Fig. 5

In vitro treatment of T cells with decidua-derived mesenchymal stem cells (DMSC) interferes with Th17 phenotype definition. Spleen cells (SC) from C57BL/6 mice were stimulated in vitro by anti-CD3/anti-CD28 antibodies under nonpolarizing condition (a, c) or under pro-Th17 pressure in the presence of IL-6 and TGFβ (b, d, e). Three-day cultures were analyzed for IL-17 expression (a, b) or used for a second round of anti-CD3/anti-CD28 stimulation after DMSC removal during 3 subsequent days before new analysis (c, d, e). Soluble IL-17 measurements were quantified by ELISA (a–d). Each sample was assayed in quintuplicate and significance was analyzed by t-test; standard error of the means are shown. Percentages of IL-17⁺ (e) or RORγT⁺ (f) cells were determined by FACS analysis of intracellular staining with anti-IL17 and anti-RORγT antibodies, respectively (e). Results are shown from one representative out of three independent experiments. *p < 0.05, ***p < 0.001. IL Interleukin, Th T helper