Conjugated Bilirubin Differentially Regulates CD4+ T Effector Cells and T Regulatory Cell Function through Outside-In and Inside-Out Mechanisms: The Effects of HAV Cell Surface Receptor and Intracellular Signaling

Figure 4

Treatment with CB in vitro induces an augment in the activity of Tregs. Purified CD4+CD25+FOXP3+ Tregs from pediatric healthy donors were incubated for 72 hours with and without different doses of CB. Then, Tregs were cocultured with CFSE-labeled Teff (TCD4+CD25−) for 7 days, and cells were stimulated with anti-CD3 and anti-CD28. (a) Dilution of fluorescence and cell count because of the proliferation of Teff stimulated with anti-CD3 and anti-CD28 cocultured with Tregs in the absence of CB. (b) Dilution fluorescence and cell count because of the proliferation of Teff when cocultured with Tregs with CB 0.3 mg/dL. (c) Dilution fluorescence and cell count because of the proliferation of Teff when cocultured with Tregs preincubated with CB 2 mg/dL. Data shown are representative of three independent experiments. (d) The results are displayed as the percentage of Tregs suppression by each condition. (e) IL-17 concentration was evaluated by MAGPIX Technology in cell supernatants from CD4+ TLs from pediatric healthy donors stimulated with anti-CD3 and anti-CD28 in presence or absence of CB. The medians and standard deviations of three repetitions are presented. Nonparametric Kruskal-Wallis for comparison between groups was used to calculate statistical significance. was considered statically significant. .

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