Figure 3

Activated regulatory and effector CD4+ T cells move rapidly in healthy CNS tissue independent of antigen specificity. T cell brain slice co-culture experiments were performed for activated antigen-specific IL-10 producing regulatory and effector CD4+ TH2 cells as well as for polyclonally activated Foxp3 regulatory and effector CD4+ TH1 cells. (A) T cell subsets were characterized by intracellular staining of typically expressed molecules for each cell type. For detection of interleukins, T cells were stimulated by aCD3/aCD28 (1 μg/ml) prior to staining. (B) Movement patterns of polyclonally and antigen-specific activated regulatory and effector T cells were analysed by time lapse TPLSM at 80-130 μm depth after an initial invasion period of 30-45 min. For each series, migratory routes of two exemplary cells (white) are represented by white arrows. (C) Vector vessel angels of individual cell tracks of different activated T cell subsets (OT-2 Th2, OT-2/2d2 IL-10 producing regulatory T cells, C57BL6 Th1, C57BL6 Foxp3 Treg) were calculated using cell tracking analysis for individual cells of 2-6 independent experiments per group (± SD). ns: not significant, Kruskal Wallis Test.