Figure 4.

OM-MOG prevents and reverses immune cell infiltration and demyelinating lesions in the spinal cord during EAE. (A) Whole mount spinal cord sections with surrounding meninges and vertebrae prepared from EAE mice after short-term prophylactic administration of vehicle or OM-MOG (dpi 7, 11 and 14) at peak of disease in the EAE group (dpi 18), and immunostained with anti-MBP for myelin (red) and anti-CD45 for immune infiltrates (green) (low power photomicrographs), or anti-laminin for blood brain barrier (LAM; red), anti-CD45 (green) and DAPI (blue) (high power photomicrographs). Representative images from 1 of 3 mice/group. Scale bars 500 μM (upper low-power images) and 100 μM (lower high-power images). (B) Whole mount spinal cord sections prepared from mice with ongoing EAE (dpi 22) after therapeutic treatment with 4 injections of vehicle or OM-MOG starting at clinical score 2, and immunostained with anti-MBP for myelin (red) and anti-CD45 for immune infiltrates (green) (low power magnification), or anti-laminin for blood brain barrier (LAM; red), anti-CD45 (green) and DAPI (blue) (high power magnification). Representative images from 1 of 3 mice/group. (C) Total numbers of splenocytes and CNS-infiltrating mononuclear cells recovered from spinal cords of EAE mice at disease peak (dpi 16) after short-term prophylactic administration, as in A (OM-MOG P), or 24 h after 1 therapeutic injection of vehicle or OM-MOG (OM-MOG T, 1 inj) (n=8 for EAE and n=4 for OM-MOG). (D) Proportions of Annexin V⁺ CNS- infiltrating mononuclear cells recovered from spinal cords of EAE mice at disease peak (dpi 14) 24 hours after 1 therapeutic injection of vehicle (EAE) or OM-MOG (OM-MOG T, 1 inj) (n=4-5/group). Data and statistical analysis are derived from one experiment in each case (A–C) or one representative of two experiments (D). Statistical significance is shown after comparison to the vehicle (EAE) group using one-way ANOVA followed by Tukey’s post hoc test (C), or Student’s t test (D) (*p ≤ 0.05, **p ≤ 0.01).