B cell composition and humoral response in EAE splenocytes isolated on day 25 following in vivo treatment of EAE mice with groups 3–9, compared to healthy and PBS negative controls: A) ELISPOT for PLP IgG antibody secreting cells (ASC) per 106 splenocytes, showing representative wells. Data represent mean ± SD (n=5). B/C) Splenocyte composition following in vivo treatment and 72h ex vivo antigen rechallenge +/− PLP (25 μM), determined by flow cytometry: B) % B cells (CD19+) overall, and C) % ABCs (CD19+B220+CD11c+) of gated B cells. Data represent mean ± SD (n=3). Statistical significance compared to PBS negative control was determined by paired t-test (A) or two-way ANOVA followed by Dunnett’s post hoc test (B,C) (*p<0.05, **p<0.01).