Fig. 6: The inflamed CNS of CD83^ΔMG exhibit disturbed T cell balance and a pro-inflammatory, disease-fostering milieu.

a, b Flow cytometric analyses of regulatory T cells (Tregs) in the inflamed CNS. Representative dot-plots (a) and quantitative assessment of the percentage of CD25+FoxP3+ among CD4+ T cells (b, n = 10 for CD83wt^Cre and n = 11 for CD83^ΔMG; pooled from two independent experiments) are shown. c, d Flow cytometric analyses of IFN-γ producing T cells in the inflamed CNS. Single cells from mice were isolated on day 18 p.i., re-stimulated with PMA/ionomycin for 5 h in the presence of Golgi transport inhibitors and intracellularly stained for IFN-γ. Representative dot-plots (c) and quantitative assessment of the percentage of IFN-γ producing CD4+ T cells (d, n = 10 for CD83wt^Cre and n = 11 for CD83^ΔMG; pooled from two independent experiments) are shown. e Expression of Th1-related transcripts in the spinal cords of EAE animals on day 18 p.i. RNA was extracted from lumbar parts of spinal cords, reversely transcribed and qPCR was performed for Ifng, Tbx21 (T-bet) and Il12a (n = 10 for CD83wt^Cre and n = 11 for CD83^ΔMG, pooled from two independent experiments). f TNF expression levels in spinal cords and sorted microglia or MDCs derived from EAE mice on day 18 p.i. (n = 10 for CD83wt^Cre and n = 11 for CD83^ΔMG; pooled from two independent experiments). g Mmp9 expression levels in spinal cords of EAE mice on day 18 p.i. (n = 10 for CD83wt^Cre and n = 11 for CD83^ΔMG; pooled from two independent experiments). Gene expression was normalized to Rpl4 and relative expression was calculated as ratio between CD83^ΔMG and CD83^wtCre. In all graphs, data are represented as mean ± SEM and Mann–Whitney U-test was used to analyze the data.