Generation of Experimental Autoimmune Encephalomyelitis in a Mouse Model

Inject the myelin-derived peptides with an adjuvant near the peripheral lymph nodes of an anesthetized mouse.

The peptide-adjuvant conjugates activate the antigen-presenting cells, or APCs, which recognize and process the peptides.

The APCs then migrate to the lymph nodes and activate the autoreactive T cells.

Periodically inject the pertussis toxin intraperitoneally, which disrupts the tight junctions between the endothelial cells and increases the blood-brain barrier permeability.

The autoreactive T cells circulate in the blood and cross the blood-brain barrier to enter the brain tissue.

These cells recognize the peptides on the myelin sheath and release the cytokines.

These cytokines recruit macrophages and B cells to the site.

Macrophages release reactive oxygen species that damage the myelin-producing oligodendrocytes, while the B cells interact with T cells, become activated, and produce autoantibodies targeting the myelin sheath.

This leads to demyelination, impairing nerve transmission.

Lift the mouse tail; a gradual drop indicates nerve damage, signaling the onset of autoimmune encephalomyelitis, or EAE.

Begin this procedure with anesthetization of C57BL/6 mice that have been housed in specific pathogen-free conditions as described in the text protocol. Fix the anesthetized mouse in one hand and inject 30 microliters of MOG peptide in complete fluorescent adjuvant emulsion subcutaneously into each of the hind leg flanks in close proximity to the inguinal lymph nodes.

Now, place the mouse on its belly and inject 20 microliters of the MOG peptide emulsion into the soft fatty tissue on both the left and right at the tail root. Also, inject a little droplet of the emulsion into the neck of the mouse. Inject 100 microliters of pertussis toxin solution intraperitoneally into the mouse. Hold the head of the mouse below the body center to avoid injection into the intestine.

Replace the maintenance diet to the breeding diet in order to provide the mice with food of higher energy content before enduring the expected clinical disease. Repeat the pertussis toxin injection 48 hours after the first treatment.

Check the health status of EAE mice every morning by taking a look inside the cages. Score EAE mice every afternoon. Take every individual mouse included in the EAE experiment out of the cage, and check whether the tail has tonus.

Move the tail upward with a finger. A healthy mouse will keep its tail up. If clinical EAE has started, the tail tonus will be lower visible by a gradual drop of the tail. Eventually, the mouse will not be able to lift its tail at all.