IL-10 regulates IL23A transcription and IL-23 protein secretion in a subset of monocytes from patients with IBD. (A) Analysis of PBMC culture supernatants collected after 16 hours of stimulation with LPS±IL-10R blocking antibodies expressed as log2 fold change to unstimulated PBMC (Ctrl) or LPS-stimulated PBMC (n=28; mean±SEM; Wilcoxon test, BH-adjusted p values). (B) IL-23 protein concentrations in culture supernatants expressed as fold change to unstimulated PBMC (n=28; mean±SEM; Friedman test with FDR-adjusted p values). (C) RT-qPCR analysis of relative IL23A expression in PBMC following 16 hours of stimulation (n=45; mean±SEM; Kruskal-Wallis test, BH-adjusted p values). (D) Contour plot presentation of IL-23p19-producing, IL-12p40-producing and IL-10-producing live leukocytes and CD14 surface expression measured at 16 hours poststimulation in PBMC. (E) Frequencies of IL-12p40+IL-23p19+, IL-10+ and IL-12p40+IL-23p19– CD14+ and CD14– cells in total live leukocytes (n=18, mean±SEM, Mann-Whitney test). (F) Frequencies of IL-12p40+IL-23p19+, IL-10+ and IL-12p40+IL-23p19– of CD14+ cells (n=18, mean±SEM, Mann-Whitney test). BH, Benjamini-Hochberg; Ctrl, control; EGF, epidermal growth factor; FDR, false discovery rate; G-CSF, granulocyte-colony stimulating factor; IL, interleukin; LPS, lipopolysaccharide; MDC, macrophage-derived chemokine (CCL22); n.s., not significant; PBMC, peripheral blood mononuclear cells; RANTES, regulated upon activation - normal T cell expressed and presumably secreted (CCL5); RT-qPCR, real-time quantitative PCR; TNF, tumour necrosis factor. *pvalue 0.05, **pvalue<0.01, ***pvalue<0.001, ****pvalue<0.0001.