Induction and Scoring of EAE.

Active EAE was induced in 8–12 wk-old female C57BL/6J mice that had been allowed to acclimatize to the animal facility for at least 1 wk. MOG_35–55 peptide dissolved in PBS at a concentration of 2 mg/mL was mixed 1:1 with complete Freund’s adjuvant (prepared as described earlier) to make an emulsion. On day 0, mice were immunized by injecting 50 μL of the emulsion s.c. into each of two sites on the lateral abdomen. In addition, on day 0 and again on day 2, mice were injected intraperitoneally with 250 ng pertussis toxin. Mice were weighed and scored beginning on day 7 postimmunization. Scoring was performed in a blinded manner according to the following scale: 0, no clinical deficit; 0.5, partial loss of tail tone; 1.0, complete tail paralysis or both partial loss of tail tone plus awkward gait; 1.5, complete tail paralysis and awkward gait; 2.0, tail paralysis with hind limb weakness evidenced by foot dropping between bars of cage lid while walking; 2.5, hind limb paralysis with little to no weight-bearing on hind limbs (dragging), but with some movement possible in legs; 3.0, complete hind limb paralysis with no movement in lower limbs; 3.5, hind limb paralysis with some weakness in forelimbs; 4.0, complete tetraplegia but with some movement of head; 4.5, moribund; and 5.0, dead.