Changes of thyroid stimulating antibody and thyroid stimulating blocking antibody in treatment of anti-thyroid drugs and combining with levothyrocine to hyperthyroidism / 中华内分泌代谢杂志

Objective Changes of thyroid stimulating antibody(TSAb) and thyroid stimulating blocking antibody(TSBAb) in the treatment of anti-thyroid drugs(ATDs), and the effect of ATDs combining with levothyrocine(LT4) on TSAb and TSBAb were analyzed. Methods Using recombinant Trxfus. TSHRn protein and Trxfus. TSHRc protein as antigens, and TSH receptor antibody(TRAb)-N(TSAb binding hot spots), TRAb-C(TSBAb binding hot spots)in the serum of thyroid disease patients were measured with ELISA. The changes of TRAb-N, TRAb-C over 36 months in 117 TRAb-N positive Graves′ patients with hyperthyroidism were analyzed retrospectively. In the course of treatment, 41 cases as A group with ATDs and LT4 treatment, 76 cases as B group with only ATDs, The changes of TRAb-N and TRAb-C were observed in the two groups. Results (1)According to the change of TRAb-N, 117 TRAb-N positive Graves′ patients with hyperthyroidism were different. In group Ⅰ, 10 patients continued to have persistently positive TSAb and continued to have hyperthyroidism, remission rate 0%. In group Ⅱ, 17 patients showed complicated TRAb-N changes, 12 of 17 patients got relapse, 5 of 17 patients got remission, remission rate 29.4%. And in group Ⅲ, with TRAb-N dropping gradually, 15 of 89 patients got relapse, 74 of the 89 patients got remission, remission rate 83.1%. Three groups were significantly different with x2 test(P0.05). Conclusion TSAb and TSBAb can be used to document TRAb-function, which is significant for us to predict the changes of thyroid function. During ATDs treatment, the temporary early low-dose application of LT4 did not significantly affect TSAb and TSBAb.

Related factors in predicting relapse of Graves' disease after treatment with antithyroid drugs / 中华内分泌代谢杂志

Objective To explore the value of thyroid-stimulating antibody(TSAb) and degree of goiter in predicting the outcome of Graves'disease after antithyroid drug treatment. Methods Seventy-one patients with Graves'disease were given antithyroid drugs for (2. 8±1. 4)years and then followed up for(22±6.0)months.Finally,age,gender,thyroid function,TSAb and goiter size at the time of drug withdrawal were compared between the relapsed and relieved groups. TSAb was measured in all patients by using HEK-hTSHR cells. Results Eleven of 71 patients relapsed during the follow-up after drug withdrawal. The relapse rate (42. 9% ,6/14)in patients with positive TSAb was significantly higher than that (8.8%, 5/57) in patient with negative TSAb (X2 = 9.97, P<0.01). The relapse rates in patients with normal size thyroid, Ⅰ degree goiter,Ⅱ degree goiter were 6.25%, 12.2%,35.7% respectively. TSAb activity, positive rate and goiter size of the relapsed patients at the time of drug withdrawal were significantly higher than those of relieved patients (P<0.05 or P<0. 01). Conclusion TSAb activity and goiter size at the time of drug withdrawal are two effective prognostic markers of relapse in Graves' disease treated with antithyroid drugs.

Correlation between the Thyroid Associated Ophthalmopathy and Thyroid Function State

PURPOSE: To determine the prevalence of thyroid-associated ophthalmopathy (TAO) and the correlation between TAO and the thyroid function state. METHODS: 40 patients with TAO were selected among 2,000 patients of thyroid disease between September 1, 1995 and December 31, 2000, and classified by the Van Dyk's ''NOSPECS and RELIEF'' classification. T3, T4, TSH and Thyroid stimulating antibody (TSAb) levels were examined before and after treatment of TAO and thyroid disease. Relationship between TAO and thyroid function state was evaluated by using paired t-test and logistic regression test. RESULTS: The prevalence of TAO was about 2% in thyroid disease patients. Clinical manifestations of TAO were eyelid retraction (75%), exophthalmos (62.5%), soft tissue periorbital swelling (42.5%), diplopia and extraocular muscle restriction (20%), keratopathy (2.5%) and optic neuropathy (2.5%). Thyroid function state decided by T3, T4 and TSH level had no correlation with the improvement of TAO (p<0.05). But, TSAb level in improved TAO group (n=8) significantly decreased after treatment of TAO and thyroid disease (p<0.05). CONCLUSIONS: Thyroid function restoration may not help to improve the course of TAO. But, low TSAb level can be thought as a significant marker in improvement of TAO.

The signal pathways involved in the effects of thyroid stimulating antibody on the secretory function of thyrocytes / 中国实用内科杂志

Objective To investigate the signal pathways involved in the effects of thyroid stimulating antibody(TSAb)on the secretory function of cultured thyrocytes.Methods (1)With the method of enzyme-linked immunosorbent assay (ELISA),the PKA and PKC activities were observed.(2)Inhibitors or activators of protein kinases (PKA and PKC) were used to activate or block the signal pathways and the secretion of T3 measured by radioimmunoassay.Results (1)TSAb activated both PKA and PKC in thyrocytes in a dose-,time-dependent manner(.P.

A Case of Neonatal Graveses Disease / 대한소아내분비학회지

Neonatal Gaves disease is a relatively rare condition due to transplacental passage of Thyroid-stimulating antibody(TSAb) from a mother with active or inactive Graveses disease or autoimmune thyroiditis. A 11-day-old female newborn was referred to our department of pediatrics from a local clinic because of low level T4(3.55microg/dl) concurrent with high level TSH (501.74uIU/ml) on the 5th day neonatal metabolic screening. But, our repeated laboratory data showed very high serum T4(59.6microg/dl), T3(1,600ng/dl), suppressed TSH(0.43uIU/ml), and the presence of TSH receptor antibody. Her mother was treated with propylthiouracil(PTU) for Graves disease during pregnancy. Therefore, we thought it was a delayed-onset neonatal hyperthyroidism, because the fetal thyroid gland was initially suppressed by antithyroid drug taken during pregnancy. After initiating antithyroid drug therapy for the hyperthyroid nature, TSH levels became elevated again, while thyroid hormone levels decreased. Maternal and infant blood samples at the 23th day after birth were examined for serum autoantibodies directed towards the TSH receptor(Thyrotropin-binding inhibitory immunoglobulin:TBII, Thyroid-stimulating antibody:TSAb, Thyroid-stimulating blocking antibody:TSBAb) and high levels of TBII and TSAb were detected. About 2 months after birth, TBII and TSAb decreased within normal limit, and then we could stop antithyroid medication in safety. We report here a case of neonatal Graveses disease with very high level of T4 and T3, but firstly presented as hypothyroid nature on neonatal screening because of the maternally transferred antithyroid drug, PTU.

A Case of Neonatal Hyperthyroidism with Unilateral Ear Anomaly

Neonatal hyperthyroidism is a very rare disorder occurring typically in the offspring of patients with Graves' disease or chronic thyroiditis. It is caused by the transplacental passage of thyroid stimulating antibodies (TSAb) from the mother to the fetus. There has been few reports of neonatal hyperthyroidism associated with congenital anomalies. We experienced a case of neonatal hyperthyroidism with unilateral microtia and agenesis of external auditory canal in a female neonate born to a mother who had euthyroid but with a thyroid stimulating antibody. The patient was presented with unusual alertness, tachycardia, tachypnea, watery diarrhea, periorbital swelling and exophthalmos. Diagnosis was made by thyroid function tests and TSAb. She was treated with Lugol solution, PTU and propranolol. New she is 6 months old and in good condition with no symptoms of hyperthyroidism.

Difference of Thyroid Stimulating Antidody Activities Measured in Chinese Hamster Ovary Cells Stably Transfected with Human TSH Receptor and in FRTL-5 Cells in Graves Disease and Its Clinical Correlations / 대한내분비학회지

Background : Thyroid stimulating antibodies result in the development of hyperthyroidism and goiter in Graves disease. However, thyroid stimulating antibody activities do not correlate with the clinical features in many patients with Graves disease. The purpose of this study is to address this discrepancy between thyroid stimulating antibody activities and clinical features of Graves patients. Methods: We measured thyroid stimulating antibody activities simultaneously using human TSH receptor transfected Chinese hamster(hTSHR-CHO) cells and rat thyroid(FRTL-5) cells in 57 untreated patients with Graves disease, and compared their activities with clinical features including thyroid hormone levels. Results : The detection rate of thyroid stimulating antibody measured by hTSHR-CHO cells was 90% in 57 untreated Graves patients and it was higher than that measured by FRTL-5 cells. Thyroid stimulating antibody activity by hTSHR-CHO cells was significantly correlated with that by FRTL-5 cells(r=0.5, p<0.001), however, 18 of 57(32%) patients showed marked discrepancy of thyroid stimulating antibody activity between in hTSHR-CHO and FRTL-5 systems. Thyroid stimulating antibody activity measured by hTSHR-CHO cells was significantly correlated with serum total T3, free T4 levels, and goiter size but not 99mTc-thyroid uptake. On the other hand, thyroid stimulating antibody activity measured by FRTL-5 cells was significantly correlated with goiter size and 99mTc-thyroid uptake but not thyroid hormone levels. The difference between function and goiter size with respect to thyroid stimulating antibody measurement in two cells system is, nevertheless, particularly evident in the free T4/goiter ratio in patients with high hTSHR-CHO and low FRTL-5 cell assay values. Conclusion: These findings suggest that thyroid stimulating antibodies in Graves disease are heterogeneous population in terms of responses to different origin of cells. Further, thyroid stimulating antibody activities measured by FRTL-5 cells tend to correlate better with goiter size and Tc-thyroid uptake, whereas thyroid stimulating antibody activities measured by hTSH-CHO cells correlate better with thyroid hormone levels.

Thyroid Stimulating Antibody Assay with Chinese Hamster Ovary Cells Expressing Human Thyroid Stimulating Hormone (TSH) Receptor; Optimization of Assay Condition / 대한내분비학회지

We investigated the optimal condition of thyroid stimulating antibody(TSAb) assay using Chinese hamster ovary cells transfected with cDNA of human TSH receptor(TSHr-CHO) stably expressing functional TSH receptors. The extracellular cAMP responses of TSHr-CHO cells to the stimulation of bTSH or Graves' IgG were observed in three different incubation media. Stimulation indices of extracellular cAMP were higher when sucrose containing NaCl-free isotonic Hank's balanced salt solution(HBSS)(media A)was used as incubation media than those of NaCl-free hypotonic HBSS(media B) or those of NaCl containing isotonic HBSS(media C). The incubation of TSHr-CHO cells in media B caused marked increase in the basal cAMP level without concomittant fold-increase in the stimulated cAMP level at various doses of bTSH and Graves' IgG. Decreasing the stimulation indices of extracellular cAMP, use of media B failed to detect TSAb activities in two TSAb-positive Graves' IgG tested. In case of media C, extracellular cAMP responses are poor at 0.001 and 0.1U/L of bTSH and at all doses of Graves' IgG tested(0.5, 1, 5g/L). The incubation of TSHr-CHO cells in media B caused significant increase in the number of trypan blue-stained, nonviable cells(5.7+-1.5, 7.6+-1.9 and 8.5+-1.6% at 1, 2 and 3h of incubation, respectively; p<0.01) comparing to those incubated in media A or media C(about 2-3% in both media). Those decrease in the viability of TSHr-CHO cells when incubated in hypotonic incubation media may explain the decrease in the stimulation index of extracellular cAMP with the use of media B in contrast to the case of FRTL-5 cells. TSAb assay of 87 consecutive fresh Graves' patients with TSHr-CHO cells using media A detected TSAb activities in 90%(78 patients) of them, and moreover TSAb activities showed significant positive correlation with the pre-treatment serum T_3 and free T_4 levels of those patients. We conclude that TSAb assay with TSHr-CHO cells is a sensitive and physiologically relevant assay system to measure TSAb activities merely through measurements of extracellular cAMP provided that the cells are incubated in NaCl-free isotonic incubation media.

Assay of Thyrotropin Receptor Antibodies with Recombinant Human Thyrotropin Receptor Expressed on Chinese Hamster Ovary Cells / 대한내분비학회지

Thyroid stimulating antibody which results in the development of hyperthyroidism and goiter in Graves' patients used to be measured by using rat thyroid cells, FRTL-5. However, this assay has disadvantages: decreased sensitivity due to differences in species, and fastidious culture conditions for FRTL-5 cells. Thus, we recently created stably transfected Chinese hamster ovary(CHO) cells containing the human TSH receptor(hTSHR-CHO) and developed optimal conditions for the measurement of thyroid stimulating antibody using hTSHR-CHO cells. In this study, to evaluate the clinical relevance of thyroid stimulating antibody measurement using hTSHR-CHO cells, we measured thyroid stimulating antibody activities of IgGs from Graves' disease and other thyroid disease using hTSHR-CHO cells, and compared to those of thyroid stimulating antibody assays using FRTL-5 cells. 1) The cut off value of positive thyroid stimulating antibody activity measured in hTSHR-CHO cells was 145%(above the mean +2SD) which was lower than 165% in FRTL-5 cells. The intra-assay and inter-assay variances were 3.9% to 9.0% and 12.7% to 1.6%, respectively. 2) Thyroid stimulating antibody activity was detected in 90% of patients with untreated Graves' disease when patients initially presented. Further, in patients seen initially but already under therapy, 75% had positive values if they were hyperthyroid but only 43% had IgGs with activity if they were euthyroid. Patients in clinical remission after therapy showed positive values in 23% of cases. Only 2 of 25 patients with Hashimoto's thyroiditis showed weak thyroid stimulating antibody activity, none of 18 patients with nodular nontoxie goiter, 1 of 15 patients with primary myxedema, and 2 of 33 control patients with no thyroid disease. Thus, the detection frequency and specificity of the assay with hTSHR-CHO cells was excellent for this type bioassay.3) The detection frequency of thyroid stimulating antibody activity by hTSHR-CHO cells assay system(90%) was higher than that by FRTL-5 cells assay system(66%) in untreated Graves' patients. Those two activities were positively correlated with each other(r=0.52, p<0.001). However, some IgGs showed discrepancy of the thyroid stimulating antibody activity measured in hTSHR-CHO cells and in FRTL-5 cells; 56 of 87 patients were positive in both cells system, 8 of 87 were negative in both cells system, 1 of 87 was only positive in FRTL-5 cells and 22 of 87 were only positive in hTSHR-CHO cell system. Thus, 73%(22/30) of IgGs showing negative values of thyroid stimulating antibody activities in FRTL-5 cells were detected its activities in hTSHR-CHO cells system.In summary, thyroid stimulating antibody assay with hTSHR-CHO cells exhibited so excellent sensitivity and specificity that this technique should be used for clinical practice as well as basic research.

Negative correlation between the conversion of thyrotropin receptor-bound blocking type thyrotropin receptor antibody to the stimulating type by anti-human IgG antibodies and the biological activity of blocking type thyrotropin receptor antibody

It has been reported that receptor-bound blocking type TSH receptor antibody (TRAb) can be converted to the stimulating type by anti-human IgG antibodies. To evaluate the relationship between the conversion of receptor-bound blocking type TRAb to the stimulating type and the biological activity of blocking type TRAb, we compared converting activities of blocking type TRAb from 10 patients with primary nongoitrous hypothyroidism with both the doses of blocking type TRAb which show 50% inhibition of 125I-bTSH binding to the TSH receptor and those which show 50% inhibition of TSH-stimulated cAMP production in cultured rat thyroid cells (FRTL-5). The additions of anti-human IgG antibody to FRTL-5 cell-bound blocking IgGs resulted in the increase in cAMP production in a dose-dependent manner and the converting activities (percent increase of cAMP production) also depended on the doses of blocking IgGs. The converting activities were significantly correlated with the doses of blocking IgGs which showed 50% inhibition of 125I-bTSH binding to the TSH receptor (r = 0.71, p = 0.011). And these converting activities were also significantly correlated with the doses of blocking IgGs which showed 50% inhibition of TSH-stimulated cAMP increase (r = 0.81, p = 0.002), and were negatively correlated with thyroid stimulation blocking antibody activities (r = 0.58, p = 0.02). We have demonstrated that all cell-bound blocking type TRAb were converted to the stimulating type by anti-human IgG antibody and the degree of conversion was negatively correlated with the biological activity of blocking type TRAb.(ABSTRACT TRUNCATED AT 250 WORDS)

DETERMINATION OF THYROID-STIMULATING ANTIBODIES WITH CYCLIC AMP ACCUMULATION INDUCED BY UNFRACTIONATED SERUM USING CULTURED HUMAN THYRIOD CELLS / 中华内分泌代谢杂志

Thyroid-stimulating antibodies were determined with cyclic AMP accumulation induced by unfractionaled serum using cultured human thyroid cells. Thyroid-stimulating antibodies were found in 83.3% (25/30) of patients with Graves' disease without treatment, 43.8% (7/16) of patients with Graves' disease during antithyroid treatment, 4% (1/25) of normal people and none of patients with simple goitre or thyroid adenoma. The results indicate that determination of thyroid-stimulating antibodies using this method is sensitive and characteristic in patients with Graves' disease.

Effects of interferon-? on thyroglobulin and thyroid peroxidase gene expressions in FRTL5 cells induced by thyroid stimulating antibody / 中华内分泌代谢杂志

Objective To observe the effects of interferon ? (IFN?) on thyroglobulin (TG) and thyroid peroxidase (TPO) gene expressions in FRTL5 cells induced by thyroid stimulating antibody (TSAb). Methods TSAb crude fraction was extracted by polyethylene glycol 4000. Recombinant rat interferon ? (0, 1, 10, 10 2, 10 3 U/ml) was added to the FRTL5 cells induced by TSAb, and then expressions of TG, TPO mRNA were measured by Northern blot, the cell growth was measured by 〔 3H〕 thymidine incorporation. Results (1)TSAb increased TG, TPO gene expressions, and 〔 3H〕 thymidine incorporation. (2)Interferon ?inhibitedTSAb inducedTG, TPOgeneexpressionsand〔 3H〕 thymidine incorporation. Conclusion Interferon ? inhibits the growth and function of thyrocytes induced by TSAb, which suggests that interferon ? might regulate thyroid function in Graves' disease.

EVALUATION OF THE ASSAYS FOR THYROID-STIMULATING ANTIBODY MEASURING ~(125)I-UPTAKE AND cAMP RELEASE BY RAT THYROID CELLS / 中华内分泌代谢杂志

Thyroid-stimulating antibody (TSAb) was measured in 16 patients with active Graves' disease based on 125I-uptake-and cAMP release of rat thyroid cell strain FRTL5 according to Marcocci and Kasagi with modifications simultaneously. With the method of 126I-uptake, TSAb was found positive in 56% of the patients and the TSAb index was 1.19; with, the method of cAMP release, these two figures were 94% and 7.11 respectively. There were very significant differences for both. The method of cAMP release was sensitive and convenient:1) the cells may multiply continuously, the IgG-may be prepared easily,and the cAMP in the medium may be measured without extraction; 2) in 35 untreated and 42 treated patients with Graves' disease, the TSAb was positive in 91 % and 50% of the cases, and the TSAb activities were 1194% and 276% respectively, the differences were also very significant.

Effect of thyroid stimulating antibody on thyroid peroxidase activity in cultured thyrocytes / 中华内分泌代谢杂志

Objective To investigate the effects of thyroid stimulating antibody (TSAb) on the thyroid peroxidase (TPO) activity and its signal transduction pathways in cultured thyrocytes. Methods The TPO activity in primary cultured human thyrocytes was determined by amended guaiacol oxidation assay. At the same time, inhibitors of signal transduction were applied and the roles of cAMP/protein kinase A (PKA), phosphatidylinositol (PIP 2)/Ca 2+and tyrosine protein kinase (TPK) pathway on the TPO activity in cultured thyrocytes were observed. Results TSAb significantly enhanced the TPO activity in cultured thyrocytes, and the peak of TPO activity (360% of controls) reached with 4 mg/ml TSAb for 48 h. Addition of PKA inhibitor for blocking cAMP/PKA pathway decreased TPO activity by 79.2% (P

STUDY OF SERUM TRAb IN AUTOIMMUNE THYROID DISEASE WITH ABC- ELISA / 西安交通大学学报(医学版)

In this paper the investigation of using a new method-ABC-ELISA in assay of Autoimmune Thyroid Disease are presented. The sensitivity of ABC-ELISA is compared with that of standard ELISA; Its reliability is proven by the methods of detecting TSAb with FRTL-5. TRAb is detected by ABC-ELISA in 91% of untreated Graves'. TRAb is detected by Standard ELISA in 70% of untreated Graves'. The results of ABC-ELISA in 26 untreated Graves' are equal to that of the method of detecting TSAb with FRTL-5. Therefore, we consider that ABC-ELISA is a sensitive, reproducible, convenient method applicable to clinical practice.