FIGURE 8.

rMgTx decreases the expression of IL2R (CD25) and CD40L. Isolated CD4⁺ T_EM cells were stimulated with anti-human CD3 antibody in the presence or absence of toxins. After 24 h of stimulation, cells were labelled with anti-CD25 (IL2R) (A,B) and anti-CD154 (CD40L) antibodies (C,D). Treatment labels: US, unstimulated (black); S, stimulated with anti-human CD3 antibody–coated wells (5 µg/well; see the Materials and Methods section for details, green); S + TrMgTx, stimulated in the presence of TrMgTx (8.5 nM, red); S + UrMgTx, stimulated in the presence of UrMgTx (5 nM, blue). (A,C) Fluorescence histograms were obtained from T lymphocytes gated based on their FSC and SSC parameters (10,000 events were recorded), and then, histograms corresponding to CD25 or CD154 expression were plotted as peak-normalized overlay. Plots were made using FCS Express 6.0. (A,C) has the same color code. (B,D) Mean fluorescence intensities (MFI) were determined from the histograms and normalized to that of their stimulated but not treated control (S). Data represent values from three independent experiments (two technical repeats in each) with SEM. Significant differences of IL2R and CD40L expression between the stimulated samples in the absence and presence of toxin is indicated with asterisks (*p < 0.05, **p < 0.01, all pairwise multiple comparison).