A2AR engagement during inflammation inhibits IL-6 and promotes Foxp3 expression in vivo. (A,B) Primary T cells from 5C.C7 mice were cultured with PCC plus or minus 1 μM CGS for 1 day. Total RNA was harvested and assayed for abundance of IL-6 (A) and TGF-β (B) transcripts by real-time PCR. Data are representative of 3 independent experiments. (C) Splenocytes from 5C.C7 TCR-transgenic Rag2−/− mice were cultured with antigen (PCC) plus or minus 1 μM CGS for 3 days under Th17-driving conditions (containing TGF-β, IL-6, anti-IFN-γ, and anti-IL-4). The cells were then activated with PMA and ionomycin and assayed for IL-17 by intracellular cytokine staining. Representative FACs plots are shown (left) and bar graph of composite data (right). Data are representative of 3 independent experiments. (D) Primary T cells from 5C.C7 mice were cultured with PCC plus or minus 1 μM CGS for 1 day. Total RNA was harvested and assayed for abundance of Foxp3 transcripts by real-time PCR. Data are representative of 3 independent experiments. (E) 6.5+ donor T cells were harvested 3 days after transfer into C3HA hosts treated with vehicle or CGS. Total RNA was harvested and assayed for abundance of Foxp3 by real-time PCR. Data are representative of 2 independent experiments, 3 mice per group. All PCR samples were done in triplicate and evaluated for significance (*P < .05).
