Induction Of Active Eae With Myelin Basic Protein

In this protocol, rats are injected with guinea pig myelin basic protein (MBP) emulsified in a modified complete Freund's adjuvant (CFA) to induce active experimental autoimmune encephalomyelitis (EAE), which becomes evident within 10 to 12 days. Commercial complete Freund's adjuvant (CFA) does not contain a sufficient quantity of mycobacteria to elicit reproducible severe EAE. Consequently, CFA is prepared by supplementing incomplete adjuvant with mycobacteria. Either Lewis (LEW) or Dark Agouti (DA) rats may be used, although the former are recommended, because DA rats also tend to develop arthritis in response to the CFA.

Materials

Lyophilized guinea pig MBP (see Support Protocol 1) Heat-killed Mycobacterium butyricum (Difco) Incomplete Freund's adjuvant (IFA, Difco)

LEW rats, preferably 8- to 10-week-old pathogen-free females (Charles River Labs)

12 x 75-mm test tube

1-ml Luer-Lok tuberculin syringe

18- and 26-G needles

Additional reagents and equipment for footpad injection (unit 1.6)

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