Identification of conserved vaccine targets in Mycoplasma bovis through integrated pan-genome and reverse vaccinology approaches with in vivo immunogenicity and safety evaluation.

Three M. bovis antigens show promise as safe, immunogenic subunit vaccine candidates for bovine respiratory disease.

This study addresses Mycoplasma bovis, a significant pathogen causing bovine respiratory disease and substantial economic losses to the cattle industry. Researchers analyzed 80 global M. bovis genomes to identify 1,058 core genes, then employed reverse vaccinology and immunoinformatics to screen three vaccine candidates: lipoate protein ligase 2 (LplA), elongation factor 4 (LepA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Recombinant proteins were expressed in E. coli and evaluated in 30 BALB/c mice across five groups. Primary immunization with complete Freund's adjuvant was followed by booster doses with incomplete adjuvant at weeks 2 and 4. All vaccinated groups demonstrated significantly elevated antigen-specific antibody levels by day 14, sustained through week 6, with rLplA and rGAPDH showing superior titers. Analysis revealed predominantly Th2-type immune responses, and all candidates substantially increased cytokine production. Critically, no adverse physiological or histopathological effects were observed, establishing excellent safety profiles. These findings present promising subunit vaccine targets addressing pathogen variability, antibiotic resistance, and current vaccine limitations in bovine medicine.

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