Measles Virus Ribonucleoprotein Complexes Rapidly Spread across Well-Differentiated Primary Human Airway Epithelial Cells along F-Actin Rings.
Measles Virus Ribonucleoprotein Complexes Rapidly Spread across Well-Differentiated Primary Human Airway Epithelial Cells along F-Actin Rings.
复制标题
麻疹病毒核糖核蛋白复合物沿着 F-肌动蛋白环快速扩散到分化良好的原代人气道上皮细胞中。
DOI：
10.1128/mbio.02434-19
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发表时间：
2019
期刊：
影响因子：
6.4
通讯作者：
Sinn,PatrickL
中科院分区：
文献类型：
--
作者：
Singh,BrajeshK;Pfaller,ChristianK;Cattaneo,Roberto;Sinn,PatrickL
麻疹病毒（MeV）是一种高度传染性的人类病原体，仍然是世界范围内的健康负担。MeV传播研究的挑战之一是识别准确反映MeV在人体中行为的模型系统。在我们的研究中，我们使用来自人供体肺的未传代的、分化良好的气道上皮细胞原代培养物来检查MeV感染和传播。在这里，我们表明，MeV的核糖核蛋白复合物（RNP），核衣壳和磷蛋白的主要成分，共定位与顶端和circumaillary F-肌动蛋白网络。为了更好地了解MeV感染如何在气道上皮中传播，我们产生了一种重组病毒，其RNP复合物中包含嵌合荧光蛋白。通过活细胞成像，我们观察到RNP沿着新感染细胞的环膜F-肌动蛋白环快速移动。这种水平运输穿过上皮细胞的惊人快速机制与病毒膜融合装置打开柱状细胞之间的孔一致。我们的工作提供了机械的见解，如何兆电子伏迅速传播通过气道上皮细胞，有助于其极具传染性的nature.IMPORTANCEThe病毒颗粒的能力，直接传播细胞到细胞内的气道没有颗粒释放被认为是非常有利的许多呼吸道病毒。我们以前在分化良好的原代人气道上皮细胞中的研究表明，麻疹病毒（MeV）通过诱导细胞间膜孔的形成在细胞间传播。基于一个新产生的核糖核蛋白复合物（RNP）的“跟踪”病毒，我们文件活细胞显微镜下，兆电子伏RNP移动沿着F-肌动蛋白环进入一个新的细胞。因此，RNP不是通过新感染的柱状细胞的细胞质扩散，而是利用细胞骨架结构快速横向扩散穿过人气道上皮。这导致通过上皮的快速水平扩散，而不需要颗粒释放。
Measles virus (MeV) is a highly contagious human pathogen that continues to be a worldwide health burden. One of the challenges for the study of MeV spread is the identification of model systems that accurately reflect how MeV behaves in humans. For our studies, we use unpassaged, well-differentiated primary cultures of airway epithelial cells from human donor lungs to examine MeV infection and spread. Here, we show that the main components of the MeV ribonucleoprotein complex (RNP), the nucleocapsid and phosphoprotein, colocalize with the apical and circumapical F-actin networks. To better understand how MeV infections spread across the airway epithelium, we generated a recombinant virus incorporating chimeric fluorescent proteins in its RNP complex. By live cell imaging, we observed rapid movement of RNPs along the circumapical F-actin rings of newly infected cells. This strikingly rapid mechanism of horizontal trafficking across epithelia is consistent with the opening of pores between columnar cells by the viral membrane fusion apparatus. Our work provides mechanistic insights into how MeV rapidly spreads through airway epithelial cells, contributing to its extremely contagious nature.IMPORTANCEThe ability of viral particles to directly spread cell to cell within the airways without particle release is considered to be highly advantageous to many respiratory viruses. Our previous studies in well-differentiated, primary human airway epithelial cells suggest that measles virus (MeV) spreads cell to cell by eliciting the formation of intercellular membrane pores. Based on a newly generated ribonucleoprotein complex (RNP) “tracker” virus, we document by live-cell microscopy that MeV RNPs move along F-actin rings before entering a new cell. Thus, rather than diffusing through the cytoplasm of a newly infected columnar cell, RNPs take advantage of the cytoskeletal infrastructure to rapidly spread laterally across the human airway epithelium. This results in rapid horizontal spread through the epithelium that does not require particle release.