4.0 (San Diego, CA). The statistical significance of differences between two groups was tested using a Student’s t-test. For comparison of more than two groups, Kruskal–Wallis one-way analysis of variance (anova) was used. If the anova was significant, the Tukey–Kramer test was used as a post hoc test. Differences of P < 0·05 were considered significant. All data are expressed as means ± SEM, *P < 0·05, **P < 0·01, ***P < 0·001. Conventional immature DCs were generated from monocytes by 6 days of culture with GM-CSF and IL-4. Other stimuli were added during the differentiation process;

TCDCA (100 μm) for TCDCA-DCs, TGR5 agonist (20 μm) for TGR5-DCs, 8-Br-cAMP (10 μm) for cAMP-DCs, and fexaramine (100 μm) for FXR-DC. These DCs revealed Dorsomorphin different morphology and cell surface antigen click here expression (Fig. 1a,b). We observed BA-DCs, TGR5-DCs and FXR-DC expressing low levels of CD1a, but not cAMP-DCs. Expression of co-stimulatory molecules, CD80 and CD86, was increased in BA-DCs, TGR5-DCs, cAMP-DCs and FXR-DCs. These findings demonstrated

that TCDCA, TGR5 agonist, cAMP and FXR agonist induce different types of DCs during the 6-day differentiation culture. The viability of cDC, TCDCA-DCs, and TGR5-DCs was also confirmed (see Supplementary material, Fig. S1). We have previously found that retinoic acid affects the differentiation of DCs from monocytes and induces anti-inflammatory DC differentiation.7 We hypothesized Farnesyltransferase that BAs might also affect the differentiation of DCs. To assess this, we cultured DCs differentiated from monocytes

in the presence (referred to as BA-DCs) or absence (referred to as cDCs) of a BA and measured the cytokine-producing ability of these cells following stimulation with heat-killed antigen from the commensal bacteria E. faecalis or LPS + interferon-γ. The BA-DCs produced significantly less of the pro-inflammatory cytokines IL-12p70 and TNF-α in response to bacterial antigen or LPS + interferon-γ stimulation than cDCs, in a manner that was dependent on the concentration of the BA (Fig. 2a,b). We next investigated whether the FXR signalling pathway was involved in the DC differentiation process, using fexaramine, a powerful synthetic FXR agonist, in place of the BA during DC differentiation from monocytes. Unexpectedly, DCs differentiated in the presence of the FXR agonist did not show the same IL-12 hypo-producing DC phenotype as DCs differentiated in the presence of the BA (Fig. 3a,b). We also examined mRNA expression of BA transporters, bile salt export pump (BSEP), organic anion transporting polypeptide C (OATP), sodium taurocholate cotransporting polypeptide (NTCP) and apical sodium-dependent bile salt transporter (ASBT) on monocytes and DCs. As shown in Fig. 3(c), no transporters for BAs were expressed on peripheral blood monocytes. The transporter BSEP was expressed in DCs, but all other transporters were absent in both monocytes and DCs.