Simultaneous induction of Graves’ hyperthyroidism and Graves’ ophthalmopathy by TSHR genetic immunization in BALB/c mice

Abstract

BackgroundGraves’ disease is the most common form of autoimmune thyroid disorder, characterized by hyperthyroidism due to circulating autoantibodies. To address the pathological features and establish a therapeutic approach of this disease, an animal model carrying the phenotype of Graves’ disease (GD) in concert with Graves’ Ophthalmopathy (GO) will be very important. However, there are no ideal animal models that are currently available. The aim of the present study is to establish an animal model of GD and GO disease, and its pathological features were further characterized.MethodsA recombinant plasmid pcDNA3.1- T289 was constructed by inserting the TSHR A-subunit gene into the expression vector pcDNA3.1, and genetic immunization was successfully performed by intramuscular injection of the plasmid pcDNA3.1-T289 on female 8-week-old BALB/c mice. Each injection was immediately followed by in vivo electroporation using ECM830 square wave electroporator. Morphological changes of the eyes were examined using 7.0T MRI scanner. Levels of serum T4 and TSHR antibodies (TRAb) were assessed by ELISA. The pathological changes of the thyroid and orbital tissues were examined by histological staining such as H&E staining and Alcian blue staining.ResultsMore than 90% of the immunized mice spontaneously developed goiter, and about 80% of the immunized mice manifested increased serum T4 and TRAb levels, combined with hypertrophy and hyperplasia of thyroid follicles. A significantly increased synthesis of hyaluronic acid was detected in in the immunized mice compared with the control groups.ConclusionWe have successfully established an animal model manifesting Graves’ hyperthyroidism and ophthalmopathy, which provides a useful tool for future study of the pathological features and the development of novel therapies of the diseases.

An improved mouse model of Graves disease by once immunization with Ad-TSHR289

“…The method of TSHR genetic immunization and vivo electroporation was also widely used [7,8,17]. A study recently reported that over 80% of immunization mice have developed to GD and the incidence of goiter was more than 90% with immunization and electroporation [18]. However, this approach may be at risk of causing death in mice after the last immunization [7,15].…”

New advances in understanding thyroid-associated ophthalmopathy and the potential role for insulin-like growth factor-I receptor

“…They rationalize the absence of inflammatory infiltrates in their latest study as consistent with a “hit and run” immune-mediated inflammatory event 30 . Another research group, also employing intramuscular DNA immunization followed by electroporation, found that a large fraction of their animals developed elevated thyroxine levels, anti-TSHR antibodies, and goiters 31 . Extraocular muscle and adipose tissue volumes were increased.…”

Vitamin D and Selenium in a Thyroid Eye Disease Population in Texas

“…Key pathological features of TED include extraocular muscle or orbital fat expansion and soft tissue inflammation due to increased adipose tissue, oedema, and glycosaminoglycan (GAG) production in orbital tissue. 1 Left untreated, TED can cause photophobia, diplopia, pain, exposure keratopathy and proptosis. Compressive optic neuropathy threatening vision, however, is the feared complication.…”

“…Compressive optic neuropathy threatening vision, however, is the feared complication. 1 Current therapeutic options include discontinuation of smoking, correction of any thyroid abnormalities, administration of non-steroidal or steroidal antiinflammatory drugs, orbital low dose radiation therapy or surgery (e.g., orbital decompression).…”