mDCs-2D and mDCs-3D derived from C57BL/6 mice were harvested as described previously and injected (3 × 106 cells/mouse) into the C57BL/6 recipient mouse abdomen on days −6, −4, and 0. Then, splenocytes isolated from BALB/c mice were injected into the dorsal subcutaneous tissue of C57BL/6 recipient mice (1 × 107 cells/mouse) on days 0 and 3. On day 7, C57BL/6 recipient mice were challenged by injection of BALB/c splenocytes into the right hind footpad (1 × 107 cells/mouse). The left hind footpad of each recipient mouse was injected with the same amount of normal saline (NS) as a control. The negative control mice were injected with the same amount of NS. At 24 h after injection, the footpad thickness was measured using a micrometer. The extent of footpad swelling was measured by subtracting the baseline thickness of the left footpad from the thickness of the right footpad. The peripheral blood of experimental mice was collected to determine the levels of IL-10 and TGF-β1 using ELISA kits.
To determine the proportion of Treg cells, the splenocytes of experimental mice were isolated using Ficoll, and stained with FITC-labelled anti-CD4 (BD Pharmingen), APC-labelled anti-CD25 (BD Pharmingen), and PE-labelled anti-Foxp3 (BD Pharmingen). The proportion of Treg cells was analysed with BD FACSDiva Software v6.1.3.
To determine the proportion of Treg cells, the splenocytes of experimental mice were isolated using Ficoll, and stained with FITC-labelled anti-CD4 (BD Pharmingen), APC-labelled anti-CD25 (BD Pharmingen), and PE-labelled anti-Foxp3 (BD Pharmingen). The proportion of Treg cells was analysed with BD FACSDiva Software v6.1.3.