(A) We treated 2 × 10⁷ W3, W3/MOG-S, and W3/MOG-L cells with Fas ligand to induce apoptosis. Then the treated cells were intravenously injected into mice 8 days prior to immunization. Mice were then immunized with MOG_35–55 in CFA on day 0. (B) Dose-dependent effects of apoptotic cell injection on EAE. The indicated number of apoptotic W3/MOG-L cells was injected into mice on 4 days prior to immunization with MOG_35–55 in CFA on day 0. (C) Pretreatment with apoptotic cells is required for suppression of EAE. Mice were treated with 2 × 10⁷ apoptotic W3/MOG-L cells 8 or 3 days prior to immunization or 8 days afterwards. Immunization was on day 0. (D) Subcutaneous injection of apoptotic cells does not suppress EAE. We subcutaneously injected 2 × 10⁷ apoptotic W3/MOG-L cells into the tail base 6 days before mice were immunized with MOG_35–55 in CFA. (E) Intravenous injection of MOG_35–55 peptide does not induce tolerance to EAE. We intravenously injected 1 μg of MOG_35–55 peptide, which corresponds to more than 200 times the amount of MOG fragment expressed in 2 × 10⁷ apoptotic W3/MOG-L cells in molar ratio, into mice. Four days later, the mice were immunized with MOG_35–55 in CFA. (F) Secondary necrotic cells did not suppress EAE. W3/MOG-L cells were treated with Fas ligand either for 4 hours or 24 hours and were injected intravenously into mice 4 days prior to immunization with MOG_35–55 in CFA on day 0. (A–F) The disease severity of each mouse was scored, and the mean clinical scores at the indicated times were plotted. These results are representative of 3 (A–D) or 2 (E and F) independent experiments.