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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2020,1,21]]},"abstract":"<jats:p>\n                    Hepatitis 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\n                    <jats:italic>Orthohepevirus A<\/jats:italic>\n                    within the\n                    <jats:italic>Hepeviridae<\/jats:italic>\n                    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 10\n                    <jats:sup>5<\/jats:sup>\n                    and 10\n                    <jats:sup>6<\/jats:sup>\n                    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\n                    <jats:italic>trans<\/jats:italic>\n                    -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\u2013host interactions that should facilitate the discovery of antiviral drugs for this important zoonotic pathogen.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1912307117","type":"journal-article","created":{"date-parts":[[2020,1,2]],"date-time":"2020-01-02T20:06:04Z","timestamp":1577995564000},"page":"1731-1741","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":121,"title":["Robust hepatitis E virus infection and transcriptional response in human hepatocytes"],"prefix":"10.1073","volume":"117","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3564-1014","authenticated-orcid":false,"given":"Daniel","family":"Todt","sequence":"first","affiliation":[{"name":"Department of Molecular and Medical Virology, Ruhr University Bochum, 44801 Bochum, Germany;"},{"name":"Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany;"},{"name":"European Virus Bioinformatics Center (EVBC), 07743 Jena, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Friesland","sequence":"additional","affiliation":[{"name":"Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nora","family":"Moeller","sequence":"additional","affiliation":[{"name":"Department of Molecular and Medical Virology, Ruhr University Bochum, 44801 Bochum, Germany;"},{"name":"Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint 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School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yannick","family":"Br\u00fcggemann","sequence":"additional","affiliation":[{"name":"Department of Molecular and Medical Virology, Ruhr University Bochum, 44801 Bochum, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonard","family":"Knegendorf","sequence":"additional","affiliation":[{"name":"Department of Molecular and Medical Virology, Ruhr University Bochum, 44801 Bochum, Germany;"},{"name":"Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Burkard","sequence":"additional","affiliation":[{"name":"Department of Molecular and Medical Virology, Ruhr University Bochum, 44801 Bochum, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joerg","family":"Steinmann","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, University Hospital of Essen, 45147 Essen, Germany;"},{"name":"Institute of Clinical Hygiene, Medical Microbiology and Infection, Paracelsus Medical University, 90419 N\u00fcrnberg, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rani","family":"Burm","sequence":"additional","affiliation":[{"name":"Laboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, Belgium;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lieven","family":"Verhoye","sequence":"additional","affiliation":[{"name":"Laboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 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R.","family":"Vondran","sequence":"additional","affiliation":[{"name":"Regenerative Medicine and Experimental Surgery (ReMediES), Department of General, Visceral and Transplantation Surgery, Hannover Medical School, 30625 Hannover, Germany;"},{"name":"German Centre for Infection Research (DZIF), Hannover-Braunschweig, 30625 Hannover, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang","family":"Baumg\u00e4rtner","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip","family":"Meuleman","sequence":"additional","affiliation":[{"name":"Laboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, Belgium;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Behrendt","sequence":"additional","affiliation":[{"name":"Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany;"},{"name":"German Centre for Infection Research (DZIF), Hannover-Braunschweig, 30625 Hannover, Germany;"},{"name":"Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eike","family":"Steinmann","sequence":"additional","affiliation":[{"name":"Department of Molecular and Medical Virology, Ruhr University Bochum, 44801 Bochum, Germany;"},{"name":"Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2020,1,2]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"crossref","first-page":"1388","DOI":"10.1053\/j.gastro.2012.02.014","article-title":"Pathogenesis and treatment of hepatitis e virus infection","volume":"142","author":"Wedemeyer H.","year":"2012","unstructured":"H. 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