Proteomic Analysis of HepaRG Cells: A Novel Cell Line That Supports Hepatitis B Virus Infection

Narayan, Raj; Gangadharan, Bevin; Hantz, Olivier; Antrobus, Robin; García, Ángel; Dwek, Raymond A.; Zitzmann, Nicole

doi:10.1021/pr800562j

Cited by 28 publications

(23 citation statements)

References 41 publications

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“…Basic markers for diagnosis of HBV infection include the presence of hepatitis B surface antigens (HBsAgs) and hepatitis B envelope antigens in acute or chronically infected hepatocytes [4,5]. Several physiological and biochemical methods have been developed to monitor HBV infection [6,7,8,9]. In addition, Abe et al reported quantitative analysis of HBV using DNA Polymerase chain reaction (PCR) assay [10].…”

Section: Introductionmentioning

confidence: 99%

Development of Lateral Flow Assay Based on Size-Controlled Gold Nanoparticles for Detection of Hepatitis B Surface Antigen

Kim

Hong

et al. 2016

Sensors

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In this study, we developed lateral flow assay (LFA) biosensors for the detection of hepatitis B surface antigens using well-controlled gold nanoparticles (AuNPs). To enhance colorimetric signals, a seeded growth method was used for the preparation of size-controlled AuNPs with a narrow size distribution. Different sizes of AuNPs in the range of 342–137.8 nm were conjugated with antibodies and then optimized for the efficient detection of LFA biosensors. The conjugation stability was investigated by UV-vis spectroscopy of AuNP dispersion at various pH values and concentrations of antibody. Based on optimized conjugation conditions, the use of 42.7 ± 0.8 nm AuNPs exhibited superior performance for the detection of LFAs relative to other sizes of AuNPs.

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Section: Introductionmentioning

confidence: 99%

Development of Lateral Flow Assay Based on Size-Controlled Gold Nanoparticles for Detection of Hepatitis B Surface Antigen

Kim

Hong

et al. 2016

Sensors

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“…A previous proteomics study using HBV-uninfected and HBV-infected HepaRG cells identified 19 differentially-regulated proteins [12]. However, additional proteomic studies, more focused on plasma membrane proteins, (the first recognition partners during cell-virus interaction), are needed.…”

Section: Introductionmentioning

confidence: 99%

Proteomic analysis of plasma membranes isolated from undifferentiated and differentiated HepaRG cells

et al. 2012

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Liver infection with hepatitis B virus (HBV), a DNA virus of the Hepadnaviridae family, leads to severe disease, such as fibrosis, cirrhosis and hepatocellular carcinoma. The early steps of the viral life cycle are largely obscure and the host cell plasma membrane receptors are not known. HepaRG is the only proliferating cell line supporting HBV infection in vitro, following specific differentiation, allowing for investigation of new host host-cell factors involved in viral entry, within a more robust and reproducible environment. Viral infection generally begins with receptor recognition at the host cell surface, following highly specific cell-virus interactions. Most of these interactions are expected to take place at the plasma membrane of the HepaRG cells. In the present study, we used this cell line to explore changes between the plasma membrane of undifferentiated (−) and differentiated (+) cells and to identify differentially-regulated proteins or signaling networks that might potentially be involved in HBV entry. Our initial study identified a series of proteins that are differentially expressed in the plasma membrane of (−) and (+) cells and are good candidates for potential cell-virus interactions. To our knowledge, this is the first study using functional proteomics to study plasma membrane proteins from HepaRG cells, providing a platform for future experiments that will allow us to understand the cell-virus interaction and mechanism of HBV viral infection.

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“…To investigate the complete life cycle of HBV infection—despite the usefulness of available human liver cell lines—there is clearly a strong need for appropriate in vitro infection systems, allowing for exogenous HBV infection [7]. HBV replication can be initiated by genomic DNA transfection into HepG2, Huh7, HepAD38, or primary hepatocytes [8]. However, until recent years, exogenous HBV infection has only been successfully achieved using primary human hepatocytes [9], [10].…”

Section: Introductionmentioning

confidence: 99%

An Occult Hepatitis B-Derived Hepatoma Cell Line Carrying Persistent Nuclear Viral DNA and Permissive for Exogenous Hepatitis B Virus Infection

et al. 2013

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Occult hepatitis B virus (HBV) infection is defined as persistence of HBV DNA in liver tissues, with or without detectability of HBV DNA in the serum, in individuals with negative serum HBV surface antigen (HBsAg). Despite accumulating evidence suggesting its important clinical roles, the molecular and virological basis of occult hepatitis B remains unclear. In an attempt to establish new hepatoma cell lines, we achieved a new cell line derived from a hepatoma patient with chronic hepatitis C virus (HCV) and occult HBV infection. Characterization of this cell line revealed previously unrecognized properties. Two novel human hepatoma cell lines were established. Hep-Y1 was derived from a male hepatoma patient negative for HCV and HBV infection. Hep-Y2 was derived from a female hepatoma patient suffering from chronic HCV and occult HBV infection. Morphological, cytogenetic and functional studies were performed. Permissiveness to HBV infection was assessed. Both cell lines showed typical hepatocyte-like morphology under phase-contrast and electron microscopy and expressed alpha-fetoprotein, albumin, transferrin, and aldolase B. Cytogenetic analysis revealed extensive chromosomal anomalies. An extrachromosomal form of HBV DNA persisted in the nuclear fraction of Hep-Y2 cells, while no HBsAg was detected in the medium. After treated with 2% dimethyl sulfoxide, both cell lines were permissive for exogenous HBV infection with transient elevation of the replication intermediates in the cytosol with detectable viral antigens by immunoflurescence analysis. In conclusions, we established two new hepatoma cell lines including one from occult HBV infection (Hep-Y2). Both cell lines were permissive for HBV infection. Additionally, Hep-Y2 cells carried persistent extrachromosomal HBV DNA in the nuclei. This cell line could serve as a useful tool to establish the molecular and virological basis of occult HBV infection.

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Proteomic Analysis of HepaRG Cells: A Novel Cell Line That Supports Hepatitis B Virus Infection

Cited by 28 publications

References 41 publications

Development of Lateral Flow Assay Based on Size-Controlled Gold Nanoparticles for Detection of Hepatitis B Surface Antigen

Development of Lateral Flow Assay Based on Size-Controlled Gold Nanoparticles for Detection of Hepatitis B Surface Antigen

Proteomic analysis of plasma membranes isolated from undifferentiated and differentiated HepaRG cells

An Occult Hepatitis B-Derived Hepatoma Cell Line Carrying Persistent Nuclear Viral DNA and Permissive for Exogenous Hepatitis B Virus Infection

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