Enhanced hepatitis E virus infection of polarised hepatocytes in vitro
Abstract Hepatitis E virus (HEV) is a leading cause of acute viral hepatitis worldwide, and the only zoonotic hepatitis virus. HEV genotype 3 (HEV3) is associated with a range of clinical presentations including chronic infection in immunocompromised individuals in developed nations as well as spora...
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Nature Portfolio
2025-03-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-025-92164-9 |
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| author | Hannah M. Brown Julien Marlet Nancy León-Janampa Denys Brand Nicola F. Fletcher |
| author_facet | Hannah M. Brown Julien Marlet Nancy León-Janampa Denys Brand Nicola F. Fletcher |
| author_sort | Hannah M. Brown |
| collection | DOAJ |
| description | Abstract Hepatitis E virus (HEV) is a leading cause of acute viral hepatitis worldwide, and the only zoonotic hepatitis virus. HEV genotype 3 (HEV3) is associated with a range of clinical presentations including chronic infection in immunocompromised individuals in developed nations as well as sporadic cases of autochthonous HEV3 in Europe. Current in vitro models support low levels of HEV infection, hampering our understanding of viral pathogenesis and development of therapeutics. We developed modified culture methods for two widely used hepatoma cell lines, PLC-PRF-5 and Huh-7.5, and evaluated HEV infection. Simple epithelial-like polarity and differentiation formed in PLC-PRF-5 cells, evidenced by localisation of tight junction proteins occludin and zona-occludin 1 to intercellular junctions, and increased albumin production. Complex hepatocyte-like polarity was observed in Huh-7.5 cells, with tight junction proteins localised to shared internal bile canaliculi-like structures and retention of the fluorescent molecule, 5(6)-Carboxyfluorescein diacetate. Cells were infected with genotype 3 HEV, and enhanced infection and replication of HEV was observed using RT-qPCR and immunofluorescent labelling of HEV ORF2 and dsRNA. We describe robust, accessible models for HEV infection in vitro. These models will allow studies to further our understanding of this emerging zoonotic pathogen and develop therapeutic interventions. |
| format | Article |
| id | doaj-art-9361eadc5b6c474aacba67d439cd800e |
| institution | DOAJ |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-9361eadc5b6c474aacba67d439cd800e2025-08-20T03:05:53ZengNature PortfolioScientific Reports2045-23222025-03-0115111610.1038/s41598-025-92164-9Enhanced hepatitis E virus infection of polarised hepatocytes in vitroHannah M. Brown0Julien Marlet1Nancy León-Janampa2Denys Brand3Nicola F. Fletcher4Veterinary Sciences Centre and Conway Institute of Biomolecular and Biomedical Research, University College DublinINSERM U1259 MAVIVHe, CHRU de Tours, Université de Tours et CHRU de Tours and Service de Bactériologie-Virologie-HygièneINSERM U1259 MAVIVHe, Université de Tours et CHRU de ToursINSERM U1259 MAVIVHe, CHRU de Tours, Université de Tours et CHRU de Tours and Service de Bactériologie-Virologie-HygièneVeterinary Sciences Centre and Conway Institute of Biomolecular and Biomedical Research, University College DublinAbstract Hepatitis E virus (HEV) is a leading cause of acute viral hepatitis worldwide, and the only zoonotic hepatitis virus. HEV genotype 3 (HEV3) is associated with a range of clinical presentations including chronic infection in immunocompromised individuals in developed nations as well as sporadic cases of autochthonous HEV3 in Europe. Current in vitro models support low levels of HEV infection, hampering our understanding of viral pathogenesis and development of therapeutics. We developed modified culture methods for two widely used hepatoma cell lines, PLC-PRF-5 and Huh-7.5, and evaluated HEV infection. Simple epithelial-like polarity and differentiation formed in PLC-PRF-5 cells, evidenced by localisation of tight junction proteins occludin and zona-occludin 1 to intercellular junctions, and increased albumin production. Complex hepatocyte-like polarity was observed in Huh-7.5 cells, with tight junction proteins localised to shared internal bile canaliculi-like structures and retention of the fluorescent molecule, 5(6)-Carboxyfluorescein diacetate. Cells were infected with genotype 3 HEV, and enhanced infection and replication of HEV was observed using RT-qPCR and immunofluorescent labelling of HEV ORF2 and dsRNA. We describe robust, accessible models for HEV infection in vitro. These models will allow studies to further our understanding of this emerging zoonotic pathogen and develop therapeutic interventions.https://doi.org/10.1038/s41598-025-92164-9Hepatitis virusHepeviridaeZoonosesIn vitro modelPolarity |
| spellingShingle | Hannah M. Brown Julien Marlet Nancy León-Janampa Denys Brand Nicola F. Fletcher Enhanced hepatitis E virus infection of polarised hepatocytes in vitro Scientific Reports Hepatitis virus Hepeviridae Zoonoses In vitro model Polarity |
| title | Enhanced hepatitis E virus infection of polarised hepatocytes in vitro |
| title_full | Enhanced hepatitis E virus infection of polarised hepatocytes in vitro |
| title_fullStr | Enhanced hepatitis E virus infection of polarised hepatocytes in vitro |
| title_full_unstemmed | Enhanced hepatitis E virus infection of polarised hepatocytes in vitro |
| title_short | Enhanced hepatitis E virus infection of polarised hepatocytes in vitro |
| title_sort | enhanced hepatitis e virus infection of polarised hepatocytes in vitro |
| topic | Hepatitis virus Hepeviridae Zoonoses In vitro model Polarity |
| url | https://doi.org/10.1038/s41598-025-92164-9 |
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