Fig. 4

ATT impairs the innate immune response. Mice were treated with RIF or INH/PYZ for 8 wks. Treatment was stopped 2 days prior to Mtb. After 5d post infection, single-cell suspensions of lung cells were immunophenotyped by flow cytometry. a Frequency and b total cell number of pulmonary monocytes, interstitial macrophages, alveolar macrophages, eosinophils, and neutrophils as determined by the lineage-specific markers described on the y axis of each graph. c, d Expression of MHCII on c alveolar macrophages and d monocytes and interstitial macrophages. e–g Alveolar macrophages were harvested from INH/PYZ-treated mice and, using a Seahorse analyser, mitochondrial function was analysed through the addition of mitochondrial inhibitors in the following order: oligomycin A (1.5 µm), carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP) (2 µm), and antimycin A (2 µm) as a function of time. The oxygen consumption (OCR), basal respiration, maximal respiration, spare respiratory capacity and ATP production was quantified in f and extracellular acidification rate (ECAR) was measured as shown in g. h, i Alveolar macrophages were harvested at 8 weeks post treatment and infected with Mtb in vitro. 12 h post infection mRNA expression and secretion of h TNF-alpha and i IL-1β as determined by qRT-PCR and ELISA, respectively. j, k Alveolar macrophages were harvested from naive mice and incubated with RIF or INH/PYZ for 2 h. Cells were washed, rested overnight, and the metabolic capacity was assessed as described for e–g. Data shown are representative of two independent experiments. Error bars represent mean ± SEM; *p < 0.05, ***p < 0.001, ****p < 0.0001; nd, not detected