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dc.contributor.authorTodt, Daniel
dc.contributor.authorFriesland, Martina
dc.contributor.authorMoeller, Nora
dc.contributor.authorPraditya, Dimas
dc.contributor.authorKinast, Volker
dc.contributor.authorBrüggemann, Yannick
dc.contributor.authorKnegendorf, Leonard
dc.contributor.authorBurkard, Thomas
dc.contributor.authorSteinmann, Joerg
dc.contributor.authorBurm, Rani
dc.contributor.authorVerhoye, Lieven
dc.contributor.authorWahid, Avista
dc.contributor.authorMeister, Toni Luise
dc.contributor.authorEngelmann, Michael
dc.contributor.authorPfankuche, Vanessa M
dc.contributor.authorPuff, Christina
dc.contributor.authorVondran, Florian W R
dc.contributor.authorBaumgärtner, Wolfgang
dc.contributor.authorMeuleman, Philip
dc.contributor.authorBehrendt, Patrick
dc.contributor.authorSteinmann, Eike
dc.date.accessioned2020-01-15T13:52:03Z
dc.date.available2020-01-15T13:52:03Z
dc.date.issued2020-01-02
dc.identifier.citationProc Natl Acad Sci U S A. 2020 Jan 2. pii: 1912307117. doi: 10.1073/pnas.1912307117.en_US
dc.identifier.issn1091-6490
dc.identifier.pmid31896581
dc.identifier.doi10.1073/pnas.1912307117
dc.identifier.urihttp://hdl.handle.net/10033/622084
dc.description.abstractHepatitis E virus (HEV) is the causative agent of hepatitis E in humans and the leading cause for acute viral hepatitis worldwide. The virus is classified as a member of the genus Orthohepevirus A within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle, the development of effective antivirals and a vaccine is severely limited. In this study, we established a protocol based on the HEV genotype 3 p6 (Kernow C-1) and the human hepatoma cell lines HepG2 and HepG2/C3A with different media conditions to produce intracellular HEV cell culture-derived particles (HEVcc) with viral titers between 105 and 106 FFU/mL. Viral titers could be further enhanced by an HEV variant harboring a mutation in the RNA-dependent RNA polymerase. These HEVcc particles were characterized in density gradients and allowed the trans-complementation of subgenomic reporter HEV replicons. In addition, in vitro produced intracellular-derived particles were infectious in liver-humanized mice with high RNA copy numbers detectable in serum and feces. Efficient infection of primary human and swine hepatocytes using the developed protocol could be observed and was inhibited by ribavirin. Finally, RNA sequencing studies of HEV-infected primary human hepatocytes demonstrated a temporally structured transcriptional defense response. In conclusion, this robust cell culture model of HEV infection provides a powerful tool for studying viral-host interactions that should facilitate the discovery of antiviral drugs for this important zoonotic pathogen.en_US
dc.language.isoenen_US
dc.publisherNational Academy of Sciencesen_US
dc.relationinfo:eu-repo/grantAgreement/EC/H2020/643476en_US
dc.rightsopenAccessen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjecthepatitis E virus (HEV)en_US
dc.subjecthumanized miceen_US
dc.subjectinfectionen_US
dc.subjectprimary hepatocytesen_US
dc.subjecttranscriptomicsen_US
dc.titleRobust hepatitis E virus infection and transcriptional response in human hepatocytes.en_US
dc.typeArticleen_US
dc.contributor.departmentTWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.en_US
dc.identifier.journalProceedings of the National Academy of sciencesen_US
dc.source.journaltitleProceedings of the National Academy of Sciences of the United States of America


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