Extended Data Fig. 1: Characterization of innate memory phenotype of alveolar macrophages following s.c. BCG vaccination.

(a) MFI of MHC II expression on lung tissue AM with (S) or without (US) stimulation (**p = 0.0012, ***p = 0.0004). (b & c) Frequencies of IL-6- and TNF-producing lung tissue AM with (S) or without (US) stimulation. (d) Real-time oxygen consumption rate (OCR) in airway AM at 8-wks post-BCG vaccination. (e) Experimental schema. (f & g) Histograms of incorporation of BrdU by airway AM and frequencies of BrdU-incorporated airway AM (*p = 0.0475) (f) & measure of MFI of MHCII on BrdU + (*p = 0.0430) and BrdU- (p = 0.0519) airway AM (g). n = 3 mice/group. (h) Experimental schema. (i, j & k) Measure of MFI of MHCII on airway and lung tissue AM (*p = 0.0365) (i), frequencies of IL-6- (j) and TNF- (k) producing airway and lung tissue AM (*p = 0.0249). n = 3 mice/group/airway, 4 mice/group/lung tissue. (l) Real-time extracellular acidification rate (ECAR) in airway AM and glycolysis, glycolytic capacity and glycolytic reserve in airway AM (****p = 4 mice/group. (m) Measure of MFI of MHCII expression on airway AM (*p = 0.0497) and frequencies of IL-6- (**p = 0.0082) and TNF-producing airway AM at 5-wks post-vaccination with viable BCG or inactivated BCG (BCG-ia) or PBS with (S) or without (US) re-stimulation. n = 3 mice/group. Data in a-c are representative of two independent experiments (n = 3 mice/group). Data in bar graphs are presented as mean ± SEM. One-way ANOVA was used for multiple comparison followed by Bonferroni’s test for data in a-c and i-k, two-tailed t test for data in f, g & l and one-way ANOVA, followed by Dunnett’s multiple comparison test for data in m.