From Epitope Discovery to Immune Simulation: An Integrated Computational Strategy for Multi-Epitope Vaccine Design Against Human Mastadenovirus

Abstract

Human Adenovirus (ADV), a member of the Adenoviridae family, is Double-Stranded DNA (DSDNA) virus associated with gastroenteritis, conjunctivitis, and severe respiratory infection and illnesses in infants and immunocompromised individuals. Despite its widespread prevalence in food and water sources, no specific and effective treatment strategy is currently available. In the current study, robust immunoinformatic approaches were employed to develop a multi-epitope vaccine targeting conserved fiber protein of adenovirus. Non-allergic and antigenic and non-toxic B-cell, Cytotoxic T Lymphocyte (CTL), and Helper T Lymphocyte (HTL) epitopes were predicted and combined with appropriate linkers, such as KK, GPGPG, and AAY. Additionally, human beta-defensin was added as an adjuvant at the C-terminal end of the vaccine construct using the EAAAK linker to enhance immunogenicity. The designed vaccine exhibited favorable physicochemical properties with a solubility score of 0.8327, and an Antigenicity potential score 0.913813, while the construct remained non-allergenic and non-toxic. Structural validation through RAMPAGE, ProSa web, and ERRAT confirmed the reliability of the vaccine model, with an ERRAT value 86.475 and 91.3% residues located in the most favored region of the Ramachandran plot. The vaccine also showed a ProSA Z-score of -6.87, further indicated structural stability and quality. Molecular docking analysis revealed strong interactions between the vaccine construct and Toll-like receptor 2 (TLR-2 with a binding energy score of −257.02 kJ/mol. Subsequently, Molecular dynamics simulations further confirmed the stability of the vaccine TLR-2 complex. Codon optimization and in-silico cloning into the pET28a (+) vector suggested efficient expression potential in E. coli. Additionally, immune simulation studies predicted a promising immunological response. Collectively, the proposed multi-epitope vaccine construct demonstrated promising immunogenic and structural characteristics against adenovirus infection. However, future in vitro and in vivo experimental studies are required to confirm its safety, efficacy, and therapeutic potential for preventing and treating of adenovirus-related diseases.