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. 2020 Oct 9;70(6):1023–1036. doi: 10.1136/gutjnl-2020-321731

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© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See: https://creativecommons.org/licenses/by/4.0/.

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IL-1α and IL-1β signalling are essential for monocyte IL-23 production. PBMC from healthy donors (n>4) were stimulated for 16 hours with combinations of LPS and aIL-10R in the presence of indicated exogenous human recombinant cytokine (all 10 ng/mL) and/or cytokine/cytokine receptor blockade (all antibodies 10 µg/mL). (A) Frequencies of IL-12p40⁺IL-23p19⁺ live CD14⁺ monocytes (Wilcoxon test, 95% CI). (B) Representative dot plot showing intracellular IL-12p40 and IL-23p19 according to (A). (C) tSNE presentation of IL-23p19, CCL20, HLA-DR, IDO-1, CCL2, S100A8, RPS6 and SPINK-1 expression in live CD14⁺CD3^–CD19^–CD56^–-gated monocytes. Analyses of three healthy donors are shown as overlay. (D) Frequencies of monocyte clusters across stimulations based on cluster-specifying protein expression (n=6; one-way analysis of variance after BH correction). (E) The extensive model represents the effects of the addition or blockade of cytokines in a PBMC culture in the presence of LPS or LPS and anti-IL-10R. Differences of cytokine addition or blockade in LPS-stimulated samples (dashed arrows), in LPS and anti-IL-10R-stimulated samples (solid arrows) and in LPS and anti-IL-10R-stimulated and anti- IL-1β treated conditions (dotted arrows). Nominal effects with p>0.05 after false discovery rate correction (BH) are shown in parentheses. (F) A reduced complexity model was established by focusing on informative cytokine interactions. Edge weights are defined as the relative contribution to model fit and are dependent on the network configuration considered. The edges of the 20-edge model have been coloured based on their weight. BH, Benjamini-Hochberg; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; PBMC, peripheral blood mononuclear cells; TNF, tumour necrosis factor.