Induction of Experimental Autoimmune Encephalomyelitis in a Mouse Model

Abstract

Source: Schmitz, K., et al. Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice. J. Vis. Exp (2017).

In this video, a mouse model of multiple sclerosis is established using experimental autoimmune encephalomyelitis (EAE). The mouse is injected with an emulsion containing a neuronal peptide and an adjuvant, triggering an immune response that activates autoreactive T cells. This is followed by an injection of pertussis toxin (PTX) to increase blood-brain barrier permeability, allowing the autoreactive T-cells to infiltrate the brain. This infiltration initiates a signaling cascade that ultimately results in myelin damage, mimicking the demyelinating condition of multiple sclerosis.

Protocol

1. EAE Induction in SJL/J Mice

1. Mice
   1. Use 11-week-old female SJL/J mice and allow them to habituate to the experimental room for about 7 days. Use n = 10 mice per group.
   2. To assess medication effects, continuously administer the drug and placebo for the control group via drinking water or food pellets starting 3 or 5 days after immunization (n = 10 per group). During the peak of the disease, administer medication or placebo with milk or 3% sugar water-soaked cornflakes.
2. Immunization material
   1. Use an EAE induction kit consisting of antigen (peptide of proteolipid protein, PLP139-151, 1 mg/mL emulsion) in an emulsion with complete Freund's adjuvant (CFA, heat-killed Mycobacterium tuberculosis H37 Ra) and 2 vials (5 μg each) of lyophilized pertussis toxin (PTX).
   2. Dissolve the PTX (2 μg/mL) in 1x phosphate-buffered saline (PBS, i.e., add 1.5 mL of PBS to each PTX tube, mix well, remove with the same pipet tip, and add to 1 mL of PBS in a 50-mL tube); mix well.
3. Immunization
   1. Inject PLP/CFA subcutaneously in 2 portions, each 100 μL, both at the base of the tail.
   2. Inject 100 μL of PTX intraperitoneally (i.p.) twice per mouse, the first 1 - 2 h after immunization and the second at 24 h.
   3. For the control mice, inject CFA without PLP (2 portions of 100 μL) plus PTX without PLP.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
PLP139-151/CFA emulsion	Hooke Labs, St Lawrence, MA	EK-0123	EAE induction kit
Pertussis Toxin	Hooke Labs, St Lawrence, MA	EK-0123	EAE induction kit
IVIS Lumina Spectrum	Perkin Elmer, Inc., Waltham, USA		Bioluminescence and Infrared Imaging System
LivingImage 4.5 software	Perkin Elmer, Inc., Waltham, USA	CLS136334	IVIS analysis software
Isoflurane	Abbott Labs, Illinois, USA	26675-46-7	Anaesthetic