EAE Induction

Healthy C57BL/6 mice (15 weeks old) were immunized subcutaneously at the base of the tail with a total of 300 μg recombinant human myelin oligodendrocyte glycoprotein (rhMOG), [extracellular domain (1–125) produced by SinoBiologics in Escherichia Coli and was supplied by Novartis Institute for BioMedical Research Switzerland], emulsified in incomplete Freund's adjuvant, supplemented with 4 mg/mL of Mycobacterium tuberculosis. The mice received an intra-peritoneal injection of 150 ng of toxic protein from Bordetella pertussis in saline at the time of immunization and 48 h later. The control mice were subjected to the same procedure as the EAE-induced mice, except that rhMOG was replaced with saline (sham-injected). EAE induction was performed in a total of 39 EAE mice and 18 control mice. The mice were weighed and examined daily for clinical signs of EAE using standard scoring (0, no paralysis; 1, loss of tail tone; 2, hind limb weakness or paresis; 3, hind limb paralysis; 4, hind limb paralysis and forelimb paresis; 5, moribund or deceased).