Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Experimental autoimmune encephalomyelitis or EAE Model

1. Induction of the EAE model
   1. Use 10- to 13-week-old female 129S4/SvJae×C57BL/6 mice for the induction of EAE.
   2. Give the mice a subcutaneous injection, into the upper and lower back, of the encephalitogenic MOG_{35 – 55} (myelin oligodendrocyte glycoprotein) peptide (200 µg), emulsified in 200 µl complete Freund`s adjuvant (CFA) containing 400 µg Mycobacterium tuberculosis.
      NOTE: As a negative, non-disease-inducing sham control, incomplete Freund's adjuvant containing Mycobacterium tuberculosis, in the same volume and by the same route, can be used.
   3. Thereafter, and again 24 hr later, give the mice an intraperitoneal injection of pertussis toxin (in total 0.2 µg, diluted in 200 µl phosphate-buffered saline, PBS). Use untreated mice as the control group, to be able to compare diseased with healthy animals.       
      NOTE: It is also possible to induce EAE with 200 – 300 µg MOG_{35 – 55} peptide, 300 – 500 µg of Mycobacterium tuberculosis in the CFA, and 0.2 – 0.3 µg pertussis toxin in a volume of 0.1 – 0.2 ml per injection.
   4. Beginning one week after the injection, examine mice daily for clinical symptoms (see step 1.2.1)         
      NOTE: The day of disease onset varies in different experiments, but under the conditions in our laboratory, this is around day 11 and thus, here day 14 is defined as 3 days after disease onset. All mice in the present study developed clinical symptoms.

2. Scoring of the mice

1. Classify clinical symptoms by clinical scores as follows: 0) no signs, 0.5) distal paralysis of the tail, 1) complete paralysis of the tail, 1.5) limp tail and mild weakness of hind legs, 2) limp tail and severe weakness of hind legs, 2.5) limp tail and paralysis of one hind leg, 3) limp tail and paralysis of both hind legs, 3.5) paralysis of both hind limbs and weakness of one fore limb (mice achieving this score were euthanized, in keeping with local ethical guidelines).