DDA/TDB adjuvant significantly increased BCG-A4-mediated IFN-γ secretion from pulmonary DN T cells. Mice were treated with antibodies to deplete CD4+, CD8+, and NK1.1+ T cells before receiving an aerosol challenge with M. tuberculosis Erdman. Lung cells were isolated from nonvaccinated mice or mice vaccinated with the BCGΔmmaA4 mutant (BCG-A4) or A4/Adj 10 days postchallenge. The frequency and MFI values of pulmonary DN T cells secreting IFN-γ were measured by flow cytometry. The iMFI is the integrated median fluorescence intensity, which is calculated by multiplying the frequency of cells producing IFN-γ by the MFI value. Means and SD were calculated from 3 mice per group from one experiment. *, significant differences relative to nonvaccinated mice (P < 0.05). #, significant differences relative to BCG-A4-immunized mice (P < 0.05).