This article outlines a method for inducing experimental autoimmune encephalomyelitis (EAE) in mice, a model for studying multiple sclerosis. The procedure involves the injection of a neuronal peptide and a bacterial adjuvant to stimulate an autoimmune response.
Begin by subcutaneously injecting an emulsion containing a neuronal peptide and a heat-killed bacterial adjuvant into an anesthetized mouse.
In the subcutaneous tissue, the adjuvant attracts antigen-presenting cells, or APCs, which process and present the neuronal peptides on major histocompatibility complexes.
The activated APCs reach the lymph nodes and interact with naïve T cells, inducing their differentiation into autoreactive T cells.
These T cells circulate and reach the blood-brain barrier or BBB.
Next, intraperitoneally inject bacteria-derived exotoxins.
The exotoxins reach the BBB, increasing its permeability and allowing T cell infiltration into the central nervous system (CNS).
In the CNS, T cells interact with resident APCs presenting the neuronal peptide, promoting cytokine release that induces further immune cell infiltration.
The immune cells release pro-inflammatory cytokines, causing myelin damage.
Additionally, B-cells interact with T cells and secrete autoantibodies that further damage myelin, establishing experimental autoimmune encephalomyelitis (EAE).
In this procedure, dissolve PTX at 2 micrograms per milliliter in 1x PBS, and mix well. Then, inject 100 microliters of PLP/CFA in the mice subcutaneously, twice at the base of the tail during short isoflurane anesthesia. Do not inject into the back of the neck, because any immune reactions in the skin in the upper back or neck will disturb imaging of the head and spinal cord.
One to two hours after immunization, inject 100 microliters of PTX in the mice intraperitoneally.
Then, 24 hours after immunization, inject the mice with 100 microliters of PTX again. For the control mice, inject 100 microliters of CFA without PLP twice, at the base of the tail.
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