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


METHODS
A 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.


RESULTS
More 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.


CONCLUSIONS
We 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.
Department of Endocrinology, Jingling Hospital, Medical School of Nanjing University, Nanjing, P.R. China.|Center for Translational Medicine and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China.|Department of Endocrinology, Mingci Cardiovascular Hospital, Wuxi, P.R. China.
Nan
N
Xia
Department of Endocrinology, Jingling Hospital, Medical School of Nanjing University, Nanjing, P.R. China.
Xiaozhen
X
Ye
Department of Endocrinology, Jingling Hospital, Medical School of Nanjing University, Nanjing, P.R. China.
Xiaohao
X
Hu
Center for Translational Medicine and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China.
Shiyu
S
Song
Center for Translational Medicine and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China.
Hui
H
Xu
Center for Translational Medicine and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China.
Mengyuan
M
Niu
Center for Translational Medicine and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China.
Hongwei
H
Wang
Department of Endocrinology, Jingling Hospital, Medical School of Nanjing University, Nanjing, P.R. China.
Jian
J
Wang
D016475:3T3 Cells; D000818:Animals; D001323:Autoantibodies / Q000097:blood; D004195:Disease Models, Animal; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D005123:Eye / Q000000981:diagnostic imaging / Q000473:pathology; D005260:Female; D005822:Genetic Vectors; D006111:Graves Disease / Q000097:blood / Q000000981:diagnostic imaging / Q000473:pathology; D049970:Graves Ophthalmopathy / Q000097:blood / Q000000981:diagnostic imaging / Q000473:pathology; D006801:Humans; D007114:Immunization; D007273:Injections, Intramuscular; D008279:Magnetic Resonance Imaging; D051379:Mice; D008807:Mice, Inbred BALB C; D011989:Receptors, Thyrotropin / Q000235:genetics* / Q000276:immunology; D013961:Thyroid Gland / Q000473:pathology; D013974:Thyroxine / Q000097:blood
D001323:Autoantibodies; D011989:Receptors, Thyrotropin; D013974:Thyroxine