Fig. 1: Subcutaneous BCG vaccine induces memory AM in distal respiratory mucosa.

a, Experimental schema. b,c, Histograms of surface expression of MHC II (**P = 0.0027, ****P < 0.0001) (b) or TLR2 (**P = 0.0050) (c) on airway-resident AM. d,e, Histograms and frequencies of IL-6- (***P = 0.0007) (d) or TNF-producing airway AM (e). f, Heatmap of cytokine/chemokine protein levels (geometric means) in supernatants of airway AM cultured with (S) or without (US) stimulation. Red asterisks denote significantly increased cytokine/chemokine production upon stimulation by airway AM of BCG hosts. g–l, Concentrations of IL-6 (*P = 0.0230) (g), IL-12p40 (****P < 0.0001) (h), TNF (i) and MCP-1 (**P = 0.0031) (j), MIP-1α (*P = 0.0408) (k) and RANTES (*P = 0.0137) (l) in supernatants of airway AM cultured with and without stimulation. m, Real-time ECAR in airway AM at 8 weeks post-BCG immunization. 2-DG: 2-deoxy-glucose. n, Glycolysis (***P = 0.0006), glycolytic capacity (***P = 0.0007) and glycolytic reserve (**P = 0.0013) in airway AM at 8 weeks post-BCG immunization. o, Dotplots of frequencies of cells gated out of total live peritoneal cells expressing F4/80 surface marker and the median fluorescent intensity (MFI) of F4/80 expression by PM at 5 weeks post-BCG vaccination (*P = 0.0231, ****P = <0001). p, Histograms of and the MFI of MHC II and frequencies of IL-6- or TNF-producing PM at 5 weeks postvaccination with viable BCG or inactivated BCG (BCG-ia) or PBS with (S) or without (US) stimulation (****P = <0.0001). Data in b, d and e are representative of three independent experiments. Data in f, g–l and o–p are representative of two independent experiments. Data in bar graphs are presented as mean ± s.e.m. and represent individual data points of biologically independent samples, n = 3 mice per group. One-way ANOVA was used for multiple comparison testing with Bonferroni’s test for data in b–l, two-tailed t-test for data in n and one-way ANOVA followed by Dunnett’s multiple comparison test for data in o and p.