Ferrari Alessandro, Cassaniti Irene, Rovida Francesca, Lilleri Daniele, Croce Stefania, Trespidi Francesca, Ghirardello Stefano, Gervais Adrian, Zhang Shen-Ying, Casanova Jean-Laurent, Borghesi Alessandro, Baldanti Fausto
National PhD Programme in One Health approaches to infectious diseases and life science research, Department of Public Health, Experimental and Forensic Medicine, University of Pavia, Pavia, Italy.
Microbiology and Virology Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
Sci Rep. 2025 Apr 2;15(1):11271. doi: 10.1038/s41598-025-89312-6.
Auto-antibodies (auto-Abs) that neutralize type I interferons (IFNs) have been implicated in severe viral infections, including ~ 40% of cases of West Nile virus (WNV) neuroinvasive disease (WNND). Developing robust in vitro models to evaluate the protective effects of type I IFNs against viral infection, as well as the disruptive effects of auto-Abs, is essential for understanding disease pathogenesis and identifying patients at risk. In this study, we used Vero E6 and ARPE-19 cell lines to investigate the ability of type I (IFN-α, IFN-β, IFN-ω), type II (IFN-γ), and type III (IFN-λ1) IFNs to restrict WNV infection. Our results demonstrate that IFN-α, IFN-β, and IFN-ω effectively protect ARPE-19 cells from WNV infection, with IFN-β exhibiting the strongest antiviral effect. In contrast, Vero E6 cells required higher concentrations of IFN-ω to achieve comparable protection. Neither IFN-γ nor IFN-λ1 conferred protection in either cell line. We further screened serum samples from WNV-infected patients for auto-Abs neutralizing type I IFNs. Our findings confirm that the ARPE-19-based assay is consistent with other established methods for detecting neutralizing auto-Abs against type I IFNs. This simple and reliable assay offers a valuable tool for assessing the antiviral effects of type I IFNs and the neutralizing activity of auto-Abs in both research and clinical settings. Future studies should aim to validate the clinical utility of the ARPE-19-WNV infection model on a larger scale.
中和I型干扰素（IFN）的自身抗体（自身抗体）与严重病毒感染有关，包括约40%的西尼罗河病毒（WNV）神经侵袭性疾病（WNND）病例。开发强大的体外模型来评估I型干扰素对病毒感染的保护作用以及自身抗体的破坏作用，对于理解疾病发病机制和识别有风险的患者至关重要。在本研究中，我们使用Vero E6和ARPE-19细胞系来研究I型（IFN-α、IFN-β、IFN-ω）、II型（IFN-γ）和III型（IFN-λ1）干扰素限制WNV感染的能力。我们的结果表明，IFN-α、IFN-β和IFN-ω能有效保护ARPE-19细胞免受WNV感染，其中IFN-β表现出最强的抗病毒效果。相比之下，Vero E6细胞需要更高浓度的IFN-ω才能获得类似的保护。IFN-γ和IFN-λ1在两种细胞系中均未提供保护作用。我们进一步筛选了WNV感染患者的血清样本中的中和I型干扰素的自身抗体。我们的研究结果证实，基于ARPE-19的检测方法与其他检测针对I型干扰素的中和自身抗体的既定方法一致。这种简单可靠的检测方法为评估I型干扰素的抗病毒作用和自身抗体在研究和临床环境中的中和活性提供了一个有价值的工具。未来的研究应旨在更大规模地验证ARPE-19-WNV感染模型的临床实用性。
