A Receptor Assay for the Measurement of TSH Receptor Antibodies in Unextracted Serum

Southgate, Kay M.; Creagh, F. M.; Teece, Michelle; Kingswood, Chris; Smith, Bernard Rees

doi:10.1111/j.1365-2265.1984.tb00102.x

Cited by 189 publications

(82 citation statements)

References 12 publications

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“…In this method, 125 I-labeled bovine TSH compete with TRAb in serum samples to bind to purified porcine TSH receptors, followed by polyethylene glycol precipitation (13). After the completion of our study, new generations of TRAb assays have been developed (14,15), and it has been shown that a minor subset of sera from patients with Graves' disease are falsely negative using the assay employed in our study (15).…”

Section: Methodsmentioning

confidence: 99%

TSH-receptor autoimmunity in Graves' disease after therapy with anti-thyroid drugs, surgery, or radioiodine: a 5-year prospective randomized study

Laurberg

Wallin²,

Tallstedt³

et al. 2008

European Journal of Endocrinology

361

231

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Introduction: Autoimmunity against the TSH receptor is a key pathogenic element in Graves' disease. The autoimmune aberration may be modified by therapy of the hyperthyroidism. Objective: To compare the effects of the common types of therapy for Graves' hyperthyroidism on TSHreceptor autoimmunity. Methods: Patients with newly diagnosed Graves' hyperthyroidism aged 20-55 years were randomized to medical therapy, thyroid surgery, or radioiodine therapy (radioiodine was only given to patients R35 years of age). L-thyroxine (L-T 4 ) was added to therapy as appropriate to keep patients euthyroid. Anti-thyroid drugs were withdrawn after 18 months of therapy. TSH-receptor antibodies (TRAb) in serum were measured before and for 5 years after the initiation of therapy. Results: Medical therapy (nZ48) and surgery (nZ47) were followed by a gradual decrease in TRAb in serum, with the disappearance of TRAb in 70-80% of the patients after 18 months. Radioiodine therapy (nZ36) led to a 1-year long worsening of autoimmunity against the TSH receptor, and the number of patients entering remission of TSH-receptor autoimmunity with the disappearance of TRAb from serum during the following years was considerably lower than with the other types of therapy. Conclusion: The majority of patients with Graves' disease gradually enter remission of TSH-receptor autoimmunity during medical or after surgical therapy, with no difference between the types of therapy. Remission of TSH-receptor autoimmunity after radioiodine therapy is less common.

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Section: Methodsmentioning

confidence: 99%

TSH-receptor autoimmunity in Graves' disease after therapy with anti-thyroid drugs, surgery, or radioiodine: a 5-year prospective randomized study

Laurberg

Wallin²,

Tallstedt³

et al. 2008

European Journal of Endocrinology

361

231

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“…Most related studies have been performed using the first-generation TBII assay and yet second-generation assays are nowadays widely available. Instead of solubilized porcine receptors (14,15), these newer assays use recombinant TSHR coated on a solid-phase surface which provides a markedly improved sensitivity and specificity (16,17,18). To our knowledge, the threshold value indicating a risk for fetal or neonatal autoimmune hyperthyroidism with a second-generation TBII assay has only been investigated in one small study (19).…”

Section: Introductionmentioning

confidence: 99%

Predictive value of maternal second-generation thyroid-binding inhibitory immunoglobulin assay for neonatal autoimmune hyperthyroidism

Payrat

Chikh

Bossard

et al. 2014

European Journal of Endocrinology

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Context: Hyperthyroidism occurs in 1% of neonates born to mothers with active or past Graves' disease (GD). Current guidelines for the management of GD during pregnancy were based on studies conducted with first-generation thyroid-binding inhibitory immunoglobulin (TBII) assays. Objective: This retrospective study was conducted in order to specify the second-generation TBII threshold predictive of fetal and neonatal hyperthyroidism, and to identify other factors that may be helpful in predicting neonatal hyperthyroidism. Methods: We included 47 neonates born in the Lyon area to 42 mothers harboring measurable levels of TBII during pregnancy. TBII measurements were carried out in all mothers; bioassays were carried out in 20 cases. Results: Nine neonates were born with hyperthyroidism, including five with severe hyperthyroidism requiring treatment. Three neonates were born with hypothyroidism. All hyperthyroid neonates were born to mothers with TBII levels O5 IU/l in the second trimester (sensitivity, 100% and specificity, 43%). No mother with TSH receptor-stimulating antibodies (TSAb measured by bioassay) below 400% gave birth to a hyperthyroid neonate. Among mothers of hyperthyroid neonates, who required antithyroid drugs during pregnancy, none could stop treatment before delivery. Analysis of TBII evolution showed six unexpected cases of increasing TBII values during pregnancy. Conclusion: Maternal TBII value over 5 IU/l indicates a risk of neonatal hyperthyroidism. Among these mothers, a TSAb measurement contributes to identify more specifically those who require a close fetal thyroid ultrasound follow-up. These results should be confirmed in a larger series.

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“…TSH binding inhibition assays were carried out according to the method of Southgate et al (1984) using native human TSHR or recombinant human TSHR (expressed in CHO cells). Briefly, 50 µl detergent solubilised TSHR were preincubated with 50 µl sera, purified IgG or Fab fragment for 15 min at room temperature before the addition of 100 µl 125 I-TSH (30 000 c.p.m.)…”

Section: Tsh Binding Inhibition Assaysmentioning

confidence: 99%

Binding characteristics of antibodies to the TSH receptor

Oda

Sanders²,

Roberts³

et al. 1998

Journal of Molecular Endocrinology

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We have used fragments of the TSH receptor (TSHR) expressed in E. coli as glutathione S-transferase fusion proteins to produce rabbit polyclonal antibodies and a panel (n=5) of monoclonal antibodies to the extracellular fragment of the TSHR. The binding characteristics of the antibodies to linear, conformational, glycosylated and unglycosylated forms of the receptor in different assay systems have been investigated.The reactivity of these antibodies with the TSHR was assessed by Western blotting with both native and recombinant human TSHR expressed in CHO cells, immunoprecipitation of 35 S-labelled fulllength TSHR produced in an in vitro transcription/ translation system, immunoprecipitation of 125 I-TSH/TSHR complexes, inhibition of 125 I-TSH binding to the TSHR and fluorescence activated cell sorter (FACS) analysis of binding to CHO-K1 cells expressing the TSHR on their cell surface. Fab fragments of monoclonal antibodies were isolated, labelled with 125 I and used to determine the affinity constants of these antibodies with receptor, bound and free Fab being separated by polyethylene glycol (PEG) precipitation.Rabbit polyclonal and mouse monoclonal antibodies reacted with the TSHR in Western blotting and one monoclonal antibody (3C7) was able to inhibit 125 I-TSH binding to native human TSHR (74% inhibition), recombinant human TSHR (84% inhibition) and porcine TSHR (65% inhibition). Affinity constant values for TSHR monoclonal antibody Fab fragments calculated using Scatchard analysis were about 10 7 M 1 . Four out of five monoclonal antibodies reacted in FACS analysis with TSHR expressed on the surface of CHO-K1 cells. The FACS unreactive monoclonal (3C7) bound well to detergent solubilised TSH receptors and this emphasised the importance of using a combination of FACS analysis and radioactivelylabelled probes in analysis of the TSH receptor.The monoclonal antibodies produced in this study were found to be of relatively low affinity but proved useful for detection of the receptor by Western blotting and by FACS analysis.

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A Receptor Assay for the Measurement of TSH Receptor Antibodies in Unextracted Serum

Cited by 189 publications

References 12 publications

TSH-receptor autoimmunity in Graves' disease after therapy with anti-thyroid drugs, surgery, or radioiodine: a 5-year prospective randomized study

TSH-receptor autoimmunity in Graves' disease after therapy with anti-thyroid drugs, surgery, or radioiodine: a 5-year prospective randomized study

Predictive value of maternal second-generation thyroid-binding inhibitory immunoglobulin assay for neonatal autoimmune hyperthyroidism

Binding characteristics of antibodies to the TSH receptor

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