Event & Pasteur Colloquium
[2019 Pasteur Colloquium] Dr. Daniel Todt, Ruhr University Bochum (Germany)
October 08, 2019
IPK Pasteur Colloquium
Date : Thur. Oct. 10, 2019 at 11:00am
Venue : Monod Room (1F), Institut Pasteur Korea
Daniel Todt, Ph.D.
Post-Doc, Department of Molecular and Medical Virology,
Ruhr University Bochum, Germany
Robust hepatitis E virus infection and transcriptional response in human hepatocytes
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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 Orthohepevirus within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle and development of effective antivirals and vaccines 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 HEV cell culture-derived particles (HEVcc) with viral titers between 105 to 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 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 the off-label drug 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 and vaccines for this important zoonotic pathogen.
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