Sensing and Control of Bluetongue Virus Infection in Epithelial Cells via RIG-I and MDA5 Helicases
Sensing and Control of Bluetongue Virus Infection in Epithelial Cells via RIG-I and MDA5 Helicases
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DOI：
10.1128/jvi.00430-12
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发表时间：
2012-11-01
影响因子：
5.4
通讯作者：
Vitour, Damien
中科院分区：
文献类型：
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作者：
Chauveau, Emilie;Doceul, Virginie;Vitour, Damien
蓝舌病毒（BTV）是呼肠孤病毒科的一种节肢动物传播成员，是一种双链RNA病毒，可引发一种在经济上具有重要影响的家畜疾病，近几十年来已在欧洲蔓延。据报道，在BTV感染后，体内和体外都会产生I型干扰素（α/β干扰素[IFN -α/β]）。然而，这一过程中所涉及的细胞传感器和信号通路仍然未知。在此我们研究了针对蓝舌病毒血清型8产生IFN -β的机制。在A549细胞被BTV感染后，IFN -β和其他促炎细胞因子在蛋白质和mRNA水平上都被强烈诱导表达。这种反应似乎依赖于病毒复制，因为暴露于紫外线灭活的病毒无法诱导IFN -β。我们还证明了BTV感染激活了转录因子干扰素调节因子3和核因子κB。我们研究了几种模式识别受体在这种反应中的作用，并表明在通过小干扰RNA介导的视黄酸诱导基因I（RIG - I）或黑色素瘤分化相关基因5（MDA5）编码的RNA解旋酶敲低后，IFN -β的表达大幅降低。相比之下，沉默MyD88、Toll样受体3或最近描述的DexD/H -盒解旋酶DDX1传感器对IFN -β的诱导没有影响或影响微弱，这表明RIG - I样受体途径专门用于感知BTV。此外，我们还表明，RIG - I或MDA5的过表达会削弱感染的A549细胞中BTV的表达。总体而言，这表明RIG - I和MDA5都有助于识别和控制BTV感染。
Bluetongue virus (BTV), an arthropod-borne member of the Reoviridae family, is a double-stranded RNA virus that causes an economically important livestock disease that has spread across Europe in recent decades. Production of type I interferon (alpha/beta interferon [IFN-alpha/beta]) has been reported in vivo and in vitro upon BTV infection. However, the cellular sensors and signaling pathways involved in this process remain unknown. Here we studied the mechanisms responsible for the production of IFN-beta in response to BTV serotype 8. Upon BTV infection of A549 cells, expression of IFN-beta and other proinflammatory cytokines was strongly induced at both the protein and mRNA levels. This response appeared to be dependent on virus replication, since exposure to UV-inactivated virus failed to induce IFN-beta. We also demonstrated that BTV infection activated the transcription factors IFN regulatory factor 3 and nuclear factor kappa B. We investigated the role of several pattern recognition receptors in this response and showed that expression of IFN-beta was greatly reduced after small-interfering-RNA-mediated knockdown of the RNA helicase encoded by retinoic acid-inducible gene I (RIG-I) or melanoma differentiation-associated gene 5 (MDA5). In contrast, silencing of MyD88, Toll-like receptor 3, or the recently described DexD/H-box helicase DDX1 sensor had no or a weak effect on IFN-beta induction, suggesting that the RIG-I-like receptor pathway is specifically engaged for BTV sensing. Moreover, we also showed that overexpression of either RIG-I or MDA5 impaired BTV expression in infected A549 cells. Overall, this indicates that RIG-I and MDA5 can both contribute to the recognition and control of BTV infection.