Integrative Effect of Defective Interfering RNA Accumulation and Helper Virus Attenuation Is Responsible for the Persistent Infection of Japanese Encephalitis Virus in BHK-21 Cells
Integrative Effect of Defective Interfering RNA Accumulation and Helper Virus Attenuation Is Responsible for the Persistent Infection of Japanese Encephalitis Virus in BHK-21 Cells
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DOI：
10.1002/jmv.23665
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发表时间：
2013-11-01
影响因子：
12.7
通讯作者：
Jeong, Yong Seok
中科院分区：
文献类型：
--
作者：
Park, Soo Young;Choi, Eunmi;Jeong, Yong Seok
RNA 病毒的持续存在通常（但并非总是）与有缺陷的干扰 (DI) 颗粒的产生有关。为了研究 DI 颗粒和辅助病毒在 RNA 病毒持久性中的可能作用，在幼仓鼠肾 (BHK-21) 细胞中建立了日本脑炎病毒 (JEV) 的持续感染。 JEV 在 BHK-21 细胞上连续第 6 次和第 7 次未稀释传代时，随着 DI RNA 的生成，病毒的持久性自发建立。从表现出 JEV 持久性的最初 BHK-21 细胞批次中获得了 7 个表现出持续感染的细胞克隆，并维持了 400 多天。大多数细胞克隆持续产生感染性颗粒(10(1)-10(5)PFU/ml)，表达病毒蛋白，并抵抗同源重复感染。从七个细胞克隆中的两个分离出两种辅助病毒 chvBS6-3 和 chvBS7-1，并对其进行表征以研究它们在 JEV 持久性中的作用。虽然 chvBS6-3 在没有 DI RNA 的情况下恢复到其完全细胞病变性，但 chvBS7-1 几乎没有表现出细胞病变性，无论 DI RNA 共复制如何。 chvBS7-1 的减毒似乎并不是由于吸附或基因组复制不充分，而是由于组装的子代病毒粒子的低效流出，这表明 JEV 在 BHK-21 细胞中持续存在期间辅助病毒的出现发生了改变。这些观察结果表明，JEV 的持久性至少涉及两种机制： DI RNA 依赖性机制，其中 DI RNA 共复制使辅助病毒的细胞病变性无效，或 DI RNA 独立机制，其中辅助病毒自我减毒。这项研究为了解 RNA 病毒持续感染的机制提供了一个有用的体外工具。医学杂志。病毒。 85：1990-2000，2013 年。(c) 2013 年 Wiley 期刊公司。
Persistence of RNA viruses is often, but not always, associated with the production of defective interfering (DI) particles. To investigate possible roles of DI particles and helper viruses in RNA virus persistence, persistent infection with Japanese encephalitis virus (JEV) was established in baby hamster kidney (BHK-21) cells. At the 6th and 7th serial undiluted passages of JEV on BHK-21 cells, viral persistence was established spontaneously with DI RNA generation. Seven cell clones exhibiting persistent infection were obtained from the initial BHK-21 cell batches exhibiting JEV persistence, and maintained for over 400 days. Most cell clones produced infectious particles (10(1)-10(5)PFU/ml) continuously, expressed viral proteins, and resisted homologous superinfection. Two helper viruses, chvBS6-3 and chvBS7-1, were isolated from two of the seven cell clones, and characterized to investigate their roles in JEV persistence. While chvBS6-3 was restored to its full cytopathicity in the absence of DI RNA, chvBS7-1 exhibited almost no cytopathicity, regardless of DI RNA co-replication. Attenuation of chvBS7-1 did not appear to be due to inadequate adsorption or genome replication, but due to inefficient egress of the assembled progeny virions, suggesting altered helper virus emergence during JEV persistence in BHK-21 cells. These observations suggest that at least two mechanisms are involved in JEV persistence; a DI RNA-dependent mechanism, where DI RNA co-replication nullifies the helper virus's cytopathicity, or a DI RNA-independent mechanism, where the helper virus is self-attenuated. This study provides a useful in vitro tool for understanding the mechanisms underlying RNA virus persistent infections. J Med. Virol. 85:1990-2000, 2013. (c) 2013 Wiley Periodicals, Inc.