scholarly journals A Method for Identification of the Peptides That Bind to a Clone of Thyroid-Stimulating Antibodies in the Serum of Graves’ Disease Patients

2003 ◽  
Vol 88 (4) ◽  
pp. 1570-1576 ◽  
Author(s):  
Chan Hyun Na ◽  
Mi Hwa Lee ◽  
Bo Youn Cho ◽  
Chi-Bom Chae
Keyword(s):  
Graves Disease ◽  
Thyroid Stimulating Antibodies

A sensitive and practical assay for thyroid-stimulating antibodies using FRTL-5 thyroid cells

1987 ◽  
Vol 115 (1) ◽  
pp. 30-36 ◽  
Author(s):  
Kanji Kasagi ◽  
Junji Konishi ◽  
Yasuhiro Iida ◽  
Yasutaka Tokuda ◽  
Keisuke Arai ◽  
...  
Keyword(s):  
Fold Increase ◽  
Graves Disease ◽  
Thyroid Cells ◽  
Thyroid Stimulating Antibodies ◽  
Silent Thyroiditis ◽  
Poorly Differentiated ◽  
Pre Treatment ◽  
Rat Thyroid ◽  
Serum Tsh ◽  
Bovine Tsh

Abstract. A sensitive, precise and practical assay for thyroid stimulating antibodies was developed in which poorly differentiated rat thyroid cells (FRTL-5) were exposed to crude immunoglobulin fractions precipitated from serum with 15% polyethylene glycol under hypotonic conditions. After the incubation at 37°C for 2 h, cAMP released into Hank's medium without NaCl was determined by radioimmunoassay. The removal of NaCl from the isotonic Hank's medium greatly enhanced cAMP production in response to both TSH and thyroid stimulating antibodies. The assay was sensitive enough to elicit an approximately 30-fold increase in cAMP at 10 mU/l bovine TSH. Thyroid stimulating activities measured using FRTL-5 cells significantly correlated with those measured using cultured porcine (r = 0.918, N = 72) or human (r = 0.830, N = 23) thyroid cells. Thyroid stimulating activities were detected in all of the 50 patients with hyperthyroid Graves' disease, the 14 patients with recurrent hyperthyroid Graves' disease, and the 25 patients with ophthalmic Graves' disease. Thyroid stimulating activity was also detected in some patients (9/24, 37.5%) with Hashimoto's thyroiditis whose serum TSH concentrations were higher than 30 mU/l. However, it was completely abolished by pre-treatment of the sera with anti-TSH antibodies. Although thyroid stimulating activities were detected in one of the patients with simple goitre (N = 10) and in one with thyroid cancer (N = 10), none of the patients with silent thyroiditis (N = 7), adenomatous goitre (N = 11), and thyroid adenoma (N = 9) were positive for thyroid stimulating antibodies.

scholarly journals Development of a Luminescent Bioassay for Thyroid Stimulating Antibodies

1999 ◽  
Vol 84 (1) ◽  
pp. 374-377 ◽  
Author(s):  
C. Evans ◽  
N. G. Morgenthaler ◽  
S. Lee ◽  
D. H. Llewellyn ◽  
R. Clifton-Bligh ◽  
...  
Keyword(s):  
Reference Range ◽  
Thyroid Autoimmunity ◽  
Light Output ◽  
Fold Increase ◽  
Graves Disease ◽  
Dependent Manner ◽  
Multinodular Goitre ◽  
Thyroid Stimulating Antibodies ◽  
Luminescent Assay ◽  
Dose Dependent

The hyperthyroidism of Graves Disease (GD) is due to thyroid stimulating antibodies (TSAb) which are thyrotropin (TSH) agonists. They are detected routinely by measuring their ability to inhibit TSH binding to the receptor (TBII), which does not reflect their true biological activity. Current bioassays which measure cAMP by RIA, are not suitable for routine use. We have developed a luminescent bioassay for TSAb, by introducing a cAMP responsive luciferase construct into CHO cells stably expressing the human TSH receptor (TSHR). Clone lulu1 displays dose dependent TSH response detectable from 10 μU/ml and maximal at 10 mU/ml when a >25 fold increase in light output is obtained. 34 euthyroid sera were tested to determine a reference range, with values >1.5 relative light units (R.L.U.) being considered positive. An international TSAb standard responded in a dose dependent manner with 10 mIU/ml giving an R.L.U. of >10. The assay was adapted to a 96 well format for automatic readout and 100 treated GD samples (50 TBII negative and 50 TBII positive) were tested, 73% being positive. In contrast only 4% of 79 control sera from individuals with Hashimoto’s, non-thyroid autoimmunity or multinodular goitre produced R.L.U. >1.5. When 44 of the GD sera were compared in a traditional salt-free bioassay, 61% were positive compared with 75% in the new luminescent assay. In conclusion, we have developed a luminescent bioassay for TSAb, using unfractionated serum which is capable of high throughput suitable for routine use.

scholarly journals Orbital fibrosis in a mouse model of Graves' disease induced by genetic immunization of thyrotropin receptor cDNA

2011 ◽  
Vol 210 (3) ◽  
pp. 369-377 ◽  
Author(s):  
Shuang-Xia Zhao ◽  
Shanli Tsui ◽  
Anthony Cheung ◽  
Raymond S Douglas ◽  
Terry J Smith ◽  
...  
Keyword(s):  
Close Association ◽  
Graves Disease ◽  
Genetic Immunization ◽  
Thyroid Stimulating Antibodies ◽  
Thyroid Glands ◽  
Orbital Tissue ◽  
A Subunit ◽  
Receptor Cdna

The TSH receptor (TSHR) is the critical target for antibody production in Graves' disease (GD). Insulin-like growth factor 1 receptor (IGF1R) has been proposed as a second autoantigen in complications of GD such as orbitopathy. We attempted to induce orbital tissue remodeling in mice undergoing immunizations with plasmids encoding TSHR and IGF1R delivered byin vivoskeletal muscle electroporation, a procedure known to give a sustained, long-term antibody response. FemaleBALB/cmice were challenged with TSHR A-subunit or IGF1Rα subunit plasmid by injection and electroporation. Mice challenged with TSHR A-subunit plasmid resulted in high frequency (75%) of hyperthyroidism and thyroid-stimulating antibodies. But strikingly, immunization with TSHR A-subunit plasmid also elicited antibody to IGF1Rα subunit. Mice challenged in the same manner with IGF1Rα subunit plasmid produced strong antibody responses to IGF1R, but did not undergo any changes in phenotype. Simultaneous challenge by double antigen immunization with the two plasmids in distant anatomical sites reduced the incidence of hyperthyroidism, potentially as a consequence of antigenic competition. Thyroid glands from the TSHR A-subunit plasmid-challenged group were enlarged with patchy microscopic infiltrates. Histological analysis of the orbital tissues demonstrated moderate connective tissue fibrosis and deposition of Masson's trichrome staining material. Our findings imply that immunization with TSHR A-subunit plasmid leads to generation of IGF1R antibodies, which together with thyroid-stimulating antibodies may precipitate remodeling of orbital tissue, raising our understanding of its close association with GD.

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