Figure 7.

Feed-forward action of induced cytokines amplifies innate response to bacillus Calmette–Guérin (BCG). (A–C) The concentrations of cytokines were measured for whole blood and purified monocytes from healthy donors stimulated for 5 h with BCG in the presence of recombinant IL-1RA, anti-TNFR, and anti-IL-6R antibodies (referred to as “all blockers,” n = 6). (A) Detailed results from whole blood were represented as pair analyses, each line indicates a donor and the thick black line represents median values. Heat maps’ representation of the median concentration for the listed cytokines for whole blood [(B), n = 6] and purified monocytes [(C), n = 4–8], stimulated with BCG in the presence of the indicated blocking receptor reagent(s), BCG in the presence of isotype control antibodies, or buffer control [non-stimulated (NS)]. Percent inhibition between “all blockers” condition and BCG condition is indicated for each cytokine. (D–G) Purified monocytes (n = 7), neutrophils (n = 9), or lymphocytes (n = 7) were stimulated by BCG in the presence of increasing doses of the JAK1/2 inhibitor Ruxolitinib, and the quantification of cytokines were measured using Luminex assay. (D) Box-whisker plots indicate the percentage inhibition as compared to BCG stimulation for purified monocytes (normalized to 100% across experiments). The dotted lines indicate the median values for BCG stimulation for each cytokine. (E) Heat maps represent the median concentration determined for monocytes, (F) neutrophils, and (G) lymphocytes. p Values were determined by the paired Student’s t-test and false discovery rate corrected for multiple analyte testing. *q ≤ 0.05; **q ≤ 0.01; ***q ≤ 0.001 as compared to BCG + isotype control or DMSO control.