Fig. 7

Blockade of DLL4 protects against blood-brain barrier opening and paralysis in EAE. (A) Mean clinical scores are shown from mice (10-week-old female C57BL/6, 9 per group) sensitized with EAE MOG_35–55, then from onset of disease (8 days post-induction) treated every 3 days for 11 days (red arrows) with an inVivoMab anti-mouse DLL4 antibody (500 µg/mouse/d) or an inVivoMab polyclonal Armenian hamster IgG (500 µg/mouse/d) in the control group. Neurological deficit was scored (from day 1 to day 25 post sensitization) daily for each animal according to a widely-used 5-point scale (EAE scoring: 1 limp tail; 2 limp tail and weakness of hind limb; 3 limp tail and complete paralysis of hind legs; 4 limp tail, complete hind leg and partial front leg paralysis), Statistical analysis was determined by using a 2 ways Anova test followed by the Holm-Sidak’s multiple comparisons test (*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001 ****: p ≤ 0.0001). (B) For each group of mice, EAE peak score and (C) EAE average score during time of disability was quantified other the course of the disease. (D-I) Cortical lesions were immuno-stained with (D) anti MBP (in green) and anti-IBA1 (in red) antibodies, (E) anti-CD4 (in green) and anti-CDH5 (in red) antibodies, (F) anti-IgG (in grey) and anti-CDH5 (in red) antibodies, (G) anti-GFAP (in green) and anti-IL-6 (in red) antibodies, (H) anti-CD31 (in green), anti-VIM (in red) and anti-TYMP (in grey) antibodies and (I) anti-VEGFA (in green) and anti-GFAP (in red) antibodies. (D-I) Nuclei were stained with DAPI (in blue). (J) MBP, (K) CD4, (L) IBA1, (M) IgG, (N) VIM+/CD31-, (O) GFAP, (P) IL-6, (Q) TYMP and (R) VEGFA positive areas were quantified in spinal cord lesions (3 lesions per tissue). (inVivoMab anti-mouse DLL4 antibody group n = 7, inVivoMab polyclonal Armenian hamster IgG n = 6). Statistical significance was determined by using a Mann-Whitney U test