title: Hepatitis E Virus (HEV) Genotype 3 Infection of Human Liver Chimeric Mice as a Model for Chronic HEV Infection.
abstract: Genotype 3 (gt3) hepatitis E virus (HEV) infections are emerging in Western countries. Immunosuppressed patients are at risk of chronic HEV infection and progressive liver damage, but no adequate model system currently mimics this disease course. Here we explore the possibilities of in vivo HEV studies in a human liver chimeric mouse model (uPA(+/+)Nod-SCID-IL2Rγ(-/-)) next to the A549 cell culture system, using HEV RNA-positive EDTA-plasma, feces, or liver biopsy specimens from 8 immunocompromised patients with chronic gt3 HEV. HEV from feces- or liver-derived inocula showed clear virus propagation within 2 weeks after inoculation onto A549 cells, compared to slow or no HEV propagation of HEV RNA-positive, EDTA-plasma samples. These in vitro HEV infectivity differences were mirrored in human-liver chimeric mice after intravenous (i.v.) inoculation of selected samples. HEV RNA levels of up to 8 log IU HEV RNA/gram were consistently present in 100% of chimeric mouse livers from week 2 to week 14 after inoculation with human feces- or liver-derived HEV. Feces and bile of infected mice contained moderate to large amounts of HEV RNA, while HEV viremia was low and inconsistently detected. Mouse-passaged HEV could subsequently be propagated for up to 100 days in vitro In contrast, cell culture-derived or seronegative EDTA-plasma-derived HEV was not infectious in inoculated animals. In conclusion, the infectivity of feces-derived human HEV is higher than that of EDTA-plasma-derived HEV both in vitro and in vivo Persistent HEV gt3 infections in chimeric mice show preferential viral shedding toward mouse bile and feces, paralleling the course of infection in humans.
  
  
  OBJECTIVE
  Hepatitis E virus (HEV) genotype 3 infections are emerging in Western countries and are of great concern for immunosuppressed patients at risk for developing chronic HEV infection. Lack of adequate model systems for chronic HEV infection hampers studies on HEV infectivity and transmission and antiviral drugs. We compared the in vivo infectivity of clinical samples from chronic HEV patients in human liver chimeric mice to an in vitro virus culture system. Efficient in vivo HEV infection is observed after inoculation with feces- and liver-derived HEV but not with HEV RNA-containing plasma or cell culture supernatant. HEV in chimeric mice is preferentially shed toward bile and feces, mimicking the HEV infection course in humans. The observed in vivo infectivity differences may be relevant for the epidemiology of HEV in humans. This novel small-animal model therefore offers new avenues to unravel HEV's pathobiology.
authors.affiliation: Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Martijn D B
authors.identifier: http://orcid.org/0000-0003-0509-7713
authors.initials: MD
authors.lastname: van de Garde
authors.affiliation: Department of Viroscience, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Suzan D
authors.initials: SD
authors.lastname: Pas
authors.affiliation: Department of Viroscience, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Guido
authors.initials: G
authors.lastname: van der Net
authors.affiliation: Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Robert A
authors.initials: RA
authors.lastname: de Man
authors.affiliation: Department of Viroscience, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Albert D M E
authors.initials: AD
authors.lastname: Osterhaus
authors.affiliation: Department of Viroscience, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Bart L
authors.initials: BL
authors.lastname: Haagmans
authors.affiliation: Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, The Netherlands.
authors.forename: Andre
authors.initials: A
authors.lastname: Boonstra
authors.affiliation: Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, The Netherlands Department of Gastroenterology and Hepatology, University Hospital Antwerp, Antwerp, Belgium t.vanwolleghem@erasmusmc.nl.
authors.forename: Thomas
authors.initials: T
authors.lastname: Vanwolleghem
mesh_terms: D000818:Animals; D002460:Cell Line; D002908:Chronic Disease; D004195:Disease Models, Animal; D005838:Genotype; D016751:Hepatitis E / Q000821:virology*; D016752:Hepatitis E virus / Q000235:genetics* / Q000378:metabolism; D022781:Hepatocytes / Q000637:transplantation / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D051379:Mice; D019562:Viral Load; D014779:Virus Replication