Developing a universal multi-epitope protein vaccine candidate for enhanced borna virus pandemic preparedness

Introduction: Borna disease virus 1 (BoDV-1) is an emerging zoonotic RNA virus that can cause severe acute encephalitis with high mortality. Currently, there are no effective countermeasures, and the potential risk of a future outbreak requires urgent attention. To address this challenge, the complete genome sequence of BoDV-1 was utilized, and immunoinformatics was applied to identify antigenic peptides suitable for vaccine development.

Methods: Immunoinformatics and antigenicity-focused protein screening were employed to predict B-cell linear epitopes, B-cell conformational epitopes, and cytotoxic T lymphocyte (CTL) epitopes. Only overlapping epitopes with antigenicity greater than 1 and non-toxic, non-allergenic properties were selected for subsequent vaccine construction. The epitopes were linked using GPGPG linkers, incorporating β-defensins at the N-terminus to enhance immune response, and incorporating Hit-6 at the C-terminus to improve protein solubility and aid in protein purification. Computational tools were used to predict the immunogenicity, physicochemical properties, and structural stability of the vaccine. Molecular docking was performed to predict the stability and dynamics of the vaccine in complex with Toll-like receptor 4 (TLR-4) and major histocompatibility complex I (MHC I) receptors. The vaccine construct was cloned through in silico restriction to create a plasmid for expression in a suitable host.

Results: Among the six BoDV-1 proteins analyzed, five exhibited high antigenicity scores. From these, eight non-toxic, non-allergenic overlapping epitopes with antigenicity scores greater than 1 were selected for vaccine development. Computational predictions indicated favorable immunogenicity, physicochemical properties, and structural stability. Molecular docking analysis showed that the vaccine remained stable in complex with TLR-4 and MHC I receptors, suggesting strong potential for immune recognition. A plasmid construct was successfully generated, providing a foundation for the experimental validation of vaccines in future pandemic scenarios.

Discussion: These findings demonstrate the potential of the immunoinformatics-designed multi-epitope vaccines for the prevention and treatment of BoDV-1. Relevant preparations were made in advance for possible future outbreaks and could be quickly utilized for experimental verification.