Developing an effective vaccine against Staphylococcus aureus (S. aureus) is a key global health concern, especially with the increased reports of multidrug-resistant (MDR) S. aureus strains. Previous attempts for S. aureus vaccine development were unsuccessful. In this study, Manganese transport protein C (MABC) B cell epitopes, Nickel ABC transporter (NABC) B cell & T cell epitopes, and Phosphatidylinositol phosphodiesterase (PIc) B cell & T cell epitopes were used as a vaccine in mice skin infection model. Mice immunized with peptide mixture and MABC peptide group showed the best skin lesion healing results. The protection level was correlated with the highest IgG level, highest levels of interferon-gamma (INF γ), and lowest levels of interleukin-2 (IL-2). The peptide mixture group also showed the highest count of CD4/ CD8 cells. Results demonstrated that the inclusion of B cell and T cell epitopes of multiple genes improved both the humoral and cellular immunity and resulted in the best outcome in the skin infection mice model. A more expanded in-vivo study in different mice models is recommended for testing MABC, NABC, and PIc B cells and T cells peptides cocktail as promising S. aureus vaccine.