Fig. 7

Paralysis severity and neurodegeneration in the anterior visual pathway in immunization EAE mice treated after paralysis onset with LABP-66. (A) Wild-type 12-week-old female mice were immunized against MOG_{35 − 55} to induce EAE. After the development of paralysis (EAE score 1.0 or greater), mice were randomized to daily oral gavage with LABP-66 (20 mg/kg; n = 22) or vehicle (0.5% methylcellulose; n = 24). At 42 days post-immunization mice were euthanized, and retina and optic nerves were isolated. (B) Serum concentration of LABP-66 2–4 h after either LABP-66 (20 mg/kg; n = 8) or vehicle (n = 5) oral gavage in WT EAE mice. The data was normalized to the assay limit of detection (LOD). Data presented as mean ± SEM with statistical analysis performed by Mann-Whitney U test. (C) EAE paralysis clinical score overtime normalized to day post randomization in LABP-66, and vehicle treated EAE mice with statistical analysis presented representing area under the curve analysis and acute and chronic time points. (D) EAE paralysis score area under the curve (AUC) analysis for days 15–24 post randomization. (E) Change in EAE paralysis score from peak score to lowest score post peak. Data for AUC and change in EAE score are presented as median ± interquartile range (IQR). (F) Representative IHC for Brn3a⁺ RGCs in the retina with (G) quantification of Brn3a⁺ RGC density vehicle- (n = 20) and LABP-66-treated (n = 23) EAE mice as well as naïve mice (n = 5 and CFA/PTX-only mice (n = 5). (H) Representative immunofluorescence (IF) images of neurofilament light chain (NFL) and average NFL percent area positive in the optic nerve of EAE mice treated with vehicle or LABP-66. (I) Correlation between percent area positive of NFL in the optic nerve to Brn3a⁺ RGC cell body density in the retina at day 42 post immunization in vehicle-treated and LABP-66 treated EAE mice, Correlation analysis was performed by Pearson correlation test. RGC density is presented as mean ± SEM with statistical analysis performed by one-way ANOVA with Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001