Induction of Experimental Autoimmune Hypophysitis in SJL Mice

Summary

This video shows how to induce autoimmune hypophysitis in SJL mice and how to assess its severity by histopathology.

Abstract

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice¹. Mouse models allow us to study how diseases unfold, often providing a good replica of the same processes occurring in humans. For some autoimmune diseases, like type 1A diabetes, there are models (the NOD mouse) that spontaneously develop a disease similar to the human counterpart. For many other autoimmune diseases, however, the model needs to be induced experimentally. A common approach in this regard is to inject the mouse with a dominant antigen derived from the organ being studied. For example, investigators interested in autoimmune thyroiditis inject mice with thyroglobulin², and those interested in myasthenia gravis inject them with the acetylcholine receptor³. If the autoantigen for a particular autoimmune disease is not known, investigators inject a crude protein extract from the organ targeted by the autoimmune reaction. For autoimmune hypophysitis, the pathogenic autoantigen(s) remain to be identified⁴, and thus a crude pituitary protein preparation is used. In this video article we demonstrate how to induce experimental autoimmune hypophysitis in SJL mice.

Protocol

Experimental Protocol

Once the pituitary immunogen is prepared (see companion article), it is injected subcutaneously into SJL mice (The Jackson Laboratories, stock 000686). A second identical injection is repeated 7 days later. Mice are then used to collect in vivo and post-mortem outcomes.

Step 1. Subcutaneous injection of pituitary immunogen into SJL mice

Mice are first anesthetized by injecting 0.5 ml of 20 mg/ml 2,2,2-Tribromoethanol (Avertin, TCI America, T-1420) intraperitoneally. Mice are then injected subcutaneously with 100 μl of the pituitary emulsion that, as ....

Discussion

Four techniques have been reported to induce hypophysitis in experimental animals. The technique described in this article, that is the injection of pituitary proteins mixed with Freund s adjuvant, is the oldest one, dating back to 1967⁶, and has been improved more recently¹. Other techniques are the injection of soluble rubella virus glycoproteins in 1992⁷, the stereotactic injection of an adenovirus directly into the pituitary gland in 2002⁸, and the transplantation of pitu.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
SJL mice (The Jackson Laboratories)		stock 000686
2,2,2-Tribrom–thanol (Avertin)	TCI America	T-1420
Beckstead’s fixative