almogy and Eithan Galun and Mali Ketzinel-Gilad",

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).",

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AU - Almogy, Gidon

AU - Galun, Eithan

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N2 - 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.

AB - 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.

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A hepatocellular carcinoma cell line producing mature hepatitis B viral particles

Abstract

Bibliographical note

Keywords

UN SDGs

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