Mice were immunized subcutaneously with 100 µl of emulsified complete Freund’s adjuvant (CFA, Sigma) supplemented with 10 µg/µl Mycobacterium tuberculosis and 100 µg MOG35–55 peptide (Charité Berlin) and received intraperitoneal injections of 200 ng pertussis toxin (List Labs) at the time of immunization and 48 h later. Mice were monitored every other day for signs of clinical manifestation of EAE, and clinical scores were attributed as follows: 0, no disease; 1, tail weakness; 2, hind limb weakness; 3, hind limb paralysis; 4, hind limb paralysis with forelimb weakness; 5, moribund or dead animals.
