Evaluation of the immunogenic properties of a chimeric protein against West Nile virus: in-silico analysis and assessment in mouse
- 1Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
- 2Department of Arboviruses and Viral Hemorrhagic Fevers, National Reference Laboratory, Pasteur Institute of Iran, Tehran, Iran
- 3Department of Plant Molecular Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
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 construct 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 adjuvant, 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 responses in mice, supporting further evaluation as a candidate for WNV vaccine development.
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BibTeX
@article{arezoumandi2026evaluation,
title = {{Evaluation of the immunogenic properties of a chimeric protein against West Nile virus: in-silico analysis and assessment in mouse}},
author = {Mahdi Arezoumandi and Sadegh Shojaei Baghini and Nima Rad and Elham Taghipour and Mohammad Hassan Pouriayevali and Fateme Frootan and Mostafa Motallebi and Ali Hatef Salmanian},
journal = {Iranian Journal of Microbiology},
year = {2026},
month = nov,
doi = {10.18502/ijm.v18i5.22876},
url = {https://doi.org/10.18502/ijm.v18i5.22876},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Evaluation of the immunogenic properties of a chimeric protein against West Nile virus: in-silico analysis and assessment in mouse
AU - Mahdi Arezoumandi
AU - Sadegh Shojaei Baghini
AU - Nima Rad
AU - Elham Taghipour
AU - Mohammad Hassan Pouriayevali
AU - Fateme Frootan
AU - Mostafa Motallebi
AU - Ali Hatef Salmanian
PY - 2026
DA - 2026/11/06
JO - Iranian Journal of Microbiology
DO - 10.18502/ijm.v18i5.22876
UR - https://doi.org/10.18502/ijm.v18i5.22876
AB - 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 construct 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 adjuvant, 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 responses in mice, supporting further evaluation as a candidate for WNV vaccine development.
SN - 2008-3289
SN - 2008-4447
ER -