Abstract
Human metapneumovirus (HMPV) is a globally distributed respiratory virus that can infect individuals of all ages. Infections can lead to severe respiratory illnesses, particularly in young children, the elderly, and immunocompromised individuals. Due to the genetic variability of HMPV and the absence of a vaccine, there is an urgent need for effective vaccine development. This study employed in silico immunoinformatics approaches to design a multi-epitope vaccine targeting the conserved regions fusion glycoprotein (F) of HMPV. A total of 18 highly conserved, antigenic, non-toxic, and non-allergenic epitopes were identified, including 5 B-cell epitopes, 8 cytotoxic T lymphocyte (CTL) epitopes, and 5 helper T lymphocyte (HTL) epitopes. These epitopes were linked using appropriate linkers, and an adjuvant was incorporated to enhance immunogenicity. Computational analyses predicted the vaccine construct to be antigenic, non-allergenic, and non-toxic. Molecular docking studies with Toll-like receptor 4 (TLR4) demonstrated a strong binding affinity, with a binding energy of - 112.7 ± 9.7 kcal/mol and a Z-score of - 2.5. Molecular dynamics simulations further confirmed the stability of the vaccine-TLR4 complex. Immune simulation analysis predicted robust immune responses, including elevated levels of IgM, IgG1, IgG2, and combined IgG + IgM. Finally, in silico codon optimization and cloning analysis indicated that the vaccine construct could be efficiently expressed in E. coli. These findings support the potential of the proposed multi-epitope vaccine as a candidate for HMPV prevention; however, experimental in vitro and in vivo studies are required to validate its immunogenicity, safety, and efficacy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13337-025-00932-y.