In vitro inhibition of fish rhabdoviruses by Japanese flounder, Paralichthys olivaceus Mx
In vitro inhibition of fish rhabdoviruses by Japanese flounder, Paralichthys olivaceus Mx
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DOI：
10.1016/j.virol.2003.08.040
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发表时间：
2003-12-20
期刊：
影响因子：
3.7
通讯作者：
Aoki, T
中科院分区：
文献类型：
--
作者：
Caipang, CMA;Hirono, I;Aoki, T
用一种稳定表达重组牙鲆Mx （JFMx）的同源鱼细胞系感染了横纹肌病毒（hirame rhabdovirus， HIRRV）和病毒性出血性败血症病毒（viral hemorrhagic sepsis virus， VHSV），这两种病毒均为横纹肌病毒科阴性单链RNA病毒。两种横纹肌病毒的初代转录分析表明，转染jfmx的细胞系中病毒核蛋白转录物的表达水平和拷贝数低于感染对照细胞，但差异不显著。这表明JFMx可能不是横纹肌病毒初级转录的有效抑制剂。RT-PCR和实时定量RT-PCR检测的横纹肌病毒表达动力学显示，随着时间的推移，横纹肌病毒糖蛋白和核蛋白转录物水平降低，这表明JFMx可能通过干扰病毒亚基因组mrna的转录来阻断横纹肌病毒的复制。对横纹肌病毒复制的显著抑制导致病毒颗粒的合成减少。这也许可以解释为什么表达jfmx的细胞对病毒诱导的细胞裂解不那么敏感，因此，为什么它们的存活率明显高于受感染的对照细胞。这些结果为JFMx在体外具有抗病毒作用提供了直接证据。(C) 2003 Elsevier Inc.版权所有。
A homologous fish cell line stably expressing the recombinant Japanese flounder Mx (JFMx) was infected with hirame rhabdovirus (HIRRV) and viral hemorrhagic septicemia virus (VHSV), both of which are negative single-stranded RNA viruses belonging to the Rhabdoviridae family. Analysis of primary transcription of the two rhabdoviruses showed that there was lower expression level and copy number of the viral nucleoprotein transcript in the JFMx-transfected cell line than the infected, control cells, although no significant difference was observed. This suggests that JFMx may not be a potent inhibitor of rhabdoviral primary transcription. Kinetics of rhabdovirus expression by RT-PCR and quantitative real-time RT-PCR showed reduced levels of the rhabdoviral glycoprotein and nucleoprotein transcripts over time, indicating the possible role of JFMx in blocking rhabdoviral replication by interfering with the transcription of the viral subgenomic mRNAs. Significant inhibition in rhabdovirus replication consequently resulted in the synthesis of fewer viral particles. This may explain why JFMx-expressing cells are less susceptible to virus-induced cell lysis, and thus, why they would have a significantly higher survival than the infected, control cells. These results provide direct evidence that JFMx has an antiviral effect in vitro. (C) 2003 Elsevier Inc. All rights reserved.