Figure 7.

Adjunct respiratory mucosal (RM) immunotherapy enhances antigen (Ag)-specific CD8 T cell responses in the lung and confers protection in a T cell-dependent manner. (A) Experimental schema. Mice were set up as depicted in experimental schema Figure 3A. (B) Representative dot plots depicting the frequencies and bar graph depicting absolute numbers of Ag85A-specific CD8 T cells as assessed by tetramer immunostaining. (C) Representative dot plots depicting the frequencies and bar graph depicting absolute numbers of Ag85A-specific IFNγ ⁺ T cells as assessed by intracellular cytokine staining after recombinant Ag85A stimulation of lung mononuclear cells. (D) Experimental schema. Mice were infected and treated with antibiotics as per Figure 1A. One set of animals received RM immunotherapy 4 weeks after the initiation of antibiotics (ABx/Vac.). T cells were subsequently depleted weekly after immunotherapy (ABx/Vac. T cell depleted). A set of animals received a single dose of AdCh68 empty control vector in place of AdCh68Ag85A (ABx/empty vector). Data are expressed as the mean ± standard error of the mean of 3 mice/group for B or C and 6–10 mice/group for D. ABx, antibiotics alone; CFU, colony-forming units.