Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies

Abstract 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 v...

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Main Authors: Alessandro Ferrari, Irene Cassaniti, Francesca Rovida, Daniele Lilleri, Stefania Croce, Francesca Trespidi, Stefano Ghirardello, Adrian Gervais, Shen-Ying Zhang, Jean-Laurent Casanova, Alessandro Borghesi, Fausto Baldanti
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Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-89312-6
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author Alessandro Ferrari
Irene Cassaniti
Francesca Rovida
Daniele Lilleri
Stefania Croce
Francesca Trespidi
Stefano Ghirardello
Adrian Gervais
Shen-Ying Zhang
Jean-Laurent Casanova
Alessandro Borghesi
Fausto Baldanti
author_facet Alessandro Ferrari
Irene Cassaniti
Francesca Rovida
Daniele Lilleri
Stefania Croce
Francesca Trespidi
Stefano Ghirardello
Adrian Gervais
Shen-Ying Zhang
Jean-Laurent Casanova
Alessandro Borghesi
Fausto Baldanti
author_sort Alessandro Ferrari
collection DOAJ
description Abstract 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.
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spelling doaj-art-34496ce262bb44ccb294089aa0b4e4742025-08-20T02:08:08ZengNature PortfolioScientific Reports2045-23222025-04-0115111310.1038/s41598-025-89312-6Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodiesAlessandro Ferrari0Irene Cassaniti1Francesca Rovida2Daniele Lilleri3Stefania Croce4Francesca Trespidi5Stefano Ghirardello6Adrian Gervais7Shen-Ying Zhang8Jean-Laurent Casanova9Alessandro Borghesi10Fausto Baldanti11National PhD Programme in One Health approaches to infectious diseases and life science research, Department of Public Health, Experimental and Forensic Medicine, University of PaviaDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of PaviaDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of PaviaMicrobiology and Virology Unit, Fondazione IRCCS Policlinico San MatteoUOSD Cell Factory, San Matteo Research HospitalNeonatal Intensive Care Unit, Fondazione IRCCS Policlinico San MatteoNeonatal Intensive Care Unit, Fondazione IRCCS Policlinico San MatteoLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Institut National de la Santé et de la Recherche Médicale (INSERM) U1163, Necker Hospital for Sick ChildrenLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Institut National de la Santé et de la Recherche Médicale (INSERM) U1163, Necker Hospital for Sick ChildrenLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Institut National de la Santé et de la Recherche Médicale (INSERM) U1163, Necker Hospital for Sick ChildrenNeonatal Intensive Care Unit, Fondazione IRCCS Policlinico San MatteoDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of PaviaAbstract 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.https://doi.org/10.1038/s41598-025-89312-6WNVType I IFNsAuto-antibodyImmunityNeutralizing antibody
spellingShingle Alessandro Ferrari
Irene Cassaniti
Francesca Rovida
Daniele Lilleri
Stefania Croce
Francesca Trespidi
Stefano Ghirardello
Adrian Gervais
Shen-Ying Zhang
Jean-Laurent Casanova
Alessandro Borghesi
Fausto Baldanti
Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies
Scientific Reports
WNV
Type I IFNs
Auto-antibody
Immunity
Neutralizing antibody
title Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies
title_full Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies
title_fullStr Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies
title_full_unstemmed Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies
title_short Human type I interferons protect Vero E6 and ARPE-19 cells against West Nile virus and are neutralized by pathogenic autoantibodies
title_sort human type i interferons protect vero e6 and arpe 19 cells against west nile virus and are neutralized by pathogenic autoantibodies
topic WNV
Type I IFNs
Auto-antibody
Immunity
Neutralizing antibody
url https://doi.org/10.1038/s41598-025-89312-6
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