Schemmerer Mathias, Apelt Silke, Trojnar Eva, Ulrich Rainer G, Wenzel Jürgen J, Johne Reimar
Institute of Clinical Microbiology and Hygiene, 93053 Regensburg, Germany.
German Federal Institute for Risk Assessment, Department of Biological Safety, 12277 Berlin, Germany.
Viruses. 2016 Sep 29;8(10):267. doi: 10.3390/v8100267.
Hepatitis E virus (HEV) is a human pathogen with increasing importance. The lack of efficient cell culture systems hampers systematic studies on its replication cycle, virus neutralization and inactivation. Here, several cell lines were inoculated with the HEV genotype 3c strain 47832c, previously isolated from a chronically infected transplant patient. At 14 days after inoculation the highest HEV genome copy numbers were found in A549 cells, followed by PLC/PRF/5 cells, whereas HepG2/C3A, Huh-7 Lunet BLR and MRC-5 cells only weakly supported virus replication. Inoculation of A549-derived subclone cell lines resulted in most cases in reduced HEV replication. However, the subclone A549/D3 was susceptible to lower virus concentrations and resulted in higher virus yields as compared to parental A549 cells. Transcriptome analysis indicated a downregulation of genes for () and , and an upregulation of the () gene in A549/D3 cells compared to A549 cells. However, treatment of A549/D3 cells or A549 cells with CEACAM- or syndecan 2-specific antisera did not influence HEV replication. The results show that cells supporting more efficient HEV replication can be selected from the A549 cell line. The specific mechanisms responsible for the enhanced replication remain unknown.
戊型肝炎病毒（HEV）是一种愈发重要的人类病原体。缺乏高效的细胞培养系统阻碍了对其复制周期、病毒中和及灭活的系统性研究。在此，用先前从一名慢性感染的移植患者中分离出的戊型肝炎病毒3c基因型菌株47832c接种了几种细胞系。接种后14天，在A549细胞中发现了最高的戊型肝炎病毒基因组拷贝数，其次是PLC/PRF/5细胞，而HepG2/C3A、Huh-7 Lunet BLR和MRC-5细胞仅微弱支持病毒复制。接种A549衍生的亚克隆细胞系在大多数情况下导致戊型肝炎病毒复制减少。然而，与亲代A549细胞相比，亚克隆A549/D3对较低的病毒浓度敏感，并产生了更高的病毒产量。转录组分析表明，与A549细胞相比，A549/D3细胞中（）和的基因下调，而（）基因上调。然而，用CEACAM或syndecan 2特异性抗血清处理A549/D3细胞或A549细胞并不影响戊型肝炎病毒复制。结果表明，可以从A549细胞系中选择支持更高效戊型肝炎病毒复制的细胞。导致复制增强的具体机制仍不清楚。
