Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease (JD) and has a major economic impact on the livestock industry in the United States. Progress in developing an effective vaccine against JD is impeded by the lack of knowledge of induction, persistence and dysregulation of protective immunity. We have proposed to develop a membrane vesicles and E. coli outer membrane vesicles delivery system for use in a vaccination program against Johne’s disease. Outer membrane vesicles recently have proven successful as vaccine delivery systems for human use. In the proposed research we plan to evaluate the short term immune response against membrane vesicles and E. coli outer membrane vesicles in goats. To accomplish these objectives, we will analyze immune responses in blood and tissues of vaccinated and challenged animals by employing various modern techniques including RT-PCR, FACS, and ELISA/ELISPOT analysis. The results obtained will be utilized to better understand MAP pathogenesis and immune evasion. Our long-term goal is to develop a prophylactic intervention strategy against MAP. Our short term goal is to analyze various immune cells and markers associated with protective immunity. The proposed research aims to increase our knowledge and understanding of the protection mechanism after vaccination with membrane vesicles and E. coli outer membrane vesicles in a goat model, which may lead to development of effective strategies to control JD. The Program Area Priority we are addressing is Disease prevention and control (e.g., vaccines, diagnostics, enhanced innate or adaptive immunity, disease resistance or susceptibility, or management)