Fig. 7
Characterization of regulatory CD4+CD25+FoxP3+ T cells of MICCop-treated EAE mice. CD4+CD25+ regulatory T lymphocytes (Tregs) of MICCop-treated SJL/J mice were separated from LNCs and SPCs (= PBMCs) 2–3 weeks after cellular treatment via a microbead-based MACS protocol. For each animal Tregs from LNCs and SPCs were pooled. a PBMCs of PBS- (control, n = 12) or b MICCop-treated (n = 12) EAE animals were stimulated with phytohemagglutinin (PHA) and co-incubated with Tregs derived from single animals treated with MICCop (black bars) at a ratio of 5:1 (n = 12). Proliferation was determined by [3H]-thymidine incorporation. Positive control was PHA-induced T-cell proliferation of PBMCs only (100 %). Graphs show the mean relative proliferation rate (%) ± SEM in relation to the positive control. For statistical analysis One-way-ANOVA with Bonferroni correction was used (***p < 0.001; n = 12). c Tregs derived from SPCs and LNCs from two animals were pooled (in total n = 6), seeded (4 × 105/200 µL/well) and stimulated with plate-bound anti-CD3 monoclonal antibody for 48 h. IL-10 secretion was analyzed by ELISA. Stimulated conventional CD4+ T cells from the same animals served as control. Data are mean ± SEM (Student´s t test; **p < 0.01)