Figure 5.

CB in vitro induces an augment in the percentage of Tregs expressing TIM-1. Purified Tregs from pediatric healthy donors were incubated at 37°C, 5% CO₂ with different concentrations of CB for 72 h. Then, cells were recovered and stained with anti-TIM-1-PE and the percentage of Tregs expressing TIM-1 was analyzed by using flow cytometry. Representative dot plots are shown. (a) Tregs without stimulus. (b) Tregs stimulated with anti-CD3 and anti-CD28. (c) Tregs stimulated with anti-CD3, anti-CD28 and CB (0.3 mg/dL). (d) Tregs stimulated with anti-CD3, anti-CD28, and CB (2 mg/dL). (e) Tregs stimulated with anti-CD3, anti-CD28, and CB (15 mg/dL). (f) The results are presented as TIM-1 percentage. The medians and SD of three independent experiments are shown. Nonparametric Kruskal-Wallis for comparison between groups was used to calculate statistical significance. ^∗ P < 0.05 was considered statistically significant. ^∗∗ P < 0.001.