Characterization of two grouper Epinephelus akaara cell lines: Application to studies of Singapore grouper iridovirus (SGIV) propagation and virus-host interaction
Characterization of two grouper Epinephelus akaara cell lines: Application to studies of Singapore grouper iridovirus (SGIV) propagation and virus-host interaction
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DOI：
10.1016/j.aquaculture.2009.04.019
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发表时间：
2009-07-15
期刊：
影响因子：
4.5
通讯作者：
Qin, Qiwei
中科院分区：
文献类型：
--
作者：
Huang, Xiaohong;Huang, Youhua;Qin, Qiwei
建立连续的鱼类细胞系对于研究病毒病理学和宿主-病毒相互作用是必不可少的。本研究以赤点石斑鱼（Epinealopus akaara）的脾脏和鱼鳔为材料，建立了两个细胞系，命名为EAGS和EAGSB。在25 - 30 ℃的温度下，两种细胞在补充有10%胎牛血清的Leibovitz's L-15培养基中增殖良好。染色体分析表明，EAGS和EAGSB的众数分别为88和72。两种细胞系均对新加坡石斑鱼虹彩病毒（SGIV）敏感，如细胞病变效应（CPE）和病毒滴度增加所示。电子显微镜和免疫荧光显微镜进一步证实了SGIV在EAGS和EAGSB细胞中的复制。然而，这两种细胞系对鳖虹彩病毒（STIV）、病毒性神经坏死病毒（VNNV）和鲤鱼春季病毒血症病毒（SVCV）均不敏感。Hoechst染色显示SGIV感染的EAGS细胞中未检测到凋亡小体，提示SGIV感染诱导的EAGS细胞死亡可能与典型的凋亡不同。此外，SGIV感染在EAGS中改变了微管的形态和结构，包括重排成围绕核的环状结构，以及围绕病毒组装位点的聚集。所有这些数据表明，EAGS和EAGSB细胞具有良好的潜力，研究SGIV和宿主细胞之间的相互作用。(C)2009爱思唯尔有限公司版权所有。
The establishment of continuous fish cell lines is essential for studying viral pathology and host-virus interactions. In the present study, two cell lines, designated as EAGS and EAGSB, were established from the spleen and swim bladder of grouper, Epinephelus akaara. Both cells multiplied well in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum, at temperatures between 25 and 30 degrees C. Chromosome analysis revealed that modal chromosome number was 88 and 72 for EAGS and EAGSB, respectively. Both cell lines were susceptible to Singapore grouper iridovirus (SGIV), as shown by cytopathic effect (CPE) and increased viral titers. SGIV replication in EAGS and EAGSB cells was further confirmed by electron microscopy and immunofluorescence microscopy. However, both cell lines were not susceptible to soft-shelled turtle iridovirus (STIV), viral nervous necrosis virus (VNNV) and spring viremia of carp virus (SVCV). Hoechst staining showed that no apoptotic bodies were examined in SGIV infected cells, suggesting that SGIV infection induced EAGS cells death might be distinguished from typical apoptosis. Moreover, SGIV infection in EAGS altered the morphology and structure of microtubules, including rearrangement into a ring-like structure around the nucleus, and aggregations around the virus assembly sites. All these data imply that EAGS and EAGSB cells have good potential for investigation of the interactions between SGIV and host cells. (C) 2009 Elsevier B.V. All rights reserved.