Brown Hannah M, Marlet Julien, León-Janampa Nancy, Brand Denys, Fletcher Nicola F
Veterinary Sciences Centre and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
INSERM U1259 MAVIVHe, CHRU de Tours, Université de Tours et CHRU de Tours and Service de Bactériologie-Virologie-Hygiène, Tours, France.
Sci Rep. 2025 Mar 4;15(1):7598. doi: 10.1038/s41598-025-92164-9.
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.
戊型肝炎病毒（HEV）是全球急性病毒性肝炎的主要病因，也是唯一的人畜共患型肝炎病毒。HEV 基因型 3（HEV3）与一系列临床表现有关，包括发达国家免疫功能低下个体的慢性感染以及欧洲偶发性的本地 HEV3 病例。目前的体外模型支持的 HEV 感染水平较低，这妨碍了我们对病毒发病机制的理解以及治疗方法的开发。我们为两种广泛使用的肝癌细胞系 PLC-PRF-5 和 Huh-7.5 开发了改良培养方法，并评估了 HEV 感染情况。PLC-PRF-5 细胞形成了简单的上皮样极性和分化，紧密连接蛋白闭合蛋白和闭合蛋白 1 定位于细胞间连接以及白蛋白产量增加证明了这一点。在 Huh-7.5 细胞中观察到了复杂的肝细胞样极性，紧密连接蛋白定位于共享的内部胆小管样结构，并且荧光分子 5(6)-羧基荧光素二乙酸酯得以保留。用基因型 3 HEV 感染细胞，通过 RT-qPCR 以及对 HEV ORF2 和双链 RNA 进行免疫荧光标记，观察到 HEV 的感染和复制增强。我们描述了用于体外 HEV 感染的强大且易于操作的模型。这些模型将有助于开展研究，加深我们对这种新出现的人畜共患病原体的理解，并开发治疗干预措施。
