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Fig. 6 | Journal of Translational Medicine

Fig. 6

From: Autoantigen-specific immunosuppression with tolerogenic peripheral blood cells prevents relapses in a mouse model of relapsing-remitting multiple sclerosis

Fig. 6

Treatment of EAE mice with MICCop increases CD4+CD25+FoxP3+ cell infiltration of peripheral lymphoid organs and the central nervous system. The infiltration with CD4+CD25+FoxP3+ regulatory T cells (Tregs) of peripheral lymphoid organs (a and b) and the central nervous system (c and d) of cell-treated and control EAE mice was determined with FACS analysis in SPCs and immunohistochemistry in spinal cord sections, respectively. a For FACS analysis SPCs of animals treated with PBS (control), MICCop and UVC-SPCCop were isolated 4 weeks after cell therapy and stained with fluorescence-labeled antibodies and the corresponding isotype controls. One representative FACS-plot is shown for each group representing the expression of CD25 and FoxP3 in CD4+ cells. b Tregs within SPCs of each treatment group were quantified and are depicted as percentage of CD4+CD25+FoxP3+ T cells within the CD4+ T-cell population (mean value ± SEM; n = 8 per group). Data were statistically compared with the unpaired Student’s t test (*p < 0.05). c Immunohistochemistry of the lumbar spinal cord for evaluation of infiltrating Tregs was performed on snap-frozen tissue of PBS- and MICCop-treated animals. After incubation of tissue sections with anti-FoxP3-antibody, Tregs were visualized with biotin-conjugated rabbit anti-rat IgG and Neutravidin-Dylight549. Cellular nuclei were counterstained with Hoechst 33,342 (blue). Digital fluorescence images were obtained at ×40 magnification and the number of infiltrated Tregs (pink) was determined. Scale bars depict 50 µm. d Statistical analysis of evaluated sections based on values from single animals (n = 8 for each group) was performed using Mann–Whitney U test (*p < 0.05). Mean values of FoxP3+ cells per mm2 ± SEM are presented

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