Computational formulation of a broad-spectrum multi-epitope vaccine against bacterial pathogens implicated in...

Periodontitis is a chronic inflammatory disease driven by dysbiosis of the oral microbiome and is associated with both oral and systemic complications. Key pathogens from the red and orange complexes, along with Chlamydia pneumoniae, contribute significantly to disease progression and related systemic disorders. In this study, emerging biotechnological approaches, including immunoinformatics-driven vaccine design, were employed to develop a multi-epitope vaccine candidate (MEVC) targeting these polymicrobial infections. The MEVC was constructed using 13 B-cell epitopes, 15 cytotoxic T lymphocyte (CTL) epitopes, and 14 helper T lymphocyte (HTL) epitopes identified through experimental evidence and computational prediction. Immunostimulatory linkers and cholera toxin subunit B were incorporated as an adjuvant to enhance immunogenicity. Molecular docking demonstrated strong binding affinities between T-cell epitopes and HLA alleles. Physicochemical analysis indicated that the MEVC is stable, soluble, and exhibits a favourable half-life across biological systems. The construct ws predicted to be antigenic, non-allergenic, and host-compatible. Population coverage analysis of the selected HLA alleles indicated broad global applicability. The tertiary structure of the MEVC was modelled, refined, and docked with TLR2. HADDOCK 2.4 server yielded a binding score of - 196.2 ± 0.0, while PRODIGY predicted a binding affinity of - 13.2 kcal/mol for the MEVC-TLR2 complex. Codon optimization and in silico cloning into the pET-28(+) vector confirmed suitability for expression in Escherichia coli. Molecular dynamics simulations indicated stability of the MEVC-TLR2 complex, while immune simulations predicted strong humoral and cellular responses with sustained IgG, IFN-γ, and IL-2 production upon injection of MEVC into the host. Overall, the MEVC represents a promising therapeutic candidate warranting further experimental validation.

The online version contains supplementary material available at 10.1007/s13205-026-04958-x.