Figure 6.

Conditioning ADSC with pro-inflammatory RASF enhances their ability to induce Tregs but not inhibit Th1. T cells from healthy donors were cultured in the presence or absence of ADSC at 1:40 and 1:5 ratios and activated with beads coated with anti-CD3/CD28 for 72 h. Cells were then harvested and stained with anti-CD25 and anti-Foxp3 antibodies for flow cytometry detection of Tregs (representative of 7 independent experiments) (A) or anti-IFNγ antibodies for flow cytometry detection of Th1 (representative of 6 independent experiments) (B). (C,D) ADSC were plated in 96-well plates and stimulated for 24 h with SF control, RA1, or RA12 with or without anti-IL6R and anti-TNF. The following day, ADSC were washed and T cells were added to ADSC at a ratio of 1:5 and activated with beads coated with anti-CD3/CD28 for 72 h. Cells were harvested and Tregs (C) or Th1 (D) percentages were detected by flow cytometry. Results are represented as mean ± SEM of 4–7 independent experiments. ^*p < 0.05; ^**p < 0.01.