Cervantes Rincon, T.; Frckova, T.; Contejean, Z. I.; Cantergiani, J.; Groen, K.; Cena, B.; Moro, S. G.; Bianchini, F.; Simonelli, L.; Jarrossay, D.; Tosolini, S.; Kuratli, R.; Robinson, A. R. E.; Cizkova, M.; Niejadlik, E. G.; Moritz, J.; Thakur, R.; Kratka, Z.; Mijatovic, D.; Grujic, J.; Holoubek, J.; Budakov-Obradovic, Z.; Salat, J.; Honig, V.; Vranes, M.; Lojpur, Z.; Lendak, D.; Sevic, S.; Bajci, M.; Popovic-Dragonjic, L.; Popovska Jovicic, B.; Gavrilovic, J.; Kapoor, T.; MacDonald, M. R.; Bournazos, S.; Varani, L.; Palus, M.; Hale, B. G.; Banovic, P.; Ruzek, D.; Barnes, C. O.; Robbiani, D.

West Nile virus (WNV) is a mosquito-borne pathogen of global concern that can cause fatal neuroinvasive disease. No specific prophylaxis or treatment exists for WNV or related orthoflavivirus infections, and the determinants of human disease severity remain poorly understood. Here, we report that neutralizing autoantibodies against type I interferons do not impair antiviral antibody development. Among the fully human monoclonal antibodies with potent neutralizing activity against WNV that were discovered, W010 targets a unique epitope within the envelope protein domain III (EDIII) and confers both pre- and post-exposure protection in a murine WNV model, even when interferon signaling is impaired. A second protective antibody, W014, exhibits broad cross-neutralization of other pathogenic orthoflavivirus members, including Japanese encephalitis virus, Murray Valley encephalitis virus, Saint Louis encephalitis virus, and Usutu virus. These findings identify key neutralizing epitopes on WNV EDIII and provide candidates for the development of antibody-based interventions against encephalitic orthoflavivirus infections.