Antigen-specific proliferation and activation of peripheral blood mononuclear cells from Mycobacterium bovis-infected...

To evaluate antigen-specific proliferative and activation-associated responses from Mycobacterium bovis-infected reindeer, blood mononuclear cells from M. bovis- (n = 10) and non-infected reindeer (n = 4) were stimulated with a recombinant early secretory antigenic target-6 and culture filtrate protein-10 fusion protein (rESAT6:CFP10), M. bovis purified protein derivative, pokeweed mitogen, or medium alone and evaluated by flow cytometry using dye tracker analysis and cell surface marker staining. gammadelta TCR+ and CD8+ cells, but not CD4+ cells, from M. bovis-infected reindeer proliferated in response to specific antigen stimulation. Expression (i.e., mean fluorescence intensity) of CD44 was increased and CD62L decreased on proliferative as compared to non-proliferative fractions in antigen- and mitogen-stimulated cultures. In response rESAT6:CFP10 stimulation, MHC II fluorescence intensity was increased on CD4+, gammadelta TCR+, CD172a+, and IgM+ cells from infected reindeer as compared to that of non-stimulated cells from the same reindeer. Recombinant ESAT6:CFP10 stimulation also induced expansion of a CD172a+, MHC II+ population within mononuclear cell cultures from M. bovis-infected reindeer. Despite a moderate challenge dose and extended duration of incubation, experimental infection of reindeer was generally limited to lymph nodes draining the inoculation site, suggestive of host resistance to progressive disease. Present in vitro findings, therefore, may be predictive of host responses by reindeer that limit progression to disseminated disease.