Rat hepatitis E virus derived from wild rats (Rattus rattus) propagates efficiently in human hepatoma cell lines
Rat hepatitis E virus derived from wild rats (Rattus rattus) propagates efficiently in human hepatoma cell lines
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DOI：
10.1016/j.virusres.2014.03.002
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发表时间：
2014-06-24
期刊：
影响因子：
5
通讯作者：
Okamoto, Hiroaki
中科院分区：
文献类型：
--
作者：
Jirintai, Suljid;Tanggis;Okamoto, Hiroaki
尽管已在野生大鼠中发现了大鼠戊型肝炎病毒（HEV），但尚未建立该病毒的细胞培养系统。最近的一份报告表明，人类血清中存在针对大鼠 HEV 的抗体，这鼓励我们在人类细胞中培养大鼠 HEV。当从印度尼西亚野生大鼠（Rattusrattus）获得的肝匀浆接种到人肝癌细胞上时，大鼠HEV在PLC/PRF/5、HuH-7和HepG2细胞中有效复制，无论其遗传组（G1-G3）如何。从培养细胞中释放的大鼠 HEV 颗粒表面含有脂质相关膜，这些膜经过去污剂和蛋白酶处理后已被耗尽，蔗糖中的浮力密度从 1.15-1.16 g/ml 变为 1.27-1.28 g/ml。 Northern 印迹分析显示受感染细胞中存在 7.0 kb 的基因组 RNA 和 2.0 kb 的亚基因组 RNA。 G1-G3的亚基因组RNA各自具有GUAGC（nt 4933-4937）的极端5'端序列，位于高度保守的GAAUAACA序列（nt 4916-4923）的下游。大鼠 HEV 培养系统的建立将有助于对病毒复制机制和 HEV 蛋白的功能作用进行深入研究。需要进一步研究以澄清大鼠 HEV 的人畜共患潜力。 (C) 2014 Elsevier B.V. 保留所有权利。
Although rat hepatitis E virus (HEV) has been identified in wild rats, no cell culture systems for this virus have been established. A recent report suggesting the presence of antibodies against rat HEV in human sera encouraged us to cultivate rat HEV in human cells. When liver homogenates obtained from wild rats (Rattus rattus) in Indonesia were inoculated onto human hepatocarcinoma cells, the rat HEV replicated efficiently in PLC/PRF/5, HuH-7 and HepG2 cells, irrespective of its genetic group (G1-G3). The rat HEV particles released from cultured cells harbored lipid-associated membranes on their surface that were depleted by treatment with detergent and protease, with the buoyant density in sucrose shifting from 1.15-1.16 g/ml to 1.27-1.28 g/ml. A Northern blotting analysis revealed genomic RNA of 7.0 kb and subgenomic RNA of 2.0 kb in the infected cells. The subgenomic RNA of G1-G3 each possessed the extreme 5'-end sequence of GUAGC (nt 4933-4937), downstream of the highly conserved sequence of GAAUAACA (nt 4916-4923). The establishment of culture systems for rat HEV would allow for extended studies of the mechanisms of viral replication and functional roles of HEV proteins. Further investigation is required to clarify the zoonotic potential of rat HEV. (C) 2014 Elsevier B.V. All rights reserved.