Design of B‐Cell Multi‐Epitope Subunit Vaccines Against Glaesserella parasuis by Reverse Vaccinology: An In Silico and In Vivo Study

(GPS) is a severe disease that results in substantial economic losses to the swine industry worldwide. Here we describe a multiepitope vaccine cocktail (MEVC) that was designed using reverse vaccinology and immunoinformatics. The MEVC was comprised of three multiepitope subunits (MESs, designated as TB, 14B, and 24B), which were constructed using 14 B-cell epitopes predicted from six outer membrane antigens of GPS. The MESs exhibited non-allergenicity, high antigenicity, solubility, and stability. Predicted secondary and tertiary structures of the MESs were validated and showed strong binding affinity with the swine leukocyte antigen (SLA) by molecular docking. Immune simulation analysis further confirmed robust induction of both cellular and humoral immune responses. Immunization with MESs plus Gel-01 adjuvant (MEVC) resulted in 80% protection against GPS5 infection in mice, along with significantly increased antigen-specific IgG levels compared to controls. In conclusion, MEVC is a promising vaccine candidate to protect against porcine Glasser's disease.