KU Leuven Department of Microbiology, Immunology and Transplantation, Rega Institute, Laboratory of Virology and Chemotherapy, Leuven, Belgium.
Molecular Vaccinology and Vaccine Discovery group, Leuven, Belgium.
Nat Commun. 2024 Jan 2;15(1):42. doi: 10.1038/s41467-023-44339-z.
To curb viral epidemics and pandemics, antiviral drugs are needed with activity against entire genera or families of viruses. Here, we develop a cell-based multiplex antiviral assay for high-throughput screening against multiple viruses at once, as demonstrated by using three distantly related orthoflaviviruses: dengue, Japanese encephalitis and yellow fever virus. Each virus is tagged with a distinct fluorescent protein, enabling individual monitoring in cell culture through high-content imaging. Specific antisera and small-molecule inhibitors are employed to validate that multiplexing approach yields comparable inhibition profiles to single-virus infection assays. To facilitate downstream analysis, a kernel is developed to deconvolute and reduce the multidimensional quantitative data to three cartesian coordinates. The methodology is applicable to viruses from different families as exemplified by co-infections with chikungunya, parainfluenza and Bunyamwera viruses. The multiplex approach is expected to facilitate the discovery of broader-spectrum antivirals, as shown in a pilot screen of approximately 1200 drug-like small-molecules.
为了遏制病毒疫情和大流行，我们需要具有针对整个病毒属或病毒科的活性的抗病毒药物。在这里，我们开发了一种基于细胞的多重抗病毒测定法，可一次高通量筛选多种病毒，如使用三种远缘正粘病毒（登革热、日本脑炎和黄热病病毒）所示。每种病毒都标记有独特的荧光蛋白，通过高内涵成像在细胞培养中可单独进行监测。特异性抗血清和小分子抑制剂的使用验证了该多重方法与单病毒感染测定法产生相当的抑制谱。为了便于下游分析，开发了一个内核将多维定量数据解卷积并减少到三个笛卡尔坐标。该方法适用于不同科的病毒，如用基孔肯雅热、副流感和布尼亚韦拉病毒进行共感染的例子。如大约 1200 种类似药物的小分子的初步筛选所示，这种多重方法有望促进更广泛谱抗病毒药物的发现。
