Characterization of a Cell Culture System of Persistent Hepatitis E Virus Infection in the Human HepaRG Hepatic Cell Line

Abstract

Hepatitis E virus (HEV) is considered as an emerging global health problem. In most cases, hepatitis E is a self-limiting disease and the virus is cleared spontaneously without the need of antiviral therapy. However, immunocompromised individuals can develop chronic infection and liver fibrosis that can progress rapidly to cirrhosis and liver failure. The lack of efficient and relevant cell culture system and animal models has limited our understanding of the biology of HEV and the development of effective drugs for chronic cases. In the present study, we developed a model of persistent HEV infection in human hepatocytes in which HEV replicates efficiently. This HEV cell culture system is based on differentiated HepaRG cells infected with an isolate of HEV-3 derived from a patient suffering from acute hepatitis E. Efficient replication was maintained for several weeks to several months as well as after seven successive passages on HepaRG naïve cells. Moreover, after six passages onto HepaRG, we found that the virus was still infectious after oral inoculation into pigs. We also showed that ribavirin had an inhibitory effect on HEV replication in HepaRG. In conclusion, this system represents a relevant and efficient in vitro model of HEV replication that could be useful to study HEV biology and identify effective antiviral drugs against chronic HEV infection.

Modeling Hepatotropic Viral Infections: Cells vs. Animals

“…They reported that these in vitro culture systems support HEV replication and release of encapsulated RNA [43]. In accordance with this study, Pellerin and her colleagues showed that HepaRG is a relevant and efficient in vitro model of HEV replication that could be used to study HEV and identify effective antiviral drugs against chronic HEV infection [44]. Besides, another study introduced a new procedure using a cocktail of 5 chemicals (Forskolin, SB431542, IWP2, DAPT, and LDN193189), allowing fast differentiation and efficient HDV-infection of HepaRG cells [45].…”

“…Primary cells Adult PHH [15,21] Adult PHH [17] Adult PHH [18][19][20][21] Adult PHH [18] primary human intestinal cells [23] primary human endometrial cells [24] Cancer cell lines HepAD38 [34] HepDE19 [35] Hepa RG [44,46] HLCZ01 [36] (HepNB2.7) [38] HrpaRG [39,40] HepG2 +Huh-7 [49,55] HepG2 [51] HepG2-NTCP [52] HLC [24,26,64] HLCZ01 [36] HepG2 [ [150] Abbreviations: ETV, entecavir; FRG, fumaryl acetoacetate hydrolase (Fah)/RAG2/interleukin (IL) 2-gammaC; FRGS, Fah −/− Rag2 −/− IL-2Rγc −/− SCID; HEV, hepatitis E virus; HBV, hepatitis B virus; HCV, hepatitis C virus; IFN, interferon; IG, immunoglobulin; NK, natural killer; NM23TC, nanoformulation of the modified lamivudine 3TC; PRI-724, selective inhibitor of β-catenin/CBP; RG7834, inhibitor of HBV expression; uPA/RAG-2, urokinase-type plasminogen activator/recombinant activation gene-2; uPA/SCID, urokinase-type plasminogen activator/severe combined immunodeficiency; WHBV, woodchuck hepatitis B virus; WMHBV, woolly monkey hepatitis B virus.…”

Investigations on SARS-CoV-2 and other coronaviruses in mink farms in France at the end of the first year of COVID-19 pandemic

“…The RNA library was prepared for each RNA sample tested using Ion Total RNA-Seq kit v2 (Life Technologies, Carlsbad, CA, USA) and then sequenced using Ion Torrent Proton technology. The reads were cleaned with the Trimmomatic 0.36 software, followed by bioinformatics analysis as previously described (37,38) using the GenBank Mink Caliciviridae reference sequence MF677852.1 for the two swab RNA samples and the Mink coronavirus 1 reference sequence MN535737.1 for the fecal RNA sample to calculate sub-sampling and final alignment.…”

Low prevalence of hepatitis E virus in the liver of Corsican pigs slaughtered after 12 months despite high antibody seroprevalence

“…HepaRG cell culture: Human HepaRG™ cells were grown as previously described (Pellerin et al., 2021). Briefly, HepaRG were seeded into 6‐well plates (2 × 10 5 cells/well) and maintained in growth medium for 2 weeks.…”