Visualization of Double-Stranded RNA Colocalizing With Pattern Recognition Receptors in Arenavirus Infected Cells.
Visualization of Double-Stranded RNA Colocalizing With Pattern Recognition Receptors in Arenavirus Infected Cells.
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DOI：
10.3389/fcimb.2018.00251
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发表时间：
2018
影响因子：
5.7
通讯作者：
Huang C
中科院分区：
文献类型：
--
作者：
Mateer EJ;Paessler S;Huang C
启动病毒感染先天免疫应答的一个重要步骤是细胞质模式识别受体（PRRs）识别非自体病毒RNA，包括双链RNA （dsRNA）。对于许多正义RNA病毒和DNA病毒，病毒dsRNA的产生以及病毒dsRNA与PRRs的相互作用都被很好地描述了。然而，对于负义RNA病毒，病毒dsRNA被认为是在低到不可检测的水平上产生的，并且病毒dsRNA的PRR识别在很大程度上仍然不清楚。在沙粒病毒的情况下，核鞘蛋白（NP）已被确定含有一种外核糖核酸酶活性，在生化研究中优先降解dsRNA。然而，致病性新世界（NW）沙粒病毒感染很容易以rig - 1依赖的方式诱导干扰素（IFN）反应，并激活dsrna依赖的蛋白激酶R （PKR）。为了更好地了解致病性沙粒病毒感染的先天免疫反应，我们使用了一种新发现的dsRNA特异性抗体，该抗体可以有效地检测负义RNA病毒感染细胞中的病毒dsRNA。免疫荧光法检测了NW沙粒病毒感染细胞与病毒NP共定位的dsRNA。重要的是，dsRNA信号还与细胞质PRRs，即PKR、RIG-I和MDA-5，以及感染细胞中PKR的磷酸化活化形式共定位。我们的数据清楚地证明了在NW沙粒病毒感染的细胞中，PRR对dsRNA的识别及其激活。这些发现为NW沙粒病毒与宿主先天免疫反应之间的相互作用提供了新的见解。
An important step in the initiation of the innate immune response to virus infection is the recognition of non-self, viral RNA, including double-stranded RNA (dsRNA), by cytoplasmic pattern recognition receptors (PRRs). For many positive-sense RNA viruses and DNA viruses, the production of viral dsRNA, and the interaction of viral dsRNA and PRRs are well characterized. However, for negative-sense RNA viruses, viral dsRNA was thought to be produced at low to undetectable levels and PRR recognition of viral dsRNA is still largely unclear. In the case of arenaviruses, the nucleocaspid protein (NP) has been identified to contain an exoribonuclease activity that preferentially degrades dsRNA in biochemical studies. Nevertheless, pathogenic New World (NW) arenavirus infections readily induce an interferon (IFN) response in a RIG-I dependent manner, and also activate the dsRNA-dependent Protein Kinase R (PKR). To better understand the innate immune response to pathogenic arenavirus infection, we used a newly identified dsRNA-specific antibody that efficiently detects viral dsRNA in negative-sense RNA virus infected cells. dsRNA was detected in NW arenavirus infected cells colocalizing with virus NP in immunofluorescence assay. Importantly, the dsRNA signals also colocalized with cytoplasmic PRRs, namely, PKR, RIG-I and MDA-5, as well as with the phosphorylated, activated form of PKR in infected cells. Our data clearly demonstrate the PRR recognition of dsRNA and their activation in NW arenavirus infected cells. These findings provide new insights into the interaction between NW arenaviruses and the host innate immune response.