Mycoplasma bovis is a primary infectious agent of many disorders in cattle including bovine respiratory disease. No commercial vaccines against M. bovis are available in Europe. The immune response of calves to three saponin-based adjuvants combined with a field Polish M. bovis strain was evaluated. Four groups of six calves each were injected subcutaneously with the M. bovis strain combined with either saponin, saponin + Emulsigen®, saponin + Emulsigen® + alphatocopherol acetate, or with phosphate-buffered saline as control group. Blood and nasal swab samples were collected up to day 84 post injection. All formulations effectively stimulated the humoral and the cellular immune response of the calves, but the course of the response depended on the adjuvant formulation. These immunological data provide additional information supporting the findings of previous M. bovis saponin and Emulsigen® vaccine challenge studies to facilitate the development of successful M. bovis vaccines.