Evaluation of the immunogenic properties of a chimeric protein against West Nile virus: in-silico analysis and assessment in mouse

Abstract

Background and Objectives: West Nile virus (WNV) is a mosquito-borne flavivirus that represents a significant public health concern, and the development of effective vaccine candidates remains a priority. This study aimed to design a chimeric multi-epitope protein against WNV using immunoinformatics approaches and evaluate its immunogenicity in mice.

Materials and Methods: Selected regions of the WNV envelope (E) and non-structural 1 (NS1) proteins were analyzed to identify B- and T-cell epitopes using immunoinformatics tools. The selected epitopes were incorporated into a chimeric con- struct fused with cholera toxin B subunit, and its antigenicity, allergenicity, structural properties, and TLR3 interaction were evaluated in silico. The optimized construct was expressed in Escherichia coli, purified by Ni-NTA affinity chromatography, and confirmed by SDS-PAGE and western blotting. BALB/c mice were immunized with the recombinant protein plus adju- vant, and immune responses were assessed using ELISA, cytokine assays, and flow cytometry.

Results: Immunized mice developed high IgG antibody titers and showed detectable IFN-γ and IL-5 production, suggesting induction of Th1-associated responses with limited Th2-related cytokine production. The chimeric protein also stimulated CD4+ and CD8+ T-cell responses, indicating activation of both humoral and cellular immunity.

Conclusion: The designed chimeric protein showed immunogenic potential and induced humoral and cellular immune re-sponses in mice, supporting further evaluation as a candidate for WNV vaccine development.