Induction and Scoring of EAE in Mice

HLA-DR3.DQ8 transgenic mice (8–12 weeks old) were immunized subcutaneously in both flanks with 25 μg of PLP₉₁₋₁₁₀ that was emulsified in Complete Freund's Adjuvant–CFA containing Mycobacterium tuberculosis H37Ra (400 μg/mouse; Becton, Dickinson and Company, Sparks, MD, USA). Pertussis toxin (PTX) (Sigma Chemicals, St. Louis, MO; 100 ng) was administered i.v. at days 0 and 2 post immunization. C57BL/6 mice were immunized subcutaneously in both flanks with MOG₃₅₋₅₅ CFA/PTX as described earlier (21 (link)). Mice were observed daily for clinical symptoms and the following scoring system was used as described previously (14 (link)): 0 for normal; 1 for loss of tail tone; 2 for hind limb weakness; 3 for hind limb paralysis; 4 for hind limb paralysis and forelimb paralysis or weakness; and 5 for morbidity/death.