Fig. 3

SuperMApo treatment induce pro-tolerogenic macrophages. a IL17⁺CD4⁺ (Th17), IFN-γ⁺CD4⁺ (Th1) and Foxp3⁺CD25⁺CD4⁺ (Treg) cell polarization by macrophages sorted from the spleen. Left panels show representative staining. Data from a representative experiment out of 3 showing similar results shown as mean ± SEM (5 mice per group). b Clinical score of mice treated with macrophages sorted from vehicle- (EAE + EAE_Veh-macrophages) or SuperMApo-treated mice (EAE + EAE_SMA-macrophages) injected at day 11 (arrow) post-induction (n = 4 mice). Data from a representative experiment out of 3 showing similar results, shown as mean ± SEM at each time point. c Percentages of T cell subsets in the spinal cord (left) and the inguinal lymph node (iLN) (right) seven days post-treatment of mice from b (n = 4 mice). Data pooled from 2 independent experiments shown as mean ± SEM. d Clinical score of mice treated with clodronate-loaded or PBS-loaded liposomes and then treated with vehicle or SuperMApo at day 7 and 9 (n = 3–4 mice per group). Data from a representative experiment out of 2 showing similar results, shown as mean ± SEM. The red arrow marks the injection of liposomes while the black arrows mark the injection of SuperMApo. e T cell proliferation index after MOG_35-55 peptide stimulation, from mice from d. f Th1 cell numbers of mice from d. Expression of co-stimulatory molecules g and intracellular pro-inflammatory cytokines h after LPS stimulation on macrophages during 24 h. Data from a representative experiment out of 3 showing similar results, shown as mean ± SEM. i Frequencies of Th1, Th17 and Treg cells after co-culture of naïve CD4⁺ T cells with macrophages pretreated with SuperMApo or vehicle during 24 h. Data from a representative experiment out of 2 showing similar results, shown as mean ± SEM. j Clinical score of EAE mice receiving at day 8 post-EAE induction (arrow) macrophages pretreated by SuperMApo (EAE + Macro_Med) or medium (EAE + Macro_SuperMApo) for 24 h. Data from a representative experiment out of 2 showing similar results, shown as mean ± SEM. k Numbers of CD4⁺ T cell subsets in the spinal cord (left) or iLN (right) from mice from j, 7 days after macrophage injection. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, unpaired two-tailed Student’s t test (a, c, e–i, k) or two-way ANOVA with Bonferroni posttest (b, d, j)