EAE Induction and Therapeutic Interventions

EAE was induced in eight to ten weeks old mice by subcutaneous immunization with 200 μg MOG_35–55 peptide emulsified in complete Freund’s adjuvant (CFA, Difco Laboratories) per mouse, followed by administration of 200 ng pertussis toxin (PTX, List biological laboratories, Inc.) on days 0 and 2 as described^{26 (link)}. Clinical signs of EAE were assessed according to the following score: 0, no signs of disease; 1, loss of tone in the tail; 2, hind limb paresis; 3, hind limb paralysis; 4, tetraplegia; 5, moribund. Human Interferon-beta 1a (Rebif, Merck Serono) or vehicle control was administered daily at a dose of 5.000 IU intranasally or intraperitoneally as outlined in the specific experiments. Antibiotics, Trp-indoles, and TnAse were administered daily by oral gavage starting from day 22 after EAE induction at the following doses: Ampicillin 6 mg/20 g body weight (BW), vancomycin 3 mg/20 g BW; indole, indole-3-propionic acid, indole-3-aldehyde at 400 μg/20g BW, TnAse at 200 μg/20 g BW. I3S was administered daily intraperitoneally at a dose of 200 μg/20g BW. All agents were purchased from Sigma Aldrich.