Development of a novel next-generation multi-epitope vaccine targeting the Guaroa virus glycoprotein precursor through reverse vaccinology

This study aimed to develop a multi-epitope vaccine candidate against Guaroa virus using integrated immunoinformatics and reverse vaccinology approaches. The viral glycoprotein precursor was selected as the target for epitope screening. Potential B-cell and T-cell epitopes with predicted antigenic properties, non-allergenicity, and non-toxicity were identified through computational analyses. Global population coverage assessment predicted a population coverage of 99.09% based on the selected HLA-associated epitopes. Selected epitopes were assembled into a chimeric vaccine construct containing β-defensin-3 adjuvant and linker sequences. Physicochemical analysis predicted properties relevant to solubility and recombinant expression, although these characteristics require experimental assessment. Structural prediction and refinement yielded a model in which 99.5% of residues were located within favored regions. Molecular docking predicted interactions between the vaccine construct and immune receptors TLR2, TLR3, and TLR4, with TLR4 showing the lowest docking score among the three receptors. Molecular dynamics simulations were used to assess the predicted structural behavior of the vaccine-receptor complexes. The vaccine-TLR4 complex yielded a docking score of -1212.9, while MM-GBSA analysis produced a ΔG_bind estimate of -227.14 kcal/mol based on the final trajectory frame. Immune simulation predicted increases in antibody production, cytokine release, and memory cell formation. Collectively, these findings support the computational prioritization of the construct as a vaccine candidate for further experimental investigation, although experimental and clinical validation remain necessary.