Gandolfo Claudia, Terrosi Chiara, Prathyumnan Shibily, Anichini Gabriele, Savellini Gianni Gori, Morgante Giuseppe, Cusi Maria Grazia
Department of Medical Biotechnologies, University of Siena, 53100 Siena, Italy.
Department of Molecular and Developmental Medicine, University of Siena, 53100 Siena, Italy.
Pathogens. 2022 Mar 5;11(3):321. doi: 10.3390/pathogens11030321.
The rapid spread of new outbreaks of human infection caused by Zika virus (ZIKV) has raised many global concerns since 2016. Despite the increasing knowledge of this virus, data on the pathogenesis of ZIKV are still missing. In particular, it is still unknown how the virus crosses the endothelial monolayer and gets access to the bloodstream. In the present work, we used human umbilical vein endothelial cells (HUVECs) as a model to study ZIKV infection . We demonstrated that HUVECs are an optimal reservoir for viral replication, as they were able to sustain ZIKV infection up to two weeks, without showing a cytopathic effect. In order to evaluate the integrity of endothelial monolayer, immunofluorescence was performed on mock-infected or ZIKV-infected cells ± peripheral blood mononuclear cells (PBMCs) or polymorphonuclear cells (PMN), 48 h p.i., by using an anti-VE-Cadherin antibody, a major adherence protein that maintains the integrity of intercellular junctions. In addition to infection, we noted that the presence of some components of the immune system, such as PMNs, played an important role in altering the endothelial monolayer in cell junctions, suggesting that presence at the site of infection probably promotes the spread of ZIKV in vivo in the bloodstream.
自2016年以来，寨卡病毒（ZIKV）引发的人类感染新疫情迅速蔓延，引发了全球诸多关注。尽管对这种病毒的了解不断增加，但关于ZIKV发病机制的数据仍然缺失。特别是，病毒如何穿过内皮单层并进入血液循环仍不清楚。在本研究中，我们使用人脐静脉内皮细胞（HUVECs）作为模型来研究ZIKV感染。我们证明HUVECs是病毒复制的最佳储存库，因为它们能够维持ZIKV感染长达两周，而不表现出细胞病变效应。为了评估内皮单层的完整性，在感染后48小时，使用抗VE-钙黏蛋白抗体（一种维持细胞间连接完整性的主要黏附蛋白），对模拟感染或ZIKV感染的细胞±外周血单核细胞（PBMCs）或多形核细胞（PMN）进行免疫荧光检测。除了感染，我们还注意到免疫系统的一些成分，如PMN，在改变细胞连接处的内皮单层方面发挥了重要作用，这表明它们在感染部位的存在可能促进了ZIKV在体内血液中的传播。
