Figure 3.

Absence of ICAM-5 worsens disease progression which can be reversed by application of sICAM-5. (A) Immunohistochemical staining of ICAM-5-AF647 on cortical neurons (scale bar = 15 μm, 3D overlay scale bar = 10 μm). Co-staining was performed with NeuN-AF488 and DAPI (blue). (B) mRNA analysis of ICAM-5 and MMP-9 were performed with cortical neurons after stimulation with LPS, IFNγ, and splenocyte supernatant (n = 6 for each condition). Unstimulated cortical neurons served as a control. (C) Immunohistochemical staining of MAP2- AF647, Tuj1-FITC, ICAM-5-AF568, and DAPI on cortical neurons (scale bar = 20 μm. (D) EAE was induced actively in ICAM-5-KO mice and WT littermates via the injection of MOG₃₅₋₅₅ (two independent EAEs, untreated; WT n = 12, KO n = 12). The dotted box on the plot provides a closer look at the chronic phase of the EAE but does not agree with the x axis of the upper plot. The lower plot shows only the mean disease score of the period from day 40 after immunization. (E) EAE lesions were stained from wt (n = 9) and ICAM5 KO mice (n = 6) for inflammation (HE staining), demyelination (LFB-PAS), and axonal loss (APP) and were quantified accordingly. (F) EAE was induced actively in B6 mice via the injection of MOG₃₅₋₅₅ and mice were treated with Methylprednisolone for 5 days as soon as the mice reached a clinical score of 2. Additionally, once animals reached a clinical score of 2, 0.2 μg ICAM-5 D1-2 Fc or human IgG was applied via intrathecal (i.th) injection by lumbar puncture seven times on every second day (WT n = 7, KO n = 7). Data shown are mean ± SEM. (C, D) Mann-Whitney U-test; *p < 0.05.