Studies on the Action of Thyroid Stimulation Blocking Antibody (TSBAb) on Thyroid Cell Membrane

Chen, Wen; Inui, Takehiro; Ochi, Yukio; Kajita, Yoshihiro

doi:10.1089/thy.1994.4.479

Cited by 9 publications

(8 citation statements)

References 26 publications

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“…Some sera were obtained from the Graves’ patients who became euthyroid with decreased TSAb activities during antithyroid drug therapy. Thyroid stimulation-blocking antibody (TSBAb)-positive hypothyroidism (during T 4 treatment) with high TSH-binding inhibitory immunoglobulin (TBII) activity were obtained from the thyroxine-treated patients with hypothyroidism [5]. …”

Section: Methodsmentioning

confidence: 99%

“…0.2 ml of these dissolved fractions were assayed in a total volume of 0.25 ml for 5 h at 37°C. cAMP released into the medium was determined as described previously [4, 5, 6, 7]. TS activity was expressed as the percentage increase of cAMP by test globulin compared to the mean values of the 10 control globulins (normal <150%).…”

Section: Methodsmentioning

confidence: 99%

“…TSBAb activity was determined by co-incubation of bTSH and PEG 12.5% PF from TSBAb-positive serum as described previously [5, 8]. All experiments were performed in duplicate or triplicate determinations.…”

Section: Methodsmentioning

confidence: 99%

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Clinical Usefulness of TSAb Assay with High Polyethylene Glycol Concentrations

et al. 1999

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We previously demonstrated the stimulatory effect of polyethylene glycol (PEG) on thyroid-stimulating antibody (TSAb)-IgG-stimulated cAMP production (thyroid stimulating (TS) index) in porcine thyroid cell (PTC) assay. In the present study the clinical usefulness of the practical method using high PEG concentrations was examined. TS activity using PEG 22.5% precipitated fraction (PF) was significantly higher compared to standard TSAb activity using 12.5% PF from TSAb-positive serum, but the maximum TS activity was observed with PEG 12.5% PF + 4% PEG or PEG 22.5% PF + 2% PEG. In all cases of untreated Graves’ patients, TSAb activity determined by PEG 22.5% PF was higher compared to standard TSAb activity using PEG 12.5% PF from test serum, but the highest TSAb activity was observed by PEG 12.5% PF + 4% PEG without increased cAMP production to normal serum. TSAb was positive in 85% (40/47), 98% (46/47) and 100% (47/47) of untreated Graves’ patients by the method of PEG 12.5% PF, PEG 22.5% PF and PEG 12.5% + 4% PEG, respectively. Increased TSAb activity by PEG 12.5% PF + 4% PEG method was also observed even if the standard TSAb activity using PEG 12.5% PF method was negative in the euthyroid states of Graves’ patients during antithyroid drug therapy. The stimulatory effect of PEG on TS activity was not found in other thyroidal diseases [thyroiditis chronica (with high serum TSH), thyroid stimulation-blocking antibody (TSBAb)-positive sera (with low serum TSH), adenomatous goiter, subacute thyroiditis, and thyroid cancer]. The stimulatory effect of 5% PEG on TS activity produced directly by small amounts of Graves’ serum (50 μl) was also found, although the sensitivity was lower than with PEG-precipitated IgG from 0.2 ml serum. The clinical usefulness of the sensitive TSAb assay using PEG-precipitated IgG or direct serum assay in the presence of high PEG concentrations was demonstrated.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Clinical Usefulness of TSAb Assay with High Polyethylene Glycol Concentrations

et al. 1999

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“…After the incubation the amounts of cAMP released into the medium were measured by Yamasa's RIA kit. TSAb assay and TSBAb assay using porcine thyroid cells were performed as previously described (11). T4 production activity using mouse bioassay was measured according to the method of McKenzie as previously described (12).…”

mentioning

confidence: 99%

Precise Determination of TSH Receptor Antibody Activity in Serum Containing Bovine TSH (bTSH) Binding Antibody by Absorption Using Denatured bTSH or Sheep FSH

Inui

Kouki²,

Okabe³

et al. 1996

Thyroid

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A previous report demonstrated that sera with bovine TSH (bTSH) binding antibody showed abnormally negative TSH receptor antibody (TRAb) activity in the standard TRAb assay method. The corrected TRAb activity calculated by the determination of the nonspecific binding of the labeled bTSH for each test serum [NSB(T)] resulted in positive TRAb activity. However, the precise calculation was difficult because NSB(T) level was significantly higher than the nonspecific binding in normal pool serum [NSB(N)] level. In the present experiment the determination of the TRAb activity was performed after absorption of bTSH binding antibodies by the heat-denatured bTSH to obtain more precise TRAb activity. In addition, absorption by sheep FSH (sFSH) was performed because almost all bTSH binding antibodies showed specific binding to the alpha-subunit of mammalian pituitary glycoprotein hormones in our previous study. Three days absorption of test serum using 1 mU of the heat-denatured bTSH (100 degrees C for 1 h) or 5 mU of sFSH was chosen as optimal because NSB(T) decreased remarkably to NSB(N) levels. The corrected TRAb determined after these absorptions decreased significantly compared to the corrected TRAb activity without the absorption. When the complete absorption of bTSH binding antibody was performed by the decrease of the NSB(T) level to the NSB(N) level, the TRAb activity determined by these two different absorptions was almost similar (difference was less than 10%). However, it was difficult to obtain the precise TRAb activity in the cases with extremely high bTSH binding antibody, because the NSB(T) level was higher than the NSB(N) level by the incomplete absorption of bTSH binding antibody.

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“…Then, cAMP production was assayed by Yamasa's RIA kit as described, previously (Chen et al, 1994). 25 ml for 5 hours at 37ЊC.…”

Section: Tsab and Tsbab Assay Using Porcine Thyroid Cells (Ptc)mentioning

confidence: 99%

Demonstration of fragments with thyroid stimulating activity from Thyroid stimulation blocking antibodies‐IgG molecules by papain digestion

Kouki

Inui

Yamashiro

et al. 1997

Clinical Endocrinology

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The conversion of Thyroid stimulation blocking antibodies activity to thyroid stimulating activity by papain digestion suggests that the inherent thyroid stimulating activity located in the Fab portion of the IgG molecule is unmasked by papain cleavage. We also suggest that the thyroid stimulating activity in the retarded fraction in the unbound fraction may be released from hydrolysis of the Fab portion of the IgG molecule.

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Studies on the Action of Thyroid Stimulation Blocking Antibody (TSBAb) on Thyroid Cell Membrane

Cited by 9 publications

References 26 publications

Clinical Usefulness of TSAb Assay with High Polyethylene Glycol Concentrations

Clinical Usefulness of TSAb Assay with High Polyethylene Glycol Concentrations

Precise Determination of TSH Receptor Antibody Activity in Serum Containing Bovine TSH (bTSH) Binding Antibody by Absorption Using Denatured bTSH or Sheep FSH

Demonstration of fragments with thyroid stimulating activity from Thyroid stimulation blocking antibodies‐IgG molecules by papain digestion

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