Fig. 2 | Nature Communications

Fig. 2

From: The cholesterol biosynthesis pathway regulates IL-10 expression in human Th1 cells

Fig. 2

T-helper type 1 (Th1) switching to interleukin-10 (IL-10) is blocked when the mevalonate pathway is inhibited. Purified human CD4+ T cells stimulated in vitro with plate-bound α-CD3 (2 μgml−1) + α-CD46 (5 μgml−1) and recombinant human interleukin-2 (rhIL-2) (50 Uml−1) were cultured for 36 h in the presence of atorvastatin and 250 μM mevalonic acid (MA) as indicated, unless stated otherwise. a Representative flow cytometric analysis of intracellular interferon-γ (IFNγ) and IL-10 staining. b Normalised frequency of IFNγIL-10+- (left), IFNγ+IL-10+- (centre) and IFNγ+IL-10- (right) producing cells (n = 20). c Normalised concentration of secreted IL-10 (n = 19). d Normalised IL10 messenger RNA (mRNA) levels (n = 8). e Representative flow cytometric analysis of intracellular IL-17 (top) and tumour necrosis factor-α (TNFα) (bottom) co-stained with IL-10 of atorvastatin-treated cells (cumulative data shown in supplementary Fig. 3g). f Normalised concentration of secreted IL-4 (n = 9). g Effect of atorvastatin (AT) treatment on purified CD4+ cells co-cultured with monocytes and α-CD3 (100 ng ml−1) in the absence or presence of Adalimumab (1 μgml−1). Data show a representative dot-plot for intracellular IL-10 and IFNγ production (cumulative data shown in Supplementary Fig. 3h). Graphs show independent donors (dots) normalised to 0 μM dose of atorvastatin; bars represent median values. *<0.05, **<0.01 and ***<0.001 denote a significant difference compared to untreated cells by repeated-measures one-way analysis of variance (ANOVA) test with post hoc Dunnett’s correction (b, d, f) or Friedman test with post hoc Dunn’s correction (c)

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