Newcastle disease virus (NDV)-based assay demonstrates interferon-antagonist activity for the NDV V protein and the Nipah virus V, W, and C proteins
Newcastle disease virus (NDV)-based assay demonstrates interferon-antagonist activity for the NDV V protein and the Nipah virus V, W, and C proteins
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DOI：
10.1128/jvi.77.2.1501-1511.2003
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发表时间：
2003-01-01
影响因子：
5.4
通讯作者：
Basler, CF
中科院分区：
文献类型：
--
作者：
Park, MS;Shaw, ML;Basler, CF
我们生成了一种重组新城疫病毒 (NDV)，它在受感染的鸡胚成纤维细胞 (CEF) 中表达绿色荧光蛋白 (GFP)。该病毒对干扰素 (IFN) 敏感，用鸡 α/β 干扰素 (IFN-α/β) 预处理细胞可完全阻断病毒 GFP 表达。预先转染质粒 DNA 可在 CEF 中诱导 IFN 反应并阻断 NDV-GFP 复制。然而，转染已知的 IFN-α/β 系统抑制剂（包括甲型流感病毒 NSI 蛋白和埃博拉病毒 VP35 蛋白）可恢复 NDV-GFP 复制。因此，我们得出结论，NDV-GFP 病毒可用于筛选质粒表达的蛋白质，以确定其抵抗宿主细胞 IFN 反应的能力。使用该系统，我们表明，面对转染诱导的 IFN 反应，NDV V 蛋白或尼帕病毒 V、W 或 C 蛋白的表达可以挽救 NDV-GFP 复制。尼帕病毒（一种对人类高度致命的病原体）的 V 和 W 蛋白也会阻止灵长类动物细胞中 IFN 诱导型启动子的激活。有趣的是，尼帕病毒V蛋白的氨基末端区域与尼帕病毒W的氨基末端相同，足以发挥IFN拮抗剂活性。相比之下，NDV V蛋白的抗IFN活性似乎位于该蛋白的羧基末端区域，该区域与腮腺炎病毒和2型人副流感病毒的V蛋白表现出的IFN拮抗剂活性有关。
We have generated a recombinant Newcastle disease virus (NDV) that expresses the green fluorescence protein (GFP) in infected chicken embryo fibroblasts (CEFs). This virus is interferon (IFN) sensitive, and pretreatment of cells with chicken alpha/beta IFN (IFN-alpha/beta) completely blocks viral GFP expression. Prior transfection of plasmid DNA induces an IFN response in CEFs and blocks NDV-GFP replication. However, transfection of known inhibitors of the IFN-alpha/beta system, including the influenza A virus NSI protein and the Ebola virus VP35 protein, restores NDV-GFP replication. We therefore conclude that the NDV-GFP virus could be used to screen proteins expressed from plasmids for the ability to counteract the host cell IFN response. Using this system, we show that expression of the NDV V protein or the Nipah virus V, W, or C proteins rescues NDV-GFP replication in the face of the transfection-induced IFN response. The V and W proteins of Nipah virus, a highly lethal pathogen in humans, also block activation of an IFN-inducible promoter in primate cells. Interestingly, the amino-terminal region of the Nipah virus V protein, which is identical to the amino terminus of Nipah virus W, is sufficient to exert the IFN-antagonist activity. In contrast, the anti-IFN activity of the NDV V protein appears to be located in the carboxy-terminal region of the protein, a region implicated in the IFN-antagonist activity exhibited by the V proteins of mumps virus and human parainfluenza virus type 2.