Figure 2
From: A novel lipidic peptide with potential to promote balanced effector-regulatory T cell responses
IK14004 suppresses DC licensing and maturation. Immature monocyte-derived dendritic cells (iMoDCs) were prepared as described in the “Methods”. Each tissue culture experiment involving use of either monocytes, PBMCs or isolated CD3+ T cells was performed using triplicate wells (technical replicates) and flow cytometry/ELISA experiments were repeated at least three times (n = experimental replicates) as indicated below each panel. All error bars represent standard error of the mean (SEM). In all experiments the culture duration was 72 h. The compound shown is IK14004 as indicated under each panel. Flow cytometry data are shown as mean fluorescence intensity (MFI). Dot plots and gating strategies are shown in Supplementary Figs. S10 to S17. (a) Expression of CD40L in CD4+ T cells within T cell cultures. (b) Expression of CD40L in CD4+ T cells within PBMC cultures. (c) Expression of CD40L in CD8+ T cells within T cell cultures. (d) Expression of CD40L in CD8+ T cells within PBMC cultures. (e) Viability of immature monocyte-derived dendritic cell (iMoDC) cultures. (f) Expression of CD86 in iMoDC cultures. (g) Expression of CD25 in iMoDC cultures. (h) IL-12p40 levels in supernatant from iMoDC cultures. (i) IL-12p40 production by iMoDCs expressed as fold-change between the highest and lowest IK14004 concentration, respectively, with vehicle-treated cells normalised to 1. (j) Reversion of CD14 negative iMoDCs to CD14+ monocytes. Data were analysed using repeated measures (RM) two-way ANOVA with Dunnett’s post-test comparing peptide with vehicle control. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
