Use of inactive beta-D-galactosidase for elimination of interference by anti-beta-D-galactosidase antibodies in immune complex transfer enzyme immunoassay for anti-thyroglobulin IgG in serum using beta-D-galactosidase from Escherichia coli as label

A novel enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-thyroglobulin IgG using beta-D-galactosidase from Escherichia coli as label was reported previously. This immunoassay was highly sensitive in demonstrating anti-thyroglobulin IgG not only in all patients with Graves' disease and chronic thyroiditis but also in a large proportion of healthy subjects. However, the detection of anti-thyroglobulin IgG at low levels in some serum samples was difficult, probably due to the presence of anti-beta-D-galactosidase antibodies. In the present study, the use of inactive beta-D-galactosidase was tested for elimination of interference by anti-beta-D-galactosidase antibodies. Preincubation of serum samples with excess of inactive beta-D-galactosidase resulted in sufficiently low backgrounds to detect low levels of anti-thyroglobulin IgG with little effect on the dose-response of anti-thyroglobulin IgG. As a result, it was revealed that anti-thyroglobulin IgG was present in almost all healthy subjects as well as all patients with Graves' disease and chronic thyroiditis.     

Measurement of anti-thyroglobulin IgG in urine of patients with autoimmune thyroid diseases by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay)

Anti-thyroglobulin IgG in urine of patients with Graves' disease and chronic thyroiditis and healthy subjects was measured by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay). Anti-thyroglobulin IgG in dialyzed urine was reacted simultaneously with 2,4-dinitrophenylated thyroglobulin and thyroglobulin-βT-D -galactosidase conjugate. The immune complex formed consisting of the three components was trapped onto polystyrene balls coated with (anti-2,4, dinitrophenyl group) IgG, eluted with ∈N-2,4-dinitrophenyl-L-lysine, and transferred onto polystyrene balls coated with (antihuman IgG γ-chain) IgG. β-D -Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. Anti-thyroglobulin IgG was detected in most of the patients, but not in most of the healthy subjects; levels of anti-thyroglobulin IgG in urine of the patients were well correlated to those in serum of the same patients. The measurement of anti-thyroglobulin IgG in urine by the immune complex transfer enzyme immunoassay was suggested to be useful as a diagnostic aid for autoimmune thyroid diseases. The conventional standard ELISA was not sufficiently sensitive for measuring anti-thyroglobulin IgG in urine. © 1993 Wiley-Liss, Inc.     

Molecular analysis of induced idiotypes associated with autoanti-thyroglobulin

Idiotype (Id) and autoanti-thyroglobulin were induced in different strains of mice by priming with anti-Id to monoclonal anti-thyroglobulins (D8 and G4) and challenged with a subimmunogenic dose of thyroglobulin (Tg). Both D8.Id and G4.Id were induced in CBA mice by priming with the appropriate anti-Id, but only priming with anti-D8.Id also induced an increase in anti-Tg. D8.Id was induced in other strains by the same schedule but it only appeared to be associated with anti-Tg in 129 and, to a lesser extent, BALB/c mice, both of which have the allotype Iga. The extent of the overlap between the D8 Id and the anti-Tg was estimated and shown to be greatest in the CBA strain from which the D8 clone was originally derived. Spectrotypic analysis of the induced Ids in CBA mice showed that some of the D8.Id, but none of the G4.Id, was identical to the original clonotype, implying that CBA mice normally have cells which can be induced to produce D8.Id-positive autoanti-Tg, which are normally weakly expressed or regulated. The observation that anti-D8.Id priming in some strains increased D8.Id-negative anti-Tg responses suggests that the D8.Id may also be associated with anti-Tg T-cells.     

Circulating Thyroid Antibodies in Congenital Hypothyroidism

ABSTRACT. The role of maternal thyroid antibodies in congenital hypotyroidism is controversial. We have analysed serum thyroid antibodies in patients and their mothers. In a bioassay, antibodies interacting with thyroid cells were analysed by measuring of TSH-stimulated CAMP production in a rat thyroid cell line, FRTLS. Serum antibodies against the TSH receptor, thyroid peroxidase and thyroglobulin were determined by radioreceptor assay and enzyme-linked immunosorbent assays. The bioassay was performed with IgG preparations from 89 mothers of children with congenital hypothyroidism. Analyses for TSH receptor antibodies and thyroid peroxidase/thyroglobulin antibodies were performed on 144 and 118 sera of newborn patients respectively. No evidence of an increased prevalence of thyroid antibodies was found on comparison with controls. One infant had transient neonatal hyperthyrotropinaemia because of TSH receptor blocking antibodies transferred from the mother. Our data indicate that, apart from transplacental transfer of TSH receptor antibodies, maternal immunoglobulins have a limited role in the aetiology of congenital thyroid dysfunction.     

The effect of vitamin D and selenomethionine on thyroid antibody titers,hypothalamic-pituitary-thyroid axis activity and thyroid function tests in men with Hashimoto’s thyroiditis: A pilot study

Background

Both selenium and vitamin D were found to reduce thyroid antibody titers in women with Hashimoto’s thyroiditis.

牛甲状腺球蛋白制备大鼠Graves病模型

目的：通过给大鼠注射牛甲状腺球蛋白,制备Graves病动物模型。方法：Wistar大鼠20只,分为对照组和模型组。模型组腹腔注射牛甲状腺球蛋白,每两周注射1次。对照组使用生理盐水代替。分别在第4周和第8周测定摄碘率、血清游离三碘甲状腺原氨酸（FT3）、血清游离甲状腺素（FT4）和促甲状腺素受体抗体（TRAB）。结果：第4周两组无明显差异。第8周两组出现明显差异,同时模型组中有6只大鼠达到甲状腺功能亢进（简称甲亢）标准。结论：使用牛甲状腺球蛋白制备大鼠Graves病模型是可行的。     

Heritability of levels of autoantibodies to thyroid antigens using the method of plotting regression of offspring on midparent (ROMP)

Only a few methods can be applied in a simple manner to estimate the genetic control of autoimmunity in humans. Here we examined the heritability of autoantibodies to two thyroid antigens; thyroglobulin (Tg) and thyroperoxidase (TPO, formerly known as thyroid microsomal antigen), using methods of regression of offspring on mid-parental values (ROMP). With the data sets available, affected and unaffected siblings were compared by this rapid screening method using results determined by hemagglutination (HA). The presence of both types of autoantibodies showed positive heritability in patients with Graves' thyrotoxicosis (TT), but it was not observed in chronic lymphocytic or Hashimoto's thyroiditis (CLT) patients. Since these assays have been extensively used over the years by most diagnostic and research laboratories, they should provide some insight as to which quantifiable parameters may be usefully accumulated to help select groups of patients and their families for further genetic study. ROMP may also be useful to determine the sequential appearance of different types of antibody in predicting disease onset in other family members, and in distinguishing maternal and paternal effects on imprinting. The method may be extended to study epitope spreading and other measures of disease progression.     

分化型甲状腺癌血液标志物的研究现状及进展

目的阐述近年来有关分化型甲状腺癌的血液标志物的研究现状及进展,以提高分化型甲状腺癌的临床诊治水平。方法对近年来国内外有关分化型甲状腺癌的血液标志物、液体活检等文献进行检索,对不同标志物进行归纳总结与分析。结果甲状腺球蛋白和甲状腺球蛋白抗体是分化型甲状腺癌目前最常用的血清学标志物,微小RNA、长链非编码RNA等血液标志物在分化型甲状腺癌的诊断、治疗和随访中的应用价值也不断被发现。结论因血液标志物具有特异度高、敏感度强且无创等优点,是未来协助提高甲状腺癌患者的诊断准确率、监测患者疾病进展及复发的有用指标。     

Genes and Environment as Predisposing Factors in Autoimmunity: Acceleration of Spontaneous Thyroiditis by Dietary Iodide in NOD.H2h4 Mice

In the field of autoimmune thyroiditis, NOD.H2^h4 mice have attracted significant and increasing attention since they not only develop spontaneous disease but they present thyroiditis with accelerated incidence and severity if they ingest iodide through their drinking water. This animal model highlights the interplay between genetic and dietary factors in the triggering of autoimmune disease and offers new opportunities to study immunoregulatory parameters influenced by both genes and environment. Here, we review experimental findings with this mouse model of thyroiditis.