Live Mycobacterium avium subsp. paratuberculosis and a Killed-Bacterium Vaccine Induce Distinct Subcutaneous Granulomas, with Unique Cellular and Cytokine Profiles

Abstract

ABSTRACT Type II (lepromatous) granulomas are characterized by a lack of organization, with large numbers of macrophages heavily burdened with bacilli and disorganized lymphocyte infiltrations. Type II granulomas are a characteristic feature of the enteric lesions that develop during clinical Mycobacterium avium subsp. paratuberculosis infection in the bovine. Considering the poor organization and function of these granulomas, it is our hypothesis that dendritic cell (DC) function within the granuloma is impaired during initial infection. In order to test our hypothesis, we used a subcutaneous M. avium subsp. paratuberculosis infection model to examine early DC function within M. avium subsp. paratuberculosis -induced granulomas. In this model, we first characterized the morphology, cellular composition, and cytokine profiles of subcutaneous granulomas that develop 7 days after subcutaneous inoculation with either vaccine or live M. avium subsp. paratuberculosis . Second, we isolated CD11c + cells from within granulomas and measured their maturation status and ability to induce T-cell responses. Our results demonstrate that M. avium subsp. paratuberculosis or vaccine administration resulted in the formation of distinct granulomas with unique cellular and cytokine profiles. These distinct profiles corresponded to significant differences in the phenotypes and functional responses of DCs from within the granulomas. Specifically, the DCs from the M. avium subsp. paratuberculosis -induced granulomas had lower levels of expression of costimulatory and chemokine receptors, suggesting limited maturation. This DC phenotype was associated with weaker induction of T-cell proliferation. Taken together, these findings suggest that M. avium subsp. paratuberculosis infection in vivo influences DC function, which may shape the developing granuloma and initial local protection.