FIGURE 6.

ADMA treatment induces EAE disease in MOG‐immunized mice in the absence of PTX: Female C57BL/6 mice were immunized with complete Freund's adjuvant (CFA) and myelin oligodendrocyte glycoprotein (MOG) with or without pertussis toxin (PTX) or ADMA treatment. PTX was treated on days 0 and 1 of post‐immunization. ADMA (50mg/kg) was treated daily starting day 0 and ending day 10 post‐immunization. Following immunization, daily clinical scores (A‐i) and overall disease severity (AUC: area under the curve) (A‐ii) were analysed. The line and bar graphs on panel A represent mean ± standard error mean (SEM) from n = 8 animals. On day 28 post‐immunization, CNS extravasation of Evans blue (B), mononuclear cell infiltration (H&E: hematoxylin and eosin staining) and myelin status (LFB: Luxol fast blue staining) in the lumbar area of spinal cord sections (C‐i), and the number of mononuclear cells in the spinal cord (C‐ii) were analysed. Under the same experimental conditions, % of CD4⁺ T cells in CNS‐infiltrated mononuclear cells (D‐i), % of IFN‐γ⁺ T_H1 cells (D‐ii), and IL‐17⁺ T_H17 cells (D‐iii) in CNS infiltrated CD4⁺ T cells, and % of IL‐23R⁺ cells in CNS infiltrated CD4⁺ IL‐17⁺ T_H17 cells (D‐iv) were analysed by fluorescence flow cytometry. Please see Figure S4 for the representative flow cytometry dot plots. The bar graphs on panels B, C and D represent mean ± standard error mean (SEM) from n = 6 animals. N.S. (not significant: p ≥ 0·05); *p ≤ 0·05, **p ≤ 0·01, ***p ≤ 0·001 vs. control; +p ≤ 0·05, ++p ≤ 0·01, +++p ≤ 0·001 vs. as indicated