Oral immunization with recombinant Saccharomyces cerevisiae expressing TM1 of Mycoplasma gallisepticum induces unique specific antibodies and protective immunity

Mycoplasma gallisepticum (MG) is one of the main pathogens causing chronic respiratory diseases in chickens, which seriously affects the sustainable and healthy development of the poultry industry and leading to heavy economic losses. Therefore, we developed a safe, efficient, convenient, and low-cost MG oral vaccine. The vaccine is based on a recombinant yeast surface display system to compensate for the shortcomings of existing vaccines. TM1, a protective antigen protein, is located on the outer membrane of MG cells. In this study, the physical and chemical properties, structure and epitopes of MG protective antigen protein were systematically analyzed by bioinformatics method, and TM1 protein was predicted to be suitable for vaccine development as MG immunogenic protein. Subsequently, we constructed recombinant S. cerevisiae expressing MG-TM1 protein and evaluated its immunoprotective effects following oral administration. First, the recombinant S. cerevisiae strain EBY100/pYD1-TM1 efficiently expressed TM1, as confirmed by high-throughput screening, PCR, and Western blot analysis. After oral administration with EBY100/pYD1-TM1 stimulates the high level of IgG and sIgA in chicken. Notably, it prevents MG colonization and effectively protected the respiratory system such as air sacs and lungs from damage. The above results suggest that EBY100/pYD1-TM1 can induce humoral and mucosal immune responses in chicken. Collectively, the recombinant S. cerevisiae expressing TM1 is a potentially safer and more effective agent against MG infection.