Paratuberculosis is a chronic infectious disorder and a major problem in farmed ruminants. This disease is caused by Mycobacterium avium subsp. paratuberculosis. M. avium subsp. paratuberculosis is an important pathogen that causes Johne's disease in animals and also has been implicated as a possible cause of Crohn's disease in humans, but little is known about the protective immune responses to this microorganism. Fibronectin attachment protein (FAP) is a member of a family of fibronectin-binding proteins produced by several species of mycobacteria which is important in the pathogenesis of M. avium. Addition of recombinant FAP to human respiratory tract organ cultures inhibits M. avium binding to areas where there is epithelial damage. We characterized the role of FAP in promoting adaptive and innate immune responses. FAP functionally activated dendritic cells by augmenting the expression of CD80, CD86, major histocompatibility complex class I, and major histocompatibility complex class II. Moreover, FAP induced the allogeneic immunostimulatory capacity of dendritic cells by stimulating dendritic cell production of Th1-promoting interleukin-12. FAP also increased the production of gamma interferon by T cells in mixed-lymphocyte reactions, which would be expected to contribute to the Th1 polarization of the immune response. The expression of surface markers and cytokine production in dendritic cells was mediated by both mitogen-activated protein kinases and NF-kappaB pathways. These results show that FAP modulates the adaptive immune responses to M. avium subsp. paratuberculosis by inducing maturation and activation of dendritic cells, which drives Th1 polarization.