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Experimental Autoimmune Encephalomyelitis (EAE)
Publication Name:
Current Protocols in Neuroscience
Unit Number:
Unit 9.7
DOI:
10.1002/0471142301.ns0907s14
Online Posting Date:
May, 2001
Abstract
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
Materials
Basic Protocol: Active Incuction of EAE in Mice Materials
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: Adoptive Transfer of EAE in Mice Additional Materials (also see
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Figures
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Figure 9.7.1 Anesthetized mice should be shaved in the areas marked by cross-hatching. These areas drain to the axillary and inguinal lymph nodes. -
Figure 9.7.2 Clinical scoring of EAE. (A) Mice that are removed from a cage will normally respond by having their tail stand straight up. When picking up a mouse by the tail, one can feel that the tail has tone. Such a mouse is normal and represents a clinical score of 0. (B) A normal mouse when placed on top of the cage will not misstep between the bars of the cage top. When a mouse has reached a score of 1, the tail no longer stands up on end. When the mouse is picked up by the tail, there is a distinct lack of tone in the tail. However, the mouse walks normally, and when put on the underside of the cage top, can climb to the top without difficulty. (C) When a mouse has reached a clinical score of 2, the mouse has a limp tail and shows signs of hindlimb dysfunction. This is most easily detected by placing the mouse on the underside of the cage, where the mouse is able to hang on, but because it does not have the same dexterity of hindlimb movement, it has difficulty climbing to the top side of the cage. When placed on the top side of the cage, the mouse may misstep and the hindlimb foot may protrude between the bars of the cage. (D) When the mouse reaches a clinical score of 3, the mouse can no longer hold on to the cage with its hindlimbs when placed on the underside of the cage. However, when ambulating, the mouse still can move the hindlimbs. (E) When the mouse reaches a clinical score of 4, the hindlimbs drag behind and are not used by the mouse for movement. A moribund mouse is still alive, but really makes little spontaneous movement and receives a clinical score of 5. These animals are routinely euthanized. -
Figure 9.7.3 Adoptive transfer of EAE, lymph node harvest. After mice have been sacrificed, the mouse is pinned on its back and opened with a midline incision. The skin is then peeled from the thorax and also pinned down. The inguinal lymph nodes are present at the junction of two draining veins. The nodes are a dusky color and should not be confused with the CFA/antigen depot, which is white. The axillary nodes are found where the arms join the trunk. One node is very superficial, and may require very little dissection in order to isolate it. The other axillary node is much deeper and requires careful dissection. This node is usually found just medial to the muscles of the upper forelimb. -
Figure 9.7.4 Activated lymph node cells from MBP/CFA immunized (Pl × SJL) F1 mice were transferred into naïve recipients at day 0. (A) Mice were monitored daily and a mean clinical score was assigned for each group of five mice. (B) Mean weights of the mice from the experiment shown in A.
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Literature Cited
| Literature Cited | |
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