Fig. 2
In vitro functional characterization of sTreg cells. a T-cell suppression (CFSE dilution) assay. DCs (1 × 105 cells) of FVB/J mice and 1 × 105 CFSE-labeled CD4+CD25− T cells (HA mice) were added to set up T-cell suppression assay. The nTreg or sTreg cells were added in each well at different ratios of T cells and cultured for 5 days. The proliferation of T cells in mixed lymphocyte reaction was measured in terms of dilution of CFSE intensity as shown in the representative histograms. b A significant (*P = 0.0142) difference in suppression of T-cell proliferation between two groups of Treg cells was noticed at 0.1:1, 0.25:1, 0.5:1, and 1:1 ratios of the regulatory to effector T cells. c IL-10 secretion in the culture supernatants (Treg cells: T cells = 1:1) was measured by enzyme-linked immunosorbent assay. A significant increase in IL-10 levels was observed in cultures with sTreg cells (P = 0.04). CFSE carboxyfluorescein succinimidyl ester, DC dendritic cell, HA hemophilia A, IL-10 interleukin-10, n number of experiments, nT reg naïve regulatory T, sT reg sensitized regulatory T, T reg regulatory T (***) P < 0.0001, (*) P < 0.05