Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus

Abstract Human hepatitis delta virus (HDV) causes the most severe form of viral hepatitis. Approximately 15–25 million people are chronically infected with HDV. As a satellite virus of the human hepatitis B virus (HBV), HDV uses the HBV-encoded envelope proteins for egress from and de novo entry int...

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Autores principales:	Yi Ni, Zhenfeng Zhang, Lisa Engelskircher, Georg Verch, Thomas Tu, Florian A. Lempp, Stephan Urban
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/30ccad5c8a38486fb94b4a7814b23893
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spelling	oai:doaj.org-article:30ccad5c8a38486fb94b4a7814b238932021-12-02T15:09:20ZGeneration and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus10.1038/s41598-019-46493-12045-2322https://doaj.org/article/30ccad5c8a38486fb94b4a7814b238932019-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-46493-1https://doaj.org/toc/2045-2322Abstract Human hepatitis delta virus (HDV) causes the most severe form of viral hepatitis. Approximately 15–25 million people are chronically infected with HDV. As a satellite virus of the human hepatitis B virus (HBV), HDV uses the HBV-encoded envelope proteins for egress from and de novo entry into hepatocytes. So far, in vitro production of HDV particles is restricted to co-transfection of cells with HDV/HBV encoding cDNAs. This approach has several limitations. In this study, we established HuH7-END cells, which continuously secrete infectious HDV virions. The cell line was generated through stepwise stable integration of the cDNA of the HDV antigenome, the genes for the HBV envelope proteins and the HBV/HDV receptor NTCP. We found that HuH7-END cells release infectious HDV particles up to 400 million copies/milliliter and support virus spread to co-cultured cells. Due to the expression of NTCP, HuH7-END cells are also susceptible to de novo HDV entry. Virus production is stable for >16 passages and can be scaled up for preparation of large HDV virus stocks. Finally, HuH7-END cells are suitable for screening of antiviral drugs targeting HDV replication. In summary, the HuH7-END cell line provides a novel tool to study HDV replication in vitro.Yi NiZhenfeng ZhangLisa EngelskircherGeorg VerchThomas TuFlorian A. LemppStephan UrbanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-14 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yi Ni Zhenfeng Zhang Lisa Engelskircher Georg Verch Thomas Tu Florian A. Lempp Stephan Urban Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
description	Abstract Human hepatitis delta virus (HDV) causes the most severe form of viral hepatitis. Approximately 15–25 million people are chronically infected with HDV. As a satellite virus of the human hepatitis B virus (HBV), HDV uses the HBV-encoded envelope proteins for egress from and de novo entry into hepatocytes. So far, in vitro production of HDV particles is restricted to co-transfection of cells with HDV/HBV encoding cDNAs. This approach has several limitations. In this study, we established HuH7-END cells, which continuously secrete infectious HDV virions. The cell line was generated through stepwise stable integration of the cDNA of the HDV antigenome, the genes for the HBV envelope proteins and the HBV/HDV receptor NTCP. We found that HuH7-END cells release infectious HDV particles up to 400 million copies/milliliter and support virus spread to co-cultured cells. Due to the expression of NTCP, HuH7-END cells are also susceptible to de novo HDV entry. Virus production is stable for >16 passages and can be scaled up for preparation of large HDV virus stocks. Finally, HuH7-END cells are suitable for screening of antiviral drugs targeting HDV replication. In summary, the HuH7-END cell line provides a novel tool to study HDV replication in vitro.
format	article
author	Yi Ni Zhenfeng Zhang Lisa Engelskircher Georg Verch Thomas Tu Florian A. Lempp Stephan Urban
author_facet	Yi Ni Zhenfeng Zhang Lisa Engelskircher Georg Verch Thomas Tu Florian A. Lempp Stephan Urban
author_sort	Yi Ni
title	Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
title_short	Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
title_full	Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
title_fullStr	Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
title_full_unstemmed	Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
title_sort	generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/30ccad5c8a38486fb94b4a7814b23893
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Generation and characterization of a stable cell line persistently replicating and secreting the human hepatitis delta virus

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