Katholieke Universiteit Leuven, Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium.
Microbiol Spectr. 2022 Aug 31;10(4):e0049122. doi: 10.1128/spectrum.00491-22. Epub 2022 Jul 11.
In this study, we use electric cell-substrate impedance sensing (ECIS), an established cell-based electrical impedance (CEI) technology, to decipher the kinetic cytopathic effect (CPE) induced by Zika virus (ZIKV) in susceptible human A549 lung epithelial cells and to evaluate several classes of compounds with reported antiviral activity (two entry inhibitors and two replication inhibitors). To validate the assay, we compare the results with those obtained with more traditional methods based on cell viability and viral yield readouts. We demonstrate that CEI can detect viral infection in a sensitive manner and can be used to determine antiviral potency. Moreover, CEI has multiple benefits compared to conventional assays: the technique is less laborious and better at visualizing the dynamic antiviral activity profile of the compounds, while also it has the ability to determine interesting time points that can be selected as endpoints in assays without continuous readout. We describe several parameters to characterize the compounds' cytotoxicity and their antiviral activity profile. In addition, the CEI patterns provide valuable additional information about the presumed mechanism of action of these compounds. Finally, as a proof of concept, we used CEI to evaluate the antiviral activity of a small series of compounds, for which we demonstrate that the sulfonated polymer PRO2000 inhibits ZIKV with a response profile representative for a viral entry inhibitor. Overall, we demonstrate for the first time that CEI is a powerful technology to evaluate and characterize compounds against ZIKV replication in a real-time, label-free, and noninvasive manner. Zika virus can cause serious disease in humans. Unfortunately, no antiviral drugs are available to treat infection. Here, we use an impedance-based method to continuously monitor virus infection in-and damage to-human cells. We can determine the Zika viral dose with this technique and also evaluate whether antiviral compounds protect the cells from damage caused by virus replication. We also show that this technique can be used to further unravel the characteristics of these compounds, such as their toxicity to the cells, and that it might even give further insight in their mechanism of antiviral action. Finally, we also find a novel Zika virus inhibitor, PRO2000. Overall, in this study, we use the impedance technology to-for the first time-evaluate compounds with anti-Zika virus properties, and therefore it can add valuable information in the further search for antiviral drugs.
在这项研究中，我们使用了细胞-基质阻抗感应（ECIS）技术，这是一种已建立的基于细胞的电阻抗（CEI）技术，以破译寨卡病毒（ZIKV）在易感的人 A549 肺上皮细胞中引起的细胞病变效应（CPE），并评估几类具有报道的抗病毒活性的化合物（两种进入抑制剂和两种复制抑制剂）。为了验证该测定法，我们将结果与基于细胞活力和病毒产量读数的更传统方法的结果进行了比较。我们证明 CEI 可以敏感地检测病毒感染，并可用于确定抗病毒效力。此外，与传统测定法相比，CEI 具有多个优点：该技术不那么费力，更善于观察化合物的动态抗病毒活性谱，同时还能够确定有趣的时间点，这些时间点可作为无连续读数的测定法的终点进行选择。我们描述了几个参数来描述化合物的细胞毒性及其抗病毒活性谱。此外，CEI 模式提供了有关这些化合物假定作用机制的有价值的附加信息。最后，作为概念验证，我们使用 CEI 评估了一小系列化合物的抗病毒活性，结果表明磺化聚合物 PRO2000 以代表病毒进入抑制剂的反应谱抑制 ZIKV。总的来说，我们首次证明，CEI 是一种强大的技术，可实时，无标记和非侵入性地评估和表征针对 ZIKV 复制的化合物。寨卡病毒会在人类中引起严重疾病。不幸的是，尚无可用的抗病毒药物来治疗感染。在这里，我们使用基于阻抗的方法连续监测病毒感染和人类细胞的损伤。我们可以用这种技术确定寨卡病毒的剂量，还可以评估抗病毒化合物是否可以保护细胞免受病毒复制引起的损伤。我们还表明，该技术可用于进一步阐明这些化合物的特性，例如它们对细胞的毒性，甚至可能进一步揭示其抗病毒作用机制。最后，我们还发现了一种新的寨卡病毒抑制剂 PRO2000。总的来说，在这项研究中，我们首次使用阻抗技术来评估具有抗寨卡病毒特性的化合物，因此它可以为寻找抗病毒药物提供有价值的信息。
