An improved method for the establishment of a model of Graves' disease in BALB/c mice

Abstract

The present study aimed to develop a stable Graves' disease (GD) model in BALB/c mice by immunization and electroporation (EP). A total of 90 mice were divided into experimental (n=50), control (n=20) and blank (n=20) groups. The recombinant plasmid pcDNA3.1/thyroid-stimulating hormone (TSH) receptor 268 was constructed and injected into the bilateral gastrocnemius of experimental group mice at weeks 1, 4, 7 and 10. Equal volumes of saline were injected into the control and blank groups at the same time. The experimental and control groups were subjected to EP at the same time and location to enhance immunization. The levels of total serum thyroxine (T4) and serum TSH were examined by radioimmunoassay and immunoradiometric assay, respectively. The levels of serum thyrotropin receptor N-terminal (TRAb N) and C-terminal (TRAb C) antibodies were assessed by ELISA. Whole body pertechnetate (99mTcO4-) imaging was performed. Mouse weight and thyroid morphology and pathology were analyzed. The GD BALB/c mouse model was successfully established, with a positive rate of 79.17% (38/48). T4 levels increased from baseline levels of 12.05±4.23 to 52.51±23.58 ng/ml by week 12 (P<0.0001). TSH levels decreased from baseline levels of 5.53±2.78 to 1.43±0.89 µIU/ml by week 12 (P<0.0001). TRAb N antibody levels increased from baseline levels of 0.006±0.002 to 0.278±0.106 mIU/ml by week 12 (P<0.0001). TRAb C antibody levels increased from baseline levels of 11.111±2.808 to 46.701±26.436 arbitrary units/ml by week 12 (P<0.0001). At week 21, TSH levels remained reduced compared with pre-immunization levels (P<0.0001). Although T4, and TRAb N and C levels decreased, they remained increased compared with preimmunization levels (P<0.0001, P<0.0001, P=0.001). There were no significant alterations in antibody levels between the control and blank groups. Following four immunizations, the uptake of 99mTcO4- by the thyroid was significantly increased in the experimental group. The mean weight of the experimental mice was significantly reduced compared with the control and blank groups (all P<0.0001). Furthermore, the thyroid glands of the immunized mice were enlarged and exhibited lymphocyte infiltration, fewer colloid nodules and an increased height of epithelial cells. In conclusion, by injecting recombinant plasmid pcDNA3.1/TSHR268 and EP, a GD mouse model was successfully established.

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

“…The incidence of the disease is an important sign to evaluate the successful of a model that thyroid alterations and hyperthyroidism were the key factors in the GD model [15]. In 2002, the incidence of GD in BALB/c mice was about 50-60% [8]. Some researchers improved the incidence of GD by combining Ad-TSHR289 immunization with other methods.…”

“…DCs infected with AdTSHR289 immunized in BALB/c mice with a disease induction rate of about 70% [16]. 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].…”

“…In the recent two decades, some approaches for GD models with adenovirus have been demonstrated. Combining adenovirus with electroporation or Dentritic Cells (DCs) was more effective than before [7,8]. What's more, the rate of GD model could reach 70%-100% only using recombinant adenovirus expressing the full-length human TSHR [9,10].…”

Immunotherapy with a biologically active ICAM-1 mAb and an siRNA targeting TSHR in a BALB/c mouse model of Graves’ disease

“…All 70 female BALB/c mice were injected with pcDNA 3.1/TSHR 289 into the bilateral gastrocnemius and electroporated (EP) every 3 weeks 4 times. The protocol for establishing the GD mouse model was described in our previous article [13]. This work was approved by the Ethics Committee of Tianjin Medical University General Hospital (NO.…”