Abstract: Staphylococcus aureus (SA), one of the major causes of livestock infection, is resistant to many kinds of antimicrobials. It is urgent to develop vaccines targeting at this pathogen. In this study, a fusion protein (ABF) composed of EsxA, EsxB and Flic was firstly generated. To examine the effect of ABF against SA, Banna mini-pig inbred line wound models were established and divided into four groups, the control wound group, the SA infected group, the EsxB pre-immunized group and the ABF pre-munized group. The results showed that peripheral IgG expression level in the mini-pig pre-immunized with ABF was significantly higher than that of the other three groups. Besides, ABF could activate T cell as well. The ratios of CD8a, CD3e and IFN-γ positive cells to spleen lymphocytes were all increased in the ABF group. In conclusion, all the results showed that ABF could significantly induce chemokine response and was functional during wound healing.