Cell-to-Cell Contact and Nectin-4 Govern Spread of Measles Virus from Primary Human Myeloid Cells to Primary Human Airway Epithelial Cells.
Cell-to-Cell Contact and Nectin-4 Govern Spread of Measles Virus from Primary Human Myeloid Cells to Primary Human Airway Epithelial Cells.
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细胞间接触和 Nectin-4 控制麻疹病毒从原代人骨髓细胞传播到原代人气道上皮细胞。
DOI：
10.1128/jvi.00266-16
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发表时间：
2016
影响因子：
5.4
通讯作者：
Sinn,PatrickL
中科院分区：
文献类型：
--
作者：
Singh,BrajeshK;Li,Ni;Mark,AnnaC;Mateo,Mathieu;Cattaneo,Roberto;Sinn,PatrickL
麻疹是一种传染性极强的急性病毒性疾病。气道内的免疫细胞可能是感染的第一目标，这些细胞将麻疹病毒（MeV）运输到淋巴结进行扩增和随后的全身传播。被感染的免疫细胞被认为将MeV返回到气道;然而，对病毒转移到肺上皮细胞的机制知之甚少。为了研究这一过程，我们收集了来自人类供体的血液，并产生了原代骨髓细胞，特别是单核细胞衍生的巨噬细胞（MDM）和树突状细胞（DC）。用MeV感染MDM和DC，然后将其应用于来自人供体（HAE）的分化良好的气道上皮细胞的原代培养物。与先前用游离病毒获得的结果一致，当应用于顶端表面时，感染的MDM或DC不能将MeV转移至HAE。同样，将感染的MDM或DC应用于在小孔（0.4 μm）支持膜上生长的HAE基底外侧表面，也不会转移病毒。相比之下，感染的MDM和DC应用于在大孔（3.0 μm）膜上生长的HAE的基底外侧表面成功地转移MeV。共聚焦显微镜显示MDM和DC能够穿透大孔膜，但不能穿透小孔膜。此外，通过使用连接蛋白-4阻断抗体或不能通过连接蛋白-4进入细胞的重组MeV，我们正式证明了从免疫细胞到HAE的转移以连接蛋白-4依赖性方式发生。因此，受感染的MDM和DC都依赖于细胞与细胞的接触和nectin-4来有效地将MeV递送到HAE的基底外侧表面。这种快速传播是基于细胞与细胞的接触，而不是颗粒的释放和重返。在这里，我们证明MeV从感染的免疫细胞转移到上皮细胞也发生细胞与细胞的接触，而不是通过无细胞颗粒。此外，我们试图确定哪些免疫细胞将MeV感染性转移到人气道上皮。我们的研究基于两种类型的人原代细胞：（i）从捐献的血液中产生的骨髓细胞和（ii）来自供体肺的分化良好的气道上皮细胞。我们发现，不同类型的骨髓细胞，即，单核细胞衍生的巨噬细胞和树突状细胞，将感染转移到气道上皮细胞。此外，细胞与细胞的接触是成功转移MeV的重要组成部分。我们的研究阐明了最具传染性的人类呼吸道病毒传递到气道上皮的机制。
Measles is a highly contagious, acute viral illness. Immune cells within the airways are likely first targets of infection, and these cells traffic measles virus (MeV) to lymph nodes for amplification and subsequent systemic dissemination. Infected immune cells are thought to return MeV to the airways; however, the mechanisms responsible for virus transfer to pulmonary epithelial cells are poorly understood. To investigate this process, we collected blood from human donors and generated primary myeloid cells, specifically, monocyte-derived macrophages (MDMs) and dendritic cells (DCs). MDMs and DCs were infected with MeV and then applied to primary cultures of well-differentiated airway epithelial cells from human donors (HAE). Consistent with previous results obtained with free virus, infected MDMs or DCs were incapable of transferring MeV to HAE when applied to the apical surface. Likewise, infected MDMs or DCs applied to the basolateral surface of HAE grown on small-pore (0.4-μm) support membranes did not transfer virus. In contrast, infected MDMs and DCs applied to the basolateral surface of HAE grown on large-pore (3.0-μm) membranes successfully transferred MeV. Confocal microscopy demonstrated that MDMs and DCs are capable of penetrating large-pore membranes but not small-pore membranes. Further, by using a nectin-4 blocking antibody or recombinant MeV unable to enter cells through nectin-4, we demonstrated formally that transfer from immune cells to HAE occurs in a nectin-4-dependent manner. Thus, both infected MDMs and DCs rely on cell-to-cell contacts and nectin-4 to efficiently deliver MeV to the basolateral surface of HAE.IMPORTANCEMeasles virus spreads rapidly and efficiently in human airway epithelial cells. This rapid spread is based on cell-to-cell contact rather than on particle release and reentry. Here we posit that MeV transfer from infected immune cells to epithelial cells also occurs by cell-to-cell contact rather than through cell-free particles. In addition, we sought to determine which immune cells transfer MeV infectivity to the human airway epithelium. Our studies are based on two types of human primary cells: (i) myeloid cells generated from donated blood and (ii) well-differentiated airway epithelial cells derived from donor lungs. We show that different types of myeloid cells, i.e., monocyte-derived macrophages and dendritic cells, transfer infection to airway epithelial cells. Furthermore, cell-to-cell contact is an important component of successful MeV transfer. Our studies elucidate a mechanism by which the most contagious human respiratory virus is delivered to the airway epithelium.