Optimization and Clinical Assessment of a Radioreceptor Assay for Thyrotropin-Binding Inhibitor Immunoglobulins

Konishi, Junji; Kasagi, Kanji; Iida, Yasuhiro; Kousaka, T; Murakami, Takashi; Arai, Keisuke; Tokuda, Yasutaka; Torizuka, K

doi:10.1507/endocrj1954.34.13

Cited by 25 publications

(8 citation statements)

References 18 publications

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“…Although the buffer of 10 mmol/1 TRIS-HC1, 50 mmol/1 NaCl was rec¬ ommended in TBII assay using thyroid mem¬ branes (5)(6)(7)12,14), our observation in the present study revealed that modified Hanks'(-) was the best of the four buffers tested and TBII activities were much higher in the modified hypotonie Hanks'(-) medium using cells. [I2:T]bTSH binding to FRTL-5 cells was much greater in the absence than in the presence of NaCl, in agreement with the observa¬ tion by others (10,17,18).…”

Section: Discussioncontrasting

confidence: 51%

“…TBII assay A radioreceptor TBII assay using detergent solubilized porcine thyroid membranes (TBII-pm) was carried out using a commercial kit developed by Southgate et al (7) with a minor modification (14). Results were expressed as follows: (1-specific [12TjTSH binding in the presence of a test sample/specific [12T]TSH binding in the presence of a pooled normal sample) x 100%.…”

Section: Preparation Of Serum Crude Immunoglobulin Fractionmentioning

confidence: 99%

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Simultaneous measurement of TSH-binding inhibitor immunoglobulin and thyroid stimulating autoantibody activities using cultured FRTL-5 cells in patients with untreated Graves' disease

Lü

Kasagi²,

Hidaka³

et al. 1990

Acta Endocrinologica

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A new and practical assay was developed using cultured FRTL-5 cells for simultaneous assessment of TS-ab and TSH-binding inhibitor immunoglobulin, allowing direct comparison of these two activities under the same conditions. Subsequent to the TS-ab assay in which extracellular cAMP concentration in Hanks' medium without NaCl was determined, [125I]bTSH in this medium was added to observe the ability of serum Ig to inhibit the binding of [125I]bTSH to FRTL-5 cells. We found a much higher specific binding of [125I]bTSH to FRTL-5 cells and a much greater inhibition of [125I]bTSH binding to the cells exposed to Graves' Ig in hypotonic NaCl-free than in NaCl containing Hanks' medium, indicating that the binding of both TSH and Graves' Ig to the TSH receptor was salt-sensitive. The inhibitory activity of [125I]bTSH binding to the cells was 0.2\m=+-\4.6%(mean \m=+-\sd) in 45 normals. Inter-assay coefficients of variation in two positive controls with the mean values of 18.0 and 65.8% were 15.8 and 16.5%, respectively. Among 46 patients with untreated hyperthyroidism owing to Graves' disease, 45 (97.8%) were positive for TS-ab; 35 (76.1%) and 40 (87.0%) were positive for TSH-binding inhibitor in Ig assays using FRTL-5 cells and solubilized porcine thyroid membranes, respectively. TS-ab activities correlated less closely with TSH-binding inhibitory activities determined using FRTL-5 cells (r=0.576, p<0.001) than with those determined using porcine thyroid membranes (r=0.745, p<0.001). The correlation between TSH\x=req-\ binding inhibitory activities determined by the two methods, although statistically significant (r=0.537, p<0.001) was rather rough. Some patients clearly demonstrated discrepant results, suggesting the existence of heterogeneous populations of TSH receptor antibodies.It is well known that thyrotropin receptor anti¬ bodies present in serum of patients with Graves' disease are responsible for the development of hy¬ perthyroidism. These antibodies are called thy¬ roid-stimulating autoantibody (TS-ab) or TSHbinding inhibitory immunoglobulin (TBII), ac¬ cording to the different methods employed in the assay systems. TS-ab is considered to stimulate thy¬ roid function through the adenylate cyclase system and its stimulating activity can be assessed by mea¬ suring the cAMP production using thyroid cells (1,2). The TS-ab assay can be made considerably more sensitive by incubating cultured thyroid cells in hypotonie medium (1), more rapid, and more reproducible by using cell strain 4). TBII, on the other hand, can inhibit the binding of [12T]TSH to its receptor in radioreceptor assay using crude or detergent solubilized thyroid mem¬ branes (5-7).Since the establishment by Ambesi-Impiombato et al. (8), FRTL-5 cells have frequently been used for studies on thyroid cell growth, differentiation and function, and a number of papers have been published using this cell line (reviewed in 9). In accordance with the biological response to TSH, such as cAMP production, iodine uptake and thymidine incorporation, specific and s...

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Section: Discussioncontrasting

confidence: 51%

Section: Preparation Of Serum Crude Immunoglobulin Fractionmentioning

confidence: 99%

Simultaneous measurement of TSH-binding inhibitor immunoglobulin and thyroid stimulating autoantibody activities using cultured FRTL-5 cells in patients with untreated Graves' disease

Lü

Kasagi²,

Hidaka³

et al. 1990

Acta Endocrinologica

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“…An impaired response of the thyroid to TSAb in ophthalmic Graves' disease, presumably owing to some de¬ structive change, has been postulated (Teng et al 1977) and is partly proven by the fact that some of the patients are hypothyroid (Christy et al 1977). As another possibility, some qualitative difference between the TSH receptor antibodies in ophthal¬ mic and thyrotoxic Graves' disease might be spe¬ culated, considering the reported lower incidence of TSH-binding inhibitor immunoglobulins in pa¬ tients with ophthalmic than in those with thyro¬ toxic Graves' disease (Teng et al 1977;Konishi et al 1987). However, the in vivo mechanism by which TSAb induce clinical manifestation of hyperthyroidism still remains to be clarified.…”

Section: Discussionmentioning

confidence: 99%

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

Kasagi

Konishi

Iida

et al. 1987

Acta Endocrinologica

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100

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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\s=deg\Cfor 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.

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“…The first one, which has been widely used in clinical practice for two decades, is a precipitation assay with solubilized porcine thyroid TSH-receptor [13,14]; the second is an improved system that uses test tubes coated with recombinant human TSH receptor [15]. TBII was expressed as % binding inhibition to facilitate comparison of the two systems.…”

Section: Methodsmentioning

confidence: 99%

Seasonal Variation in Relapse Rate of Graves' Disease after Thionamide Drug Treatment

et al. 2003

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Abstract. Objective: Controversy abounds on the issue of seasonal variation in new onset of Graves' disease, partly due to the difficulty of precisely dating the exact start of symptoms. To address the possible relationship between climatic changes and disease activity from a different perspective, we reviewed time of relapse during regular follow-up after successful drug treatment with thionamides. Design: Retrospective analysis of a case series in a university clinic. Patients and measurements: We consecutively registered patients who experienced re-emergence of hyperthyroidism between 1992 and 2001 after successful antithyroid drug therapy. Excluded were subjects with superimposing painless thyroiditis, in postpartum, on immunomodulatory drugs, or off thionamides prematurely on their own volition. Results: Fifty-two patients recurred 2 to 36 months after drug cessation. The frequency was higher in spring and summer (March to August) than in autumn and winter (September to February). With a new coated-tube radioreceptor assay, TSH binding inhibitor immunoglobulin activity was detected in sera from 87.5% of the reworsened patients. Conclusions: Graves' disease tends to relapse more frequently in spring and summer. Further clinical studies are warranted to clarify underlying mechanism (s) for this seasonal variation.

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Optimization and Clinical Assessment of a Radioreceptor Assay for Thyrotropin-Binding Inhibitor Immunoglobulins

Cited by 25 publications

References 18 publications

Simultaneous measurement of TSH-binding inhibitor immunoglobulin and thyroid stimulating autoantibody activities using cultured FRTL-5 cells in patients with untreated Graves' disease

Simultaneous measurement of TSH-binding inhibitor immunoglobulin and thyroid stimulating autoantibody activities using cultured FRTL-5 cells in patients with untreated Graves' disease

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

Seasonal Variation in Relapse Rate of Graves' Disease after Thionamide Drug Treatment

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