Method Article

Induction and Clinical Scoring of Chronic-Relapsing Experimental Autoimmune Encephalomyelitis

DOI:

10.3791/224

July 4th, 2007

In This Article

Summary

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This video demonstrates the induction and clinical scoring of an animal model of multiple sclerosis: chronic-relapsing experimental autoimmune encephalomyelitis in DA rats. The disease, induced by immunizing rats with an emulsion containing whole rat spinal cord and complete Freund's adjuvant, presents clinical signs resembling the human disease.

Abstract

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Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) that commonly affects young adults. It is characterized by demyelination and glial scaring in areas disseminated in the brain and spinal cord. These lesions alter nerve conduction and induce the disabling neurological deficits that vary with the location of the demyelinated plaques in the CNS (e.g. paraparesis, paralysis, blindness, incontinence).

Experimental autoimmune encephalomyelitis (EAE) is a model for MS. EAE was first induced accidentally in humans during vaccination against rabies, using viruses grown on rabbit spinal cords. Residues of spinal injected with the inactivated virus induced the CNS disease. Following these observations, a first model of EAE was described in non-human primates immunized with a CNS homogenate by Rivers and Schwenther in 1935. EAE has since been generated in a variety of species and can follow different courses depending on the species/strain and immunizing antigen used. For example, immunizing Lewis rats with myelin basic protein in emulsion with adjuvant induces an acute model of EAE, while the same antigen induces a chronic disease in guinea pigs.

The EAE model described here is induced by immunizing DA rats against DA rat spinal cord in emulsion in complete Freund's adjuvant. Rats develop an ascending flaccid paralysis within 7-14 days post-immunization. Clinical signs follow a relapsing-remitting course over several weeks. Pathology shows large immune infiltrates in the CNS and demyelination plaques. Special considerations for taking care for animals with EAE are described at the end of the video.

Protocol

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Induction and monitoring of chronic-relapsing EAE

The emulsion is a 1:1 mixture of antigen in aqueous solution added to supplemented complete Freund's adjuvant. It is very important to add the antigen to the adjuvant and not the other way round!!!

There is a lot of waste when making and injecting an emulsion. Always prepare 1.5 or 2 times more than what you need.

Use autoclaved mortar, pestle, glass syringes, and bridges. All plastics should be sterile. The preparation can be done on a regular laboratory bench.

1. Preparatio....

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Discussion

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There are a few important considerations for this protocol.

Genetics of DA rats will differ slightly depending on which breeder they are bought from. These differences may increase or decrease the susceptibility of rats to EAE induction. If your rats are more susceptible to EAE you will want to reduce the strength of the immunization by using non-supplemented complete Freund's adjuvant or even incomplete Freund's adjuvant. You may also consider reducing the amount of emulsion injected or dilu.......

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Disclosures

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CB and KGC are co-founders and consultants for Airmid Inc.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Complete Freund’s AdjuvantReagentFisher ScientificDF 311-360-5Manufactured by Difco
Mycobacterium tuberculosis H37RaReagentFisher ScientificDF3114-33-8Manufactured by Difco
DA ratAnimalHarlan LaboratoriesFemales, 7-9 weeks old.
Glass syringes - 5 mlToolFisher Scientific14-823-10 B
Metal bridge - 18GToolFisher Scientific14-825-17L
Luer Lok plastic syringes - 3 mlToolBD Biosciences309585
Needles - 20G 1 1/2ToolFisher Scientific14-826-5C

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Tags

Experimental Autoimmune EncephalomyelitisChronic Relapsing EAEDA Rat ModelSpinal Cord HomogenateComplete Freunds AdjuvantClinical Scoring SystemAscending Flaccid ParalysisRelapsing Remitting CourseCNS DemyelinationAnimal Model MS

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