Experimental Autoimmune Encephalomyelitis (EAE)

Michael K. Racke1

1 University of Texas, Southwestern Medical Center at Dallas, Dallas, Texas
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 9.7
DOI:  10.1002/0471142301.ns0907s14
Online Posting Date:  May, 2001
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The procedures described in this unit utilize murine models exclusively since murine EAE often results in a relapsing/remitting disease, similar to the early phase of most MS patients. EAE in the Lewis rat is a monophasic illness in which animals experience a single episode of paralysis from which most recover completely. This unit presents two methods for inducing EAE in mice: active induction and adoptive transfer

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Table of Contents

  • Strategic Planning
  • Basic Protocol 1: Active Incuction of EAE in Mice
  • Alternate Protocol 1: Adoptive Transfer of EAE in Mice
  • Reagents and Solutions
  • Commentary
  • Figures
  • Tables
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Basic Protocol 1: Active Incuction of EAE in Mice

  • Myelin antigen (MBP, PLP, MOG or other peptide; see Table 9.7.2). Peptides can be custom synthesized or obtained from a commercial source such as CS Bio
  • PBS ( appendix 2A)
  • Complete Freund's adjuvant (CFA; Difco)
  • Female mice from EAE susceptible strain (see Table 9.7.1), group housed 8 to 12 weeks of age
  • Methoxyflurane (anesthesia)
  • Pertussis toxin (PT; List Biological Laboratories)
  • Omni Mixer (Omni International, Popper & Sons; can also use 2‐ or 5‐ml Micro‐mate interchangeable glass syringes connected by a 7/8‐in. stainless steel, 18‐G micro‐emulsifying needle)
  • 1‐ml tuberculin syringes
  • 25‐G needles
  • Bell jar
  • Electric hair clippers
  • Animal balance (for weighing mice), accurate to 0.1 g
NOTE: Female mice are generally used because they can be group housed without difficulty. Male mice will often fight and need to be housed separately. In SJL mice, females are more susceptible to the development of EAE, but there are mouse strains (e.g., B10.PL) where males are more susceptible.

Alternate Protocol 1: Adoptive Transfer of EAE in Mice

  • Complete Hank's balanced salt solution (HBSS; see recipe)
  • Complete EAE medium (see recipe)
  • 75% ethanol
  • 50‐ml conical tubes
  • Styrofoam dissecting board
  • Surgical scissors
  • Jeweler's curved forceps
  • 4.5 × 4.5–cm stainless steel wire mesh screen
  • 60 × 15–mm petri dish, sterile
  • 3‐ml plastic syringe
  • 24‐well tissue culture plates
  • Humidified 37°C, 5% CO 2 incubator
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Literature Cited

Literature Cited
   Critchfield, J.M., Racke, M.K., Zúñiga‐Pflücker, J.C., Cannella, B., Raine, C.S., Goverman, J., and Lenardo, M.J. 1994. T cell deletion in high antigen dose therapy of autoimmune encephalomyelitis. Science 263:1139‐1143.
   Goverman, J., Woods, A., Larson, L., Weiner, L.P., Hood, L., and Zaller, D.M. 1993. Transgenic mice that express a myelin basic protein‐specific T cell receptor develop spontaneous autoimmunity. Cell 72:551‐560.
   Linington, C., Bradl, M., Lassman, H., Brunner, C., and Vass, K. 1988. Augmentation of demyelination in rat acute allergic encephalomyelitis by circulating mouse monoclonal antibodies directed against a myelin/oligodendrocyte glycoprotein. Am. J. Pathol. 130:443‐454.
   Litzenburger, T., Fassler, R., Bauer, T., Lassmann, H., Linington, C., Wekerle, H., and Iglesias, A. 1998. B lymphocytes producing demyelinating autoantibodies: Development and function in gene‐targeted transgenic mice. J. Exp. Med. 188:169‐180.
   Martin, R., McFarland, H.F., and McFarlin, D.E. 1992. Immunological aspects of demyelinating diseases. Annu. Rev. Immunol. 10:153‐187.
   Racke, M.K., Dhib‐Jalbut, S., Cannella, B., Albert, P.S., Raine, C.S., and McFarlin, D.E. 1992. Prevention and treatment of chronic relapsing experimental allergic encephalomyelitis by transforming growth factor‐beta1. J. Immunol. 154:2959‐2968.
   Ratts, R.B., Arredondo, L.R., Bittner, P., Perrin, P.J., Lovett‐Racke, A.E., and Racke, M.K. 1999. The role of CTLA‐4 in tolerance induction and T cell differentiation in experimental autoimmune encephalomyelitis: i.p. antigen administration. Int. Immunol. 11:1881‐1888.
   Zamvil, S.S. and Steinman, L. 1990. The lymphocyte in experimental allergic encephalomyelitis. Annu. Rev. Immunol. 8:579‐621.
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