Experimental Autoimmune Encephalomyelitis in the Mouse

Experimental autoimmune encephalomyelitis (EAE) is a CD4+ T cell-mediated autoimmune disease. The disease is characterized by perivascular CD4+ T cell and mononuclear cell inflammation and subsequent primary demyelination of axonal tracks in the central nervous system, leading to progressive hind-limb paralysis. EAE provides a powerful model for the study of the pathogenesis and immune regulation of CD4+ TH1-mediated tissue damage and is generally considered to be a relevant model for the human immunemediated demyelinating disease multiple sclerosis. In the SJL (H-2s) mouse, the disease is characterized by a relapsing-remitting course of paralysis, which allows assessment of the efficacy of various immunoregulatory strategies in a progressive autoimmune disease setting. In other mouse strains, such as PL/J (H-2u) and B10.PL (H-2u), the disease is normally acute and self-limiting, and is not characterized by clinical relapses.

Actively induced EAE consists of an induction phase and an effector phase. The induction phase of the disease involves the priming of myelin epitope-specific CD4+ T cells following immunization with myelin proteins or peptides in complete Freund's adjuvant. The effector phase consists of multiple stages: (1) migration of activated myelin-specific T cells to the central nervous system (CNS), which involves extravasation of the T cells across the tight endothelial junctions comprising the blood-brain barrier; (2) elaboration of chemokines and cytokines by the myelin-specific T cells, which induce the influx of peripheral mononuclear phagocytes into the CNS parenchyma; (3) activation of peripheral monocytes/macrophages and CNS-resident microglial cells by T cell-derived cytokines; and (4) demyelination of CNS axonal tracts by the phagocytic activity of activated mononuclear cells and by the cytotoxic effects of cytokines (e.g., IFN-y, LT/TNF-P, TNF-a, and NO) released from activated CD4+ T cells and monocytes. The effector phase of the disease is modeled by the adoptive-transfer model of EAE, in which disease is induced by the peripheral introduction of a preactivated population of myelin epitope-specific CD4+ T cells to a naive mouse.

This unit details the materials and methods required for both active induction (see Basic Protocol) and adoptive transfer (see Alternate Protocol) of EAE in the SJL mouse strain using intact proteins or peptides from the two major myelin proteins—proteolipid protein (PLP) and myelin basic protein (MBP). Detailed materials and methods required for the purification of both PLP and MBP are also described (see Support Protocols 1 and 2). It should be cautioned that modifications of the specified protocols may be necessary for efficient induction of active or adoptive EAE in other mouse strains, and the reader is urged to consult the literature for details (also see Commentary for more discussion).

Diabetes Sustenance

Diabetes Sustenance

Get All The Support And Guidance You Need To Be A Success At Dealing With Diabetes The Healthy Way. This Book Is One Of The Most Valuable Resources In The World When It Comes To Learning How Nutritional Supplements Can Control Sugar Levels.

Get My Free Ebook

Post a comment