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An improved method for the detection of thyroid hormone auto-antibodies (thaa) in serum
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L33 DEMONSTRATION OF HETEROGENEITY IN THYROTROPZN DISPLACING OR THYROID STIMULATING IMMTJNOGLOBULINS(IgGs) IN SERA OF PATIENTS WITH GRAVES' DISEASE BY FOUR DIFFERENT ASSAY METHODS. H. Uchimura, Y. Fukue, T. Mittsuhashi, K. Kubota, N. Kuzuya, H, Ikeda, Third Department of Internal MedQine, University of Tokyo, Tokyo, Japan. We have previously reported that thyroid stimularing activity by using porcine thyroid monolayer cultured cells was detected in al.1untreated patients with Graves' disease. Present study was intended to demonstrate the heterogeneity in IgGs from sera of untreated Graves' patients by measuring thyrotropin displacing activity (TDA) using human (h-) and porcine (p-) thyroidal TSH receptor, and thyroid stimulating activity (TSA) using -) thyroid monolayer cultured cells. TDA was ~~~~~-~~~~~~~~~~~ 1% I-bTSH, lmg IgG and human thyroidal membranes or solubilized porcine TSH receptor according to the method of Smith et al. TSA was assayed by using monolayer cells of human or porcine thyroids incubating with 5mg IgG for 2h followed by measurements of intracellular CAMP. IgGs of 22 untreated Graves' patients were prepared by Protein A Sepharose affinity chromatography. h-TSA was significantly correlated with h-TDA or p-TDA and no correlation between p-TSA and the others was observed. However p-TSA/h-TSA ratio was significantly correlated negatively with the others. When IgGs(blocking antibodies) of myxedema patients which showed neither p-TSA nor h-TSA were studied, very high h-TDA was paralleled with p-TDA. These results suggest that each of h-TDA, p-TDA and h-TSA might depict a common fraction of IgG class, albeit not identical, and that pTSA might indicate a quite heterogeneous fraction of IgG in patients with Graves' disease.
L34 AN IMPROVED METHOD FOR THE DETECTION OF THYROID HORMONE AUTO-ANTIBODIES (THAA) IN SERUM. S. Nakamura*, S. Sakata, T. Komaki, K. Kamikubo, K. Yasuda and K. Mlura. The 3rd Department Internal Medicine, Cifu Univ. School of Medicine, Gifu 500, Japan.
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Recently cases associated with THAA have been reported in patients with various thyroid diseases (such as Hashimoto’s thyroiditis and Graves’ In most of the reported cases, detection of serum THAA has been disease). performed under the condition in which intrinsic and/or therapeutic thyroid hormones could interfere with the binding of labelled thyroid hormones to In order to avoid such interference and to increase the serum THAA. sensitivity, we have developed and evaluated a new and simplified method for This include acidification of serum followed the detection of THAA in serum. by adsorption of liberated thyroid hormones onto dextran-coated charcoal and neutralization of the serum prior to conventional binding study. Sera from two patients with Hashimoto’s thyroiditis who had been demonstrated to have anti-T4 autoantibodies were used. Using our method, binding of ‘*‘l-T4 to serum THAA was the same in both cases whether a large amount of cold T4 had been added to the original sera or not. On the other hand, without the acid-treatment, addition of cold T4 inhibited the binding of 1251-T4 to serum It was concluded that serum THAA can be detected THAA in both cases. without the influence of circulating thyroid hormones when the present method is used.
L33 DEMONSTRATION OF HETEROGENEITY IN THYROTROPZN DISPLACING OR THYROID STIMULATING IMMTJNOGLOBULINS(IgGs) IN SERA OF PATIENTS WITH GRAVES' DISEASE BY FOUR DIFFERENT ASSAY METHODS. H. Uchimura, Y. Fukue, T. Mittsuhashi, K. Kubota, N. Kuzuya, H, Ikeda, Third Department of Internal MedQine, University of Tokyo, Tokyo, Japan. We have previously reported that thyroid stimularing activity by using porcine thyroid monolayer cultured cells was detected in al.1untreated patients with Graves' disease. Present study was intended to demonstrate the heterogeneity in IgGs from sera of untreated Graves' patients by measuring thyrotropin displacing activity (TDA) using human (h-) and porcine (p-) thyroidal TSH receptor, and thyroid stimulating activity (TSA) using -) thyroid monolayer cultured cells. TDA was ~~~~~-~~~~~~~~~~~ 1% I-bTSH, lmg IgG and human thyroidal membranes or solubilized porcine TSH receptor according to the method of Smith et al. TSA was assayed by using monolayer cells of human or porcine thyroids incubating with 5mg IgG for 2h followed by measurements of intracellular CAMP. IgGs of 22 untreated Graves' patients were prepared by Protein A Sepharose affinity chromatography. h-TSA was significantly correlated with h-TDA or p-TDA and no correlation between p-TSA and the others was observed. However p-TSA/h-TSA ratio was significantly correlated negatively with the others. When IgGs(blocking antibodies) of myxedema patients which showed neither p-TSA nor h-TSA were studied, very high h-TDA was paralleled with p-TDA. These results suggest that each of h-TDA, p-TDA and h-TSA might depict a common fraction of IgG class, albeit not identical, and that pTSA might indicate a quite heterogeneous fraction of IgG in patients with Graves' disease.
L34 AN IMPROVED METHOD FOR THE DETECTION OF THYROID HORMONE AUTO-ANTIBODIES (THAA) IN SERUM. S. Nakamura*, S. Sakata, T. Komaki, K. Kamikubo, K. Yasuda and K. Mlura. The 3rd Department Internal Medicine, Cifu Univ. School of Medicine, Gifu 500, Japan.
of
Recently cases associated with THAA have been reported in patients with various thyroid diseases (such as Hashimoto’s thyroiditis and Graves’ In most of the reported cases, detection of serum THAA has been disease). performed under the condition in which intrinsic and/or therapeutic thyroid hormones could interfere with the binding of labelled thyroid hormones to In order to avoid such interference and to increase the serum THAA. sensitivity, we have developed and evaluated a new and simplified method for This include acidification of serum followed the detection of THAA in serum. by adsorption of liberated thyroid hormones onto dextran-coated charcoal and neutralization of the serum prior to conventional binding study. Sera from two patients with Hashimoto’s thyroiditis who had been demonstrated to have anti-T4 autoantibodies were used. Using our method, binding of ‘*‘l-T4 to serum THAA was the same in both cases whether a large amount of cold T4 had been added to the original sera or not. On the other hand, without the acid-treatment, addition of cold T4 inhibited the binding of 1251-T4 to serum It was concluded that serum THAA can be detected THAA in both cases. without the influence of circulating thyroid hormones when the present method is used.