Two adjuvants, bacterial lipopolysaccharide (LPS) and supplemented complete Freund's adjuvant (CFA; see Alternate Protocol), are widely used to facilitate induction of experimental autoimmune thyroiditis (EAT) with mouse thyroglobulin (MTg). These adjuvants are particularly useful if spleen cells (LPS) and lymph node cells (CFA) are to be used for adoptive transfer (see Basic Protocol 2). LPS is preferred to CFA because there is no antigen loss associated with preparing an emulsion, and the use of LPS as an adjuvant results in more uniform disease induction (see Background Information). Moreover, two injections of MTg are generally required to induce EAT, and some institutional animal care and use committees (IACUC) do not permit two CFA doses per animal.
Of the three parameters to evaluate EAT, the most important is histologic evaluation of mononuclear cell infiltration of the thyroid gland. Second is the in vitro T cell proliferative response to MTg (see Support Protocol 2), which can be an adjunct to histology. Antibody levels are least important because both susceptible and resistant mice immunized with MTg and LPS or CFA make antibodies to MTg. The presence of antibody to MTg indicates only that an autoimmune response has occurred.
Contributed by Yi-chi M. Kong
Current Protocols in Immunology (1996) 15.7.1-15.7.16 Copyright © 1996 by John Wiley & Sons, Inc.
Was this article helpful?