Characterization of Outer Membrane β-barrel Proteins of Pasteurella multocida for Multi-Epitope Vaccine Design against Human Pasteurellosis

Abstract

Pasteurella multocida is a facultative anaerobic, Gram-negative coccobacillus that causes pasteurellosis in companion animals, livestock, and poultry and poses a significant zoonotic risk to humans through bite wounds, scratches, licking, and transfer of bodily fluids. Although vaccines are available for livestock and poultry, no vaccine is currently licensed for human use. In this study, we systematically identified and characterized 29 outer membrane {beta}-barrel (OMBB) proteins in P. multocida Past9 proteome and classified them into functional categories, including TonB-dependent receptors, porins, autotransporters, adhesins, and efflux pumps. B-cell, cytotoxic T-lymphocyte (CTL), and helper T-lymphocyte (HTL) epitopes were predicted from the identified proteins and screened based on antigenicity, non-allergenicity, and non-toxicity. Epitopes conserved across eight human-infecting P. multocida strains and located within the extracellular loop (ECL) region were incorporated into a multi-epitope vaccine (MEV) construct. The designed MEV was predicted to be antigenic, non-allergenic, and soluble. Its tertiary structural model was iteratively refined and validated. Molecular docking with human toll-like receptors 4/2 (TLR4/TLR2) predicted stable interactions, further supported by 100 ns molecular dynamics simulations. Immune simulation of the MEV construct predicted a strong simulated immune response. Furthermore, codon optimization and in silico cloning supported the feasibility of recombinant MEV expression in E. coli. The construct was benchmarked against OmpH, a well-known antigenic protein and exhibited broadly comparable predicted immune responses and receptor-binding energetics. This study proposes a designed MEV candidate against human pasteurellosis and highlights OMBB proteins as potential immunogenic targets for vaccine development.