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. 2020 Nov 19;11:575451. doi: 10.3389/fimmu.2020.575451

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Copyright © 2020 Dagkonaki, Avloniti, Evangelidou, Papazian, Kanistras, Tseveleki, Lampros, Tselios, Jensen, Möbius, Ruhwedel, Androutsou, Matsoukas, Anagnostouli, Lassmann and Probert

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Therapeutic OM-MOG reverses spinal cord neuropathology in DR2b.Ab° mice, and increases the expression of myelin and neuronal genes, and preserves axons in B6 mice during MOG-EAE. (A–D) Neuropathological analysis of spinal cord sections from therapeutic vehicle- (left panels of each photo) and OM-MOG-injected (right panels of each photo) DR2b.Ab° mice on dpi 20 for EAE. (A, D) Inflammatory cell infiltration was measured by H&E; (B, D) demyelination by LFB; (C, D) microgliosis by Iba1 immunostaining; (D) axon damage by amyloid precursor protein (APP) immunostaining. Vehicle-treated control mice show large confluent inflammatory, demyelinating lesions with neuroinflammation typical of MOG-EAE (arrowheads). OM-MOG-treated mice showed markedly reduced spinal cord pathology. Representative sections from 1 of 5 (OM-MOG) or 4 (vehicle) mice per group are shown. Scale bar 500 μM. (E) Quantitative RT-PCR analysis of mRNA levels for selected markers, H2-Ab1 (inflammation), Mbp (myelin) and Snap25 (neurons), relative to Gusb, in whole spinal cord recovered from B6 mice before immunization for MOG-EAE (non-EAE, n=4) or during MOG-EAE on the day of the first clinical symptom (onset, n=4) and after 5 injections of vehicle (n=5; for H2-Ab1, n=4) or OM-MOG (n=5; for H2-Ab1 n=4). (F–H) Representative electron micrographs of anterior funiculus of lumbar spinal cord, a major site of inflammatory lesions, from B6 EAE mice before treatment (clinical score 2, “onset”) (F), or after 5 i.d. injections of vehicle (G) or OM-MOG (H). White arrowheads show collapsed axons, black arrowheads show remyelinated axons, scale bar 5 μM. (I) Percentages of non-collapsed myelinated axons (left graph) and collapsed axons (middle graph), and quantification of the g-ratios (axon diameter/fiber diameter) of myelinated axons (right graph) in the same groups of mice shown in (F–H) (n=3 per group). Therapeutic effects of OM-MOG at the clinical level were seen in one (DR2b.Ab° mice) or more (B6) independent ËAE experiments. Data and statistical significance for each analysis are derived from one experiment each, after pairwise comparisons between samples from different mouse groups using Student’s t test (D, E), Kruskal-Wallis test (I, axon measurements) or one-way ANOVA (I, g-ratios) (*p ≤ 0.05, **p ≤ 0.01 ***p ≤ 0.001).