Development of an in vitro system to measure the sensitivity to the antiviral Mx protein of fish viruses
Development of an in vitro system to measure the sensitivity to the antiviral Mx protein of fish viruses
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DOI：
10.1016/j.jviromet.2012.01.014
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发表时间：
2012-06-01
影响因子：
3.1
通讯作者：
Collet, Bertrand
中科院分区：
文献类型：
--
作者：
Lester, Katherine;Hall, Malcolm;Collet, Bertrand
Mx是一种结构蛋白，由I型干扰素（IFN）诱导，具有直接的抗病毒特性。在鱼类中，Mx蛋白对病毒保护的固有贡献是未知的。对转基因奇努克鲑鱼胚胎（CHSE）-TOF细胞系进行遗传修饰，使其在四环素衍生物强力霉素（DOX）的控制下表达虹鳟Mx（rbtMx 1）蛋白。分离两个克隆CHSE-TOF-MX 8和CHSE-TOF-MX 10并通过qPCR表征。测量在有和没有DOX的情况下培养的CHSE-TOF、CHSE-TOF-MX 8和CHSE-TOF-MX 10细胞系对传染性胰腺坏死病毒（IPNV）、鲑鱼甲病毒（SAV）、传染性造血坏死病毒（IHNV）和流行性造血坏死病毒（EHNV）的抗性水平。建立了一种新的方法，以准确地测量任何给定的病毒分离物对Mx蛋白的敏感性水平。IPNV和SAV病毒对细胞中存在的rbtMx 1高度敏感，而IHNV和EHNV显示出部分抗性，这表明这些类别的病毒之间的对比病毒逃避策略。皇冠版权所有（C）2012由Elsevier B. V.出版。保留所有权利。
Mx is a structural protein, induced by type I interferon (IFN), with direct antiviral properties. In fish the inherent contribution of Mx protein to viral protection is unknown. The transgenic Chinook salmon embryonic (CHSE)-TOF cell line was genetically modified to express the rainbow trout Mx (rbtMx1) protein under the control of the tetracycline derivative, doxycycline (DOX). Two clones CHSE-TOF-MX8 and CHSE-TOF-MX10 were isolated and characterised by qPCR. The level of resistance to Infectious Pancreatic Necrosis Virus (IPNV), Salmon Alphavirus (SAV), Infectious Haematopoietic Necrosis Virus (IHNV) and Epizootic Haematopoietic Necrosis Virus (EHNV) of the CHSE-TOF, CHSE-TOF-MX8 and CHSE-TOF-MX10 cell lines cultivated with and without DOX was measured. A novel method was established to measure accurately the level of sensitivity of any given viral isolate to Mx protein. IPNV and SAV viruses were highly sensitive to the presence of rbtMx1 in the cells whereas IHNV and EHNV showed partial resistance suggesting contrasting viral evasion strategies between these categories of viruses. Crown Copyright (C) 2012 Published by Elsevier B.V. All rights reserved.