Figure 7. DCs from Lgals1^−/− MOG i.v. mice do not suppress EAE.

(A) WT (n=6) and Lgals1^−/− (n=6) mice were sacrificed 3 weeks after MOG_35-55 i.v. tolerization. CD11c⁺ DCs were isolated from splenocytes and i.v. transferred into WT EAE mice (1×10⁶ mouse; n=3–5 per group) at disease peak. Mice that received PBS-i.v. served as controls. (B) Two weeks after transfer, MNCs from the CNS of DC-transferred mice and EAE controls were isolated. MNCs were cultured overnight with MOG_35-55, activated with PMA/iono/GP, stained and analyzed by flow cytometry. Total numbers of infiltrating cells were gated from CD4⁺ population. CD4⁺IFNγ⁺, CD4⁺IL-17A⁺, CD4⁺GM-CSF⁺, CD4⁺IL-10⁺FoxP3⁺, and CD4⁺IL-10⁺FoxP3⁻ percentages in CD4⁺ T cells were determined by flow cytometry. (C) Splenocytes from MOG-i.v. and PBS-i.v. EAE WT and Lgals1^−/− mice on day 25 p.i. were stimulated for three days with MOG_35-55. Cell culture supernatants were assayed for IFN-γ, IL-17A and GM-CSF by ELISA. (D) Proliferation was measured after three days of culture with MOG_35-55. Tritiated adenine was added after 54 hours of culture; its incorporation was measured after 18 hours. Data shown are one representative of three experiments. Experiments show mean ± SD. *, p<0.05. For EAE, the area under the curve was calculated for each mouse and values for experimental groups were compared for statistical significance using a Mann-Whitney test for non-parametric data. Parametric data were compared for statistical significance using a two-tailed, unpaired Student’s t test.