The research including preparation an antigens of killed whole cell sonicated antigen of Salmonella typhimurium (KWCSAg-ST) and killed whole cell sonicated antigen of Staphylococcus aureus (KWCSAg-STAPH.A), then evaluated their efficacy by using delayed type hypersensitivity (DTH)-skin test, and enzyme linked immuno-sorbent assay (ELISA). Twenty four rabbits randomly divided into three groups (eight animals for each group). The first group were immunized with KWCSAg-ST (1mg/ml) subcutaneously. The second group was immunized with KWCSAg-ST (1mg/ml) and KWCSAg-STAPH.A (1mg/ml) subcutaneously. The third group was injected by PBS (pH7.2) as control group. The highest diameter of the erythema was 22.00±1.67mm for 1st group as compared with 2nd group 16.00±0.58mm. There was an increase in the diameter of induration in 1st group at 48 hours. The highest diameter as recorded 15.33±1.52mm of 1st group compared with a significant difference (P<0.05). The immune response was estimated by using ELISA, The value of IgG concentration in 3rd group showed the elevation to 110.57±7.48µg at days 28 post immunization then decrease gradually to reach 92.29±4.22µg at days 56 post immunization with significant difference (P<0.05). IgG concentration in 1st and 2nd groups was 107.21±4.61µg, and 108.83±5.97µg, at 28 post immunized respectively, with significant different (P<0.05), then decrease gradually to reach 108.83±10.63µg and 100.62±5.53µg at days 56 post immunized respectively with significant difference (P<0.05). While The IL-4 concentration in 3rd groups elevated to 25.11±1.88pg at days 28 post immunization then decrease gradually to reach 23.74±2.93pg at days 56 post immunization with significant difference (P<0.05). IL-4 concentration in 1st and 2nd groups was 23.94±0.83pg and 25.56±1.90pg and at day 28 post immunized respectively, and elevated gradually to reach 27.28±3.59pg and 26.89±2.37pg at day 56 post immunized respectively with significant difference (P<0.05). In conclusion there was an interaction of the two antigens to stimulate and progress the cellular and humoral immune response, likewise give immunological reaction between bacterial antigens of Salmonella typhimurium and Staphylococcus aureus which act as adjuvant to give good potent synergistic effect on immunological response.