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HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection

HepG2 cells reconstituted with Hepatitis B virus (HBV) entry receptor sodium taurocholate co-transporting polypeptide (NTCP) are widely used as a convenient in vitro cell culture infection model for HBV replication studies. As such, it is pertinent that HBV infectivity is maintained at steady-state...

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Autores principales: Zahoor, Muhammad Atif, Kuipery, Adrian, Mosa, Alexander I., Gehring, Adam J., Feld, Jordan J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9412438/
https://www.ncbi.nlm.nih.gov/pubmed/36016422
http://dx.doi.org/10.3390/v14081800
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author Zahoor, Muhammad Atif
Kuipery, Adrian
Mosa, Alexander I.
Gehring, Adam J.
Feld, Jordan J.
author_facet Zahoor, Muhammad Atif
Kuipery, Adrian
Mosa, Alexander I.
Gehring, Adam J.
Feld, Jordan J.
author_sort Zahoor, Muhammad Atif
collection PubMed
description HepG2 cells reconstituted with Hepatitis B virus (HBV) entry receptor sodium taurocholate co-transporting polypeptide (NTCP) are widely used as a convenient in vitro cell culture infection model for HBV replication studies. As such, it is pertinent that HBV infectivity is maintained at steady-state levels for an accurate interpretation of in vitro data. However, variations in the HBV infection efficiency due to imbalanced NTCP expression levels in the HepG2 cell line may affect experimental results. In this study, we performed single cell-cloning of HepG2-NTCP-A3 parental cells via limiting dilution and obtained multiple subclones with increased permissiveness to HBV. Specifically, one subclone (HepG2-NTCP-A3/C2) yielded more than four-fold higher HBV infection compared to the HepG2-NTCP-A3 parental clone. In addition, though HBV infectivity was universally reduced in the absence of polyethylene glycol (PEG), subclone C2 maintained relatively greater permissiveness under PEG-free conditions, suggesting the functional heterogeneity within parental HepG2-NTCP-A3 may be exploitable in developing a PEG-free HBV infection model. The increased viral production correlated with increased intracellular viral antigen expression as evidenced through HBcAg immunofluorescence staining. Further, these subclones were found to express different levels of NTCP, albeit with no remarkable morphology or cell growth differences. In conclusion, we isolated the subclones of HepG2-NTCP-A3 which support efficient HBV production and thus provide an improved in vitro HBV infection model.
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spelling pubmed-94124382022-08-27 HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection Zahoor, Muhammad Atif Kuipery, Adrian Mosa, Alexander I. Gehring, Adam J. Feld, Jordan J. Viruses Brief Report HepG2 cells reconstituted with Hepatitis B virus (HBV) entry receptor sodium taurocholate co-transporting polypeptide (NTCP) are widely used as a convenient in vitro cell culture infection model for HBV replication studies. As such, it is pertinent that HBV infectivity is maintained at steady-state levels for an accurate interpretation of in vitro data. However, variations in the HBV infection efficiency due to imbalanced NTCP expression levels in the HepG2 cell line may affect experimental results. In this study, we performed single cell-cloning of HepG2-NTCP-A3 parental cells via limiting dilution and obtained multiple subclones with increased permissiveness to HBV. Specifically, one subclone (HepG2-NTCP-A3/C2) yielded more than four-fold higher HBV infection compared to the HepG2-NTCP-A3 parental clone. In addition, though HBV infectivity was universally reduced in the absence of polyethylene glycol (PEG), subclone C2 maintained relatively greater permissiveness under PEG-free conditions, suggesting the functional heterogeneity within parental HepG2-NTCP-A3 may be exploitable in developing a PEG-free HBV infection model. The increased viral production correlated with increased intracellular viral antigen expression as evidenced through HBcAg immunofluorescence staining. Further, these subclones were found to express different levels of NTCP, albeit with no remarkable morphology or cell growth differences. In conclusion, we isolated the subclones of HepG2-NTCP-A3 which support efficient HBV production and thus provide an improved in vitro HBV infection model. MDPI 2022-08-17 /pmc/articles/PMC9412438/ /pubmed/36016422 http://dx.doi.org/10.3390/v14081800 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Brief Report
Zahoor, Muhammad Atif
Kuipery, Adrian
Mosa, Alexander I.
Gehring, Adam J.
Feld, Jordan J.
HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection
title HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection
title_full HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection
title_fullStr HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection
title_full_unstemmed HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection
title_short HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection
title_sort hepg2-ntcp subclones exhibiting high susceptibility to hepatitis b virus infection
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9412438/
https://www.ncbi.nlm.nih.gov/pubmed/36016422
http://dx.doi.org/10.3390/v14081800
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