Targeting Outer Membrane β-barrel Proteins of Burkholderia mallei to design a Multi-epitope Vaccine Against Human Glanders

Abstract

Burkholderia mallei, a facultative intracellular Gram-negative pathogen, is the causative agent of glanders that primarily affects solipeds and is sporadically transmitted to humans. Current interventions mainly rely on antibiotics; however, increasing antimicrobial resistance and the lack of a licensed vaccine further complicate disease management. Surface exposed outer membrane {beta}-barrel (OMBB) proteins serve as excellent targets for vaccine development. In the present study, a consensus-based computational framework was employed on the B. mallei turkey2 proteome that identified 59 OMBB proteins - including porins, TonB receptors, autotransporters, and efflux components. These OMBB proteins were leveraged to predict B- and T-cell epitopes which were manually curated, and mapped onto the corresponding protein models to identify surface-exposed epitopes with direct accessibility to the host immune cells. These epitopes were linked together to construct a multi-epitope vaccine (MEV) that was predicted to be antigenic, and soluble upon overexpression. The tertiary structure of the MEV was generated which was used for molecular docking with TLR4 and TLR2. Molecular dynamics simulation and flexibility analysis confirmed the structural stability of the MEV-TLR4/TLR2 complexes. In-silico immune simulation showed the capability of MEV to induce a strong immune response. Codon optimization and in-silico cloning were performed to evaluate its efficient expression in the E. coli host. The findings suggest that surface exposed OMBB proteins can serve as promising antigenic candidates for designing an MEV construct.