Figure 6.

Combination therapy with suboptimal or optimal doses of clozapine and glatiramer acetate provided enhanced disease reduction compared to monotherapy. (a) RAW macrophages were primed with IFN-γ for 18 hours prior to stimulation with LPS (200 ng/mL) in the presence or absence of clozapine (10 µM) and increasing concentrations of glatiramer acetate for 24 h. Viability was assessed by MTT assay and IL-12p40 by ELISA. Shown are the means and SEM from two or three independent experiments. *p < 0.05 and ***p < 0.001 by 2-way ANOVA with Bonferroni’s post-test. (b–d) C57BL/6 mice were immunized and treated with clozapine (10, 30, or 60 mg/kg/day in drinking water from day -1) or vehicle as in Figure 1(a) and glatiramer acetate (100 or 500 µg/mouse) was included in the EAE immunization at d0. Mice were weighed and scored daily and cumulative disease assessed by AUC and expressed as % vehicle. Shown are the means and SEM from individual mice (n = 15–29/group) from at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by 1-way ANOVA with Tukey’s multiple comparison test.