Laboratory of Molecular Virology and Biotechnology, Institute of Biotechnology, National Cheng-Kung University, Tainan 701, Taiwan.
Department of Life Sciences, National University of Kaohsiung, Kaohsiung 811, Taiwan.
Virus Res. 2016 Feb 2;213:37-45. doi: 10.1016/j.virusres.2015.11.003. Epub 2015 Nov 10.
Giant seaperch iridovirus (GSIV) induces cell death by an unknown mechanism. We postulated that this mechanism involves mitochondria-mediated cell death. Cell viability assays revealed a steady increase in dead grouper fin cells (GF-1) after GSIV infection, from 11% at 2 days post-infection (dpi) to 67% at 5 dpi. Annexin V/PI staining revealed GSIV infection induced apoptosis in a steadily increasing fraction of cells, from 4% at 1 dpi to 29% at 5 dpi. Furthermore, post-apoptotic necrosis was apparent at 4 and 5 dpi in the late replication stage. In the early replication stage, JC-1 dye revealed mitochondrial membrane potential (ΔΨm) loss in 42% of infected cells at 1 dpi, increasing to 98% at 3 dpi. Phosphatidylserine (PS) exposure and loss of ΔΨm from apoptosis/necrosis was attenuated by treatment with the adenine nucleotide translocase inhibitor bongkrekic acid (BKA) and the protein synthesis inhibitor cyclohexamide (CHX). These data suggest GSIV induces GF-1 apoptotic/necrotic cell death through pathways that require newly synthesized protein and involve the mitochondrial function.
巨型石斑鱼虹彩病毒（GSIV）通过未知机制诱导细胞死亡。我们推测这种机制涉及线粒体介导的细胞死亡。细胞活力测定显示，GSIV 感染后，石斑鱼鳍细胞（GF-1）的死亡细胞数量持续增加，从感染后 2 天的 11%增加到第 5 天的 67%。Annexin V/PI 染色显示，GSIV 感染以稳定增加的比例诱导细胞凋亡，从感染后 1 天的 4%增加到第 5 天的 29%。此外，在晚期复制阶段的第 4 和第 5 天，可见凋亡后坏死。在早期复制阶段，JC-1 染料显示，在第 1 天感染的 42%细胞中，线粒体膜电位（ΔΨm）丧失，在第 3 天增加到 98%。用腺嘌呤核苷酸转位酶抑制剂绷酸（BKA）和蛋白质合成抑制剂环己酰胺（CHX）处理可减轻凋亡/坏死时 PS 暴露和 ΔΨm 的丧失。这些数据表明，GSIV 通过需要新合成蛋白质并涉及线粒体功能的途径诱导 GF-1 细胞发生凋亡/坏死性死亡。
