摘要Background:West Nile virus is a severe zoonotic pathogen that can cause severe central nervous system symptoms in humans and horses,and is fatal for birds,chickens and other poultry.With no specific drugs or vaccines available,antibody-based therapy is a promising treatment.This study aims to develop neutralizing antibodies against West Nile virus and assess their cross-protective potential against Japanese encephalitis virus.Methods:Monoclonal antibodies against WNV and JEV were isolated by hybridoma technology.The therapeutic efficacy of these antibodies was evaluated using a mouse model,and a humanized version of the monoclonal antibody was generated for potential human application.Results:In this study,we generated eight monoclonal antibodies that exhibit neutralizing activity against WNV.Their therapeutic effects against WNV were validated both in vivo and in vitro.Among these antibodies,C9-G11-F3 also exhibited cross-protective activity against JEV.We also humanized the antibody to ensure that it could be used for WNV infection treatment in humans.Conclusion:This study highlights the importance of neutralizing antibodies as a promising approach for pro-tection against West Nile virus infection and suggests their potential utility in the development of therapeutic interventions.