Fig. 5

Effect of tanimilast on CD141 (Thrombomodulin/BDCA3) expression in DCs. A, B moDCs were pre-treated or not (-) with tanimilast (TAN) or budesonide (BUD) (both at 10⁻⁷ M) for 1 h and subsequently stimulated with LPS for 24 h. The expression of CD141 was evaluated by FACS analysis (A) and by Real time PCR (B). Data are expressed as mean ± SEM (n = 3) of the percentage of positive cells (left y axis) and of the Median Fluorescence Intensity (MFI) (right y axis) (A) and of 2^−ΔΔCt relative to (-) (fold induction at 24-h time-point) (B). moDCs (C) or CD1c⁺ DCs (D) were treated as described in A and subsequently stimulated or not with E. coli, R848, β-glucan (C) or LPS (D) for 24 h. The surface expression of CD141 were evaluated by FACS analysis. Data are expressed as the mean ± SEM (n = 3) of the percentage of positive cells (left y axis) and of the Median Fluorescence Intensity (MFI) (right y axis). ^#P < 0.05 versus (-) and *P < 0.05 versus LPS (A, B, D) or E. coli, R848, β-glucan (C) by one-way ANOVA with Dunnett’s post-hoc test. E moDCs treated as described in A were labeled with MHC-II and then co-cultured with heat-killed autologous CFSE⁺ PBMC at ratio 1:1 for 2 h. Dead cells uptake was defined as the percentage of MHC-II⁺CFSE⁺ cells. Left panel represents the fold increase of dead cell uptake upon LPS stimulation. Data are expressed as the mean ± SEM (n = 3) of the fold increase. Right panel represents the correlation between the percentage of CD141⁺ cells and dead cell uptake by LPS-moDCs (black dots), TAN-LPS-moDCs (dark grey dots) and BUD-LPS-moDCs (light grey dots) from 3 donors (R² = 0.82; Spearman r = 0.9055, ***P = 0.0008)