Medical Faculty, Kunming University of Science and Technology, Kunming, PR China.
Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming, PR China.
J Med Virol. 2021 Nov;93(11):6267-6277. doi: 10.1002/jmv.27121. Epub 2021 Jun 10.
Hepatitis E virus (HEV) is the major pathogen of viral hepatitis. However, the understanding of the HEV life cycle is limited. In the present study, cells were separately infected with nonenveloped HEV (derived from feces or bile) or quasi-enveloped HEV (derived from the cell culture after serial passages, eHEV) and observed by confocal fluorescence microscopy to investigate the life cycle of HEV. HEV finished its binding and entry into host cells at first 6 h postinoculation (hpi). Cells inoculated with eHEV showed less infectivity than cells inoculated with nonenveloped HEV. Newly synthesized progeny virions were released into the supernatant of cell cultures from 48 hpi. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis results showed that the supernatant's progeny viruses were infectious even after five serial passages. These results show the significant difference between nonenveloped HEV and eHEV, which will provide novel insights into the HEV replication cycle. The efficient cell culture of HEV will promote the development of anti-HEV drugs and vaccines.
戊型肝炎病毒（HEV）是病毒性肝炎的主要病原体。然而，人们对 HEV 生命周期的认识有限。在本研究中，分别用无包膜 HEV（源自粪便或胆汁）或准包膜 HEV（源自连续传代后的细胞培养物，eHEV）感染细胞，并通过共聚焦荧光显微镜观察，以研究 HEV 的生命周期。HEV 在接种后 6 小时（hpi）内完成结合和进入宿主细胞的过程。用 eHEV 接种的细胞比用无包膜 HEV 接种的细胞的感染性低。从 48 hpi 开始，新合成的子代病毒颗粒被释放到细胞培养物的上清液中。实时荧光定量聚合酶链反应（qRT-PCR）和 Western blot 分析结果表明，即使经过五次连续传代，上清液中的子代病毒仍具有感染性。这些结果表明无包膜 HEV 和 eHEV 之间存在显著差异，这将为 HEV 复制周期提供新的见解。HEV 的高效细胞培养将促进抗 HEV 药物和疫苗的开发。
