Extended Data Fig. 5: Immune characterization of circulating monocytes.

(a) Gating strategy used for identification of circulating monocytes in the peripheral blood. Live CD45⁺ cells were gated to remove CD3⁺ T cells (CD3-) and CD11b⁺Ly6G⁺ neutrophils (neutrophil-). The remaining cell population was gated to identify classical CD11b⁺Ly6C^high and nonclassical CD11b⁺Ly6C^low circulating monocytes. (b) Representative histograms (n = 3 mice/group) comparing levels of MHCII expression on Ly6C^high and Ly6C^lowmonocytes in the peripheral blood of BCG-vaccinated and PBS-treated hosts cultured with (S) and without stimulation (US). (c & d) Frequencies of IL-6-producing Ly6C^high and Ly6C^low monocytes and frequencies of TNF-producing Ly6C^high monocytes with (S) and without (US) stimulation. n = 3 mice/group. (e) IL-6 (*p = 0.015), IL-1β (***p = 0.0007), TNF, IL12p40 (*p = 0.0173), IP10 (*p = 0.0229), MIP-1-α (*p = 0.0407) and RANTES (**p = 0.0024) protein contents in the plasma (pg/ml) of BCG-vaccinated or PBS control animal-derived whole blood samples cultured with (S) and without (US) stimulation. n = 3 mice/group. Data are presented as mean ± SEM, representative of two independent experiments. Statistical analysis was determined by two-tailed t test for data in e.