A robust cell culture system supporting the complete life cycle of hepatitis B virus

The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as the hepatitis B virus (HBV) receptor enabled researchers to create hepatoma cell lines susceptible to HBV infection. Infection in current systems, however, is inefficient and virus fails to spread. Infection efficiency is enha...

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Autores principales: Michailidis, Eleftherios, Pabon, Jonathan, Xiang, Kuanhui, Park, Paul, Ramanan, Vyas, Hoffmann, Hans-Heinrich, Schneider, William M., Bhatia, Sangeeta N., de Jong, Ype P., Shlomai, Amir, Rice, Charles M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
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Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5709435/
https://www.ncbi.nlm.nih.gov/pubmed/29192196
http://dx.doi.org/10.1038/s41598-017-16882-5
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author Michailidis, Eleftherios
Pabon, Jonathan
Xiang, Kuanhui
Park, Paul
Ramanan, Vyas
Hoffmann, Hans-Heinrich
Schneider, William M.
Bhatia, Sangeeta N.
de Jong, Ype P.
Shlomai, Amir
Rice, Charles M.
author_facet Michailidis, Eleftherios
Pabon, Jonathan
Xiang, Kuanhui
Park, Paul
Ramanan, Vyas
Hoffmann, Hans-Heinrich
Schneider, William M.
Bhatia, Sangeeta N.
de Jong, Ype P.
Shlomai, Amir
Rice, Charles M.
author_sort Michailidis, Eleftherios
collection PubMed
description The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as the hepatitis B virus (HBV) receptor enabled researchers to create hepatoma cell lines susceptible to HBV infection. Infection in current systems, however, is inefficient and virus fails to spread. Infection efficiency is enhanced by treating cells with polyethylene glycol 8000 (PEG) during infection. However, this alone does not promote virus spread. Here we show that maintaining PEG in culture medium increases the rate of infection by at least one order of magnitude, and, most importantly, promotes virus spread. To demonstrate the utility of this system, we show that two interferon-stimulated genes (ISGs), ISG20 and tetherin, restrict HBV spread in NTCP-expressing hepatoma cells. Thus, this protocol can be easily applied to existing cell culture systems to study the complete HBV life cycle, including virus spread.
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spelling pubmed-57094352017-12-06 A robust cell culture system supporting the complete life cycle of hepatitis B virus Michailidis, Eleftherios Pabon, Jonathan Xiang, Kuanhui Park, Paul Ramanan, Vyas Hoffmann, Hans-Heinrich Schneider, William M. Bhatia, Sangeeta N. de Jong, Ype P. Shlomai, Amir Rice, Charles M. Sci Rep Article The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as the hepatitis B virus (HBV) receptor enabled researchers to create hepatoma cell lines susceptible to HBV infection. Infection in current systems, however, is inefficient and virus fails to spread. Infection efficiency is enhanced by treating cells with polyethylene glycol 8000 (PEG) during infection. However, this alone does not promote virus spread. Here we show that maintaining PEG in culture medium increases the rate of infection by at least one order of magnitude, and, most importantly, promotes virus spread. To demonstrate the utility of this system, we show that two interferon-stimulated genes (ISGs), ISG20 and tetherin, restrict HBV spread in NTCP-expressing hepatoma cells. Thus, this protocol can be easily applied to existing cell culture systems to study the complete HBV life cycle, including virus spread. Nature Publishing Group UK 2017-11-30 /pmc/articles/PMC5709435/ /pubmed/29192196 http://dx.doi.org/10.1038/s41598-017-16882-5 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Michailidis, Eleftherios
Pabon, Jonathan
Xiang, Kuanhui
Park, Paul
Ramanan, Vyas
Hoffmann, Hans-Heinrich
Schneider, William M.
Bhatia, Sangeeta N.
de Jong, Ype P.
Shlomai, Amir
Rice, Charles M.
A robust cell culture system supporting the complete life cycle of hepatitis B virus
title A robust cell culture system supporting the complete life cycle of hepatitis B virus
title_full A robust cell culture system supporting the complete life cycle of hepatitis B virus
title_fullStr A robust cell culture system supporting the complete life cycle of hepatitis B virus
title_full_unstemmed A robust cell culture system supporting the complete life cycle of hepatitis B virus
title_short A robust cell culture system supporting the complete life cycle of hepatitis B virus
title_sort robust cell culture system supporting the complete life cycle of hepatitis b virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5709435/
https://www.ncbi.nlm.nih.gov/pubmed/29192196
http://dx.doi.org/10.1038/s41598-017-16882-5
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