Fig. 2

Prophylactic and semi-therapeutic ibrutinib treatment in Biozzi EAE ameliorate clinical symptoms, suppress relapses, and disrupt B cell maturation. Mice were treated daily through Day 35 with vehicle or ibrutinib beginning on Day 0, or ibrutinib beginning on Day 9. Mice treated with ibrutinib from Day 9 received vehicle from Day 0 through Day 8 to control for stress. Clinical EAE analyses (a–c) were performed on n = 13 mice/group, and flow cytometric analyses (d–l) were performed on n = 7–8 representative mice/group from the same experiment. The figure legend in a applies to all graphs. a EAE clinical scores over time. Arrows indicate the final day of the study (Day 36). b Maximum EAE scores for the duration of the study. c Kaplan–Meier survival curves for day of relapse onset. d–l Spleens were collected and splenocyte suspensions generated on Day 36. d Representative dot plots of immature and mature B cells. e Proportions of B cells that were immature or mature. f Numbers of immature and mature B cells. g Representative dot plots with T1, T2, and T3 B cells. h Proportions of immature B cells that were T1, T2, or T3 B cells. i Numbers of T1, T2, and T3 B cells. j Representative dot plots with marginal zone (MZ) and follicular (FO) B cells. k Proportions of mature B cells that were MZ or FO B cells. l Numbers of MZ and FO B cells. Except in a showing mean + SEM and c showing total percentage per group over time, data are shown as mean ± SEM. Significance was tested using a Kruskal–Wallis test followed by Dunn’s multiple comparisons test (end score and maximum score), Gehan–Breslow–Wilcoxon tests with pvalues adjusted for multiplicity using the Holm-Sidak approach (time to relapse onset), or one-way ANOVA followed by Dunnett’s multiple comparisons test (flow cytometric analyses). Asterisks denote results of multiple comparisons tests. *p < 0.050, **p < 0.010, ***p < 0.001