National Consultant Laboratory for HAV and HEV, Institute of Clinical Microbiology and Hygiene, University Medical Center Regensburg, 93053 Regensburg, Germany.
Viruses. 2022 May 23;14(5):1116. doi: 10.3390/v14051116.
The family comprises the species (HEV-A to -D). HEV-C genotype 1 (HEV-C1, rat HEV) is able to infect humans. This study investigated whether an optimized HEV-A cell culture system is able to propagate the cell culture-derived rat HEV, and if de novo isolation of the virus from rat liver is possible. We tested the liver carcinoma cell lines PLC/PRF/5, HuH-7, and HuH-7-Lunet BLR for their susceptibility to HEV-C1 strains. Cells were infected with the cell culture-derived HEV-C1 strain R63 and rat liver-derived strain R68. Cells were maintained in MEMM medium, which was refreshed every 3-4 days. The viral load of HEV-C1 was determined by RT-qPCR in the supernatant and expressed as genome copies per mL (c/mL). Rat HEV replication was most efficient in the newly introduced HuH-7-Lunet BLR cell line. Even if the rat HEV isolate had been pre-adapted to PLC/PRF/5 by multiple passages, replication in HuH-7-Lunet BLR was still at least equally effective. Only HuH-7-Lunet BLR cells were susceptible to the isolation of HEV-C1 from the liver homogenate. These results suggest HuH-7-Lunet BLR as the most permissive cell line for rat HEV. Our HEV-C1 cell culture system may be useful for basic research, the animal-free generation of large amounts of the virus as well as for the testing of antiviral compounds and drugs.
该家族包括物种（HEV-A 至 -D）。HEV-C 基因型 1（HEV-C1，大鼠 HEV）能够感染人类。本研究旨在探讨优化的 HEV-A 细胞培养系统是否能够增殖细胞培养衍生的大鼠 HEV，以及是否有可能从大鼠肝脏中分离出新的病毒。我们测试了肝癌细胞系 PLC/PRF/5、HuH-7 和 HuH-7-Lunet BLR 对 HEV-C1 株的易感性。用细胞培养衍生的 HEV-C1 株 R63 和大鼠肝脏衍生的株 R68 感染细胞。细胞在 MEMM 培养基中培养，每隔 3-4 天更新一次。通过 RT-qPCR 在上清液中测定 HEV-C1 的病毒载量，并表示为每毫升基因组拷贝数（c/mL）。大鼠 HEV 在新引入的 HuH-7-Lunet BLR 细胞系中复制效率最高。即使大鼠 HEV 分离株已通过多次传代预先适应于 PLC/PRF/5，在 HuH-7-Lunet BLR 中的复制效率仍然至少同样有效。只有 HuH-7-Lunet BLR 细胞对从肝匀浆中分离 HEV-C1 敏感。这些结果表明 HuH-7-Lunet BLR 是大鼠 HEV 最允许的细胞系。我们的 HEV-C1 细胞培养系统可能对基础研究、无动物大量生成病毒以及抗病毒化合物和药物的测试有用。
