Twenty-four hours after injury or sham treatment, CD3⁺ T-cells were isolated from pLNs and were adoptively transferred into naive mice together with CFSE-labelled OVA-specific T-cells from DO11.10 mice. OVA was injected into the hind footpads and pLN cells were isolated and pooled per group 3 days later. (A) Overview of the experimental design. (B) Content of IFN-γ, IL-10 and IL-2 in the supernatants of pLN cells after restimulation with pOVA. Data show mean ± S.D. of triplicate cultures from pLN cells and are representative of three independent experiments (n=3 mice each). Statistically significant differences between sham-treated mice and injured mice were detected with Student's t-test. *P<0.05; **P<0.01; ***P<0.001. (C and D) The pLN cells were isolated 24 h after injury or sham treatment (no transfer of OVA-specific T-cells). (C) Dot plots show the expression of CD25 and CD69 by gated CD4⁺ T-cells. The numbers indicate the percentage within the respective quadrants. (D) FoxP3 expression in gated CD4⁺ T-cells. Bar graph shows the mean percentage of FoxP3⁺ among CD4⁺ Th-cells from pLN cells of individual mice (n=3 mice each). Data are representative of two experiments.