The cloning and sequencing of thyroid-stimulating hormone (TSH) receptor (TSHR), combined with advances in molecular techniques, have facilitated the understanding of the interaction of the TSHR antibodies (TSHRAbs) with the TSHR at the molecular level and have allowed the delineation of their clinical role. TSHRAbs in vivo are functionally heterogeneous; the stimulating TSHRAbs cause hyperthyroidism and diffuse goiter in patients with Graves' disease, whereas, the blocking TSHRAbs cause hypothyroidism in some patients with autoimmune hypothyroidism and are the cause of transient neonatal hypothyroidism. Measuring TSHRAbs has potential clinical implications in differential diagnosis of Graves' disease, predicting the outcome of Graves' disease after antithyroid drug treatment, and predicting the fetal/neonatal hyperthyroidism or neonatal hypothyroidism. The existence of epitope heterogeneity in a patient, i.e., of stimulating TSHRAbs with epitopes other than on the N-terminal region of the extracellular domain, is significantly associated with favorable long-term clinical response to antithyroid drug treatment. Measuring these subtypes for thyroid-stimulating antibody (TSAb) has potential clinical impli-cations, for example, in predicting responsiveness to treatment in untreated patients with Graves' disease.
Autoantibodies/*immunology ; Epitopes/immunology ; Graves Disease/diagnosis/immunology/therapy ; Humans ; Hyperthyroidism/diagnosis/*immunology/therapy ; Hypothyroidism/diagnosis/*immunology/therapy ; Immunoglobulins, Thyroid-Stimulating ; Receptors, Thyrotropin/*immunology

Animals ; Antibodies ; immunology ; therapeutic use ; Female ; Graves Disease ; immunology ; prevention & control ; Immune Tolerance ; immunology ; Mice ; Mice, Inbred BALB C ; Receptors, Thyrotropin ; immunology

PURPOSE: To investigate associations between serum thyroid stimulating hormone (TSH) receptor antibody (TRAb) levels and Graves' orbitopathy (GO) activity/severity in chronic-stage GO and compare the performance of two newly-developed TRAb assays (third-generation TSH-binding inhibition immunoglobulin [TBII] assay versus Mc4 thyroid-stimulating immunoglobulin [TSI] bioassay). METHODS: This study is a retrospective review of medical charts and blood tests from Korean GO patients who first visited the departments of ophthalmology and endocrinology, Yonsei University College of Medicine from January 2008 to December 2011, were diagnosed with GO and Graves' hyperthyroidism, and were followed up for > or =18 months. Third-generation M22-TBII and Mc4-TSI assays were performed in the chronic-inactive GO patients in whom euthyroidism status was restored. Patients' GO activity/severity clinical activity scores (CAS), and modified NOSPECS scores were examined for a correlation with TRAb assays. RESULTS: Fifty patients (mean age, 41.3 years; 41 females) were analyzed. The mean duration of Graves' hyperthyroidism symptom was 63 months (range, 18 to 401 months) and that of GO was 46 months (range, 18 to 240 months). All patients had been treated previously with anti-thyroid drugs for a median period of 52.3 months, and two patients underwent either radioiodine therapy or total thyroidectomy. Mean CAS and NOSPECS scores were 0.5 +/- 0.9 (standard deviation) and 4.8 +/- 3.1, respectively. Mean M22-TBII and Mc4-TSI values were 7.5 +/- 10.2 IL/L and 325.9 +/- 210.1 specimen-to-reference control ratio. TSI was significantly correlated with NOSPECS score (R = 0.479, p < 0.001); however, TBII was not associated with NOSPECS score (p = 0.097). Neither TSI nor TBII correlated with CAS (p > 0.05), because GO inflammatory activity subsided in the chronic stages of GO. CONCLUSIONS: In chronic-inactive GO after euthyroid restoration, GO activity score did not associate with serum levels of TRAb or TBII. However, levels of the functional antibody Mc4-TSI did correlate with GO severity. Therefore, the TSI bioassay is a clinically relevant measure of disease severity even in chronic inactive GO.
Adult ; Autoantibodies/*blood/immunology ; Chronic Disease ; Female ; Follow-Up Studies ; Graves Ophthalmopathy/*blood/diagnosis/immunology ; Humans ; Male ; Receptors, Thyrotropin/*blood/immunology ; Retrospective Studies ; Severity of Illness Index

We studied ninety cases of thyroid glands both histopathologically and by immunohistochemical methods in patients with Graves' disease using B and T cell markers to evaluate the role of lymphocytic subpopulation. Females were affected more frequently than males with a ratio of 6.5:1, and usually the females were younger than the males at the time of surgery. The heavier the lymphocytic infiltration, the higher was the percentage of germinal center formation or fibrosis. The degree of lymphocytic infiltration was also related to the titers of antithyroglobulin or antimicrosomal antibodies. T cells were mostly scattered individually or in small groups between the follicles; however, in the severely infiltrated group, the major pattern was in clusters. T8 positive cells were more abundant than T4 positive cells, and their distribution pattern was accordant with T11 positive cells. Immunoglobulin synthesizing B cells were positively stained in 47 of 94 cases tested and IgG was the most predominant. In the mild and moderate lymphocytic infiltration groups, IgM was mostly stained at the mantle zone or in the lymphoid cluster of the interfollicular stroma, whereas IgM positive cells were present exclusively in the germinal center of the severely infiltrated group. The results of our study indicate that the major lymphocyte subpopulation in Graves' disease is B lymphocytes, and the degree of T lymphocytic infiltration correlated better with titers of antimicrosomal antibody than antithyroglobulin.
Adolescent ; Adult ; Child ; Female ; Graves' Disease/immunology/*pathology ; Human ; Immunoglobulins/metabolism ; Lymphocytes/immunology/pathology ; Male ; Middle Age ; Support, Non-U.S. Gov't ; Thyroid Gland/immunology/pathology

The aim of study was to investigate the lymphocyte changes in peripheral blood of patients with Graves's disease accompanied by hematocytopenia and to explore its pathogenesis. The quantity and ratio of Th(2), Th(1), CD5(+) B, Bcl-2 level in 24 Graves's disease patients with hematocytopenia were detected by FACS, and 18 adults were selected as normal controls. The results indicated that the percentages of Th(1), Th(2), ratio of Th(2)/Th(1), CD5(+) B, Bcl-2 level in peripheral blood of the patients were (0.81 +/- 0.45)%, (6.83 +/- 3.02)%, (20.55 +/- 6.15)%, (20.89 +/- 1.62)%, (80.25 +/- 15.56)%, respectively, and were higher than those of normal controls [(0.39 +/- 0.24)% (P<0.05), (0.28 +/- 0.15)% (P<0.01), (0.52 +/- 0.12)% (P<0.01), (7.89 +/- 0.38)% (P<0.05), (36.49 +/- 6.79)% (P<0.05)]. It is concluded that the pathogenesis of Graves's disease with hematocytopenia may be related to unbalance of Th(1)/Th(2), increase of Th(2) inducing over-expression of CD5(+) B and Bcl-2 on B cell.
Adolescent ; Adult ; B-Lymphocytes ; immunology ; metabolism ; CD5 Antigens ; immunology ; Female ; Graves Disease ; complications ; immunology ; Humans ; Male ; Middle Aged ; Pancytopenia ; complications ; immunology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo explore the role of natural killer T (NK T) cells in the pathogenesis of Graves' disease.

METHODSNK T cell deficient mice and wild BALB/c mice were immunized with cells expressing TSH receptor once every two weeks 6 times. Two weeks after the final immunization, the mice were killed and serum thyroxine levels, anti-TSH receptor antibodies and thyroid pathological changes were examined.

RESULTSThe mean levels of TT(4) and TRAb in the immunized NK T cell deficient group were slightly elevated but significantly different from those of the non-immunized control group, while comparable to those in the immunized wild group. There were no significant changes of the activity levels of TSAb or TSBAb in the immunized NK T cell deficient mice compared to those in immunized wild control mice. Thyroids from immunized NK T cell deficient mice showed mild hypertrophy of some follicles as compared with non-immunized control mice. This change was comparable to immunized wild control mice.

CONCLUSIONNK T cells may not be involved in the pathogenesis of Graves' disease.

Animals ; CHO Cells ; Cricetinae ; Female ; Graves Disease ; etiology ; immunology ; Immunization ; Killer Cells, Natural ; physiology ; Mice ; Mice, Inbred BALB C ; Receptors, Thyrotropin ; immunology ; Thyroid Gland ; pathology

INTRODUCTIONIn a patient with hyperthyroidism, the detection of elevated thyroid hormone concentration with measurable thyroid-stimulating hormone (TSH) value poses considerable diagnostic difficulties.

CLINICAL PICTUREThis 38-year-old lady presented with clinical features of thyrotoxicosis. Her serum free thyroxine concentrations were unequivocally elevated [45 to 82 pmol/L (reference interval, 10 to 20 pmol/L)] but the serum TSH values were persistently within the reference interval [0.49 to 2.48 mIU/L (reference interval, 0.45 to 4.5 mIU/L)].

TREATMENTInvestigations excluded a TSH-secreting pituitary adenoma and a thyroid hormone resistance state and confirmed false elevation in serum TSH concentration due to assay interference from heterophile antibodies. The patient was treated with carbimazole for 18 months.

OUTCOMEThe heterophile antibody-mediated assay interference disappeared 10 months following the initiation of treatment with carbimazole, but returned when the patient relapsed. It disappeared again 2 months after the initiation of treatment.

CONCLUSIONSClinicians should be aware of the potential for interference in immunoassays, and suspect it whenever the test results seem inappropriate to the patient's clinical state. Misinterpretation of test values, arising as a result of assay interference, may lead to misdiagnosis, unnecessary and at times expensive investigations, delay in initiation of treatment and worst of all, the initiation of inappropriate treatment.

Adenoma ; diagnosis ; Adult ; Antibodies, Heterophile ; analysis ; immunology ; Diagnostic Errors ; Female ; Graves Disease ; diagnosis ; Humans ; Immunoassay ; Pituitary Neoplasms ; diagnosis ; Thyrotoxicosis ; blood ; diagnosis ; immunology ; Thyrotropin ; blood ; Thyroxine ; blood

OBJECTIVETo assess the association of tumour necrosis factor-α (TNF-α) gene polymorphisms at positions -863C/A, -857C/T, -238G/A and Graves disease (GD) susceptibility in Chinese Han population in Anhui region.

METHODSThe polymorphisms of TNF-α gene were determined by polymerase chain reaction with specific primers in 254 patients affected with GD and 212 healthy controls. Allelic and genotypic frequencies in GD group and normal controls as well as in different genders were compared. The allelic and genotypic frequencies for different thyroid stimulating hormone receptor antibody (TRAb) levels (TRAb > 12 U/L; ≤12 U/L) were also compared among patients with earlier onset GD.

RESULTS(1) The A allele at -863C/A locus in GD group (16.73%) was significantly greater than that of the control group (11.79%) (P< 0.05, OR = 1.503); the frequency of AA+CA genotype of -863C/A locus in GD group (32.68%) was significantly greater than that of control group (23.58%) (P< 0.05, OR = 1.573). There was no significant difference (P> 0.05) in the allelic and genotypic frequencies of -857C/T, -238G/A loci between the two groups. (2) There was no significant difference (P> 0.05) in the allelic and genotypic frequencies of -863C/A, -857C/T, -238G/A loci between patients of different genders. (3) There was no significant difference (P>0.05) in such frequencies between patients with earlier onset GD and different TRAb levels (TRAb > 12 U/L; ≤12 U/L).

CONCLUSION(1) The -863 A allele of TNF-α gene may contribute to the development of GD in Chinese Han population in Anhui, whilst -857C/T, -238G/A alleles may not. (2) There is no association between TNF-α gene -863C/A, -857C/T, -238G/A polymorphisms and development of GD in different genders. (3) There was no association between above polymorphisms and TRAb levels in patients with earlier onset GD.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Female ; Genetic Predisposition to Disease ; Graves Disease ; genetics ; immunology ; Humans ; Immunoglobulins, Thyroid-Stimulating ; genetics ; immunology ; Male ; Middle Aged ; Polymorphism, Genetic ; Receptors, Thyrotropin ; genetics ; immunology ; Tumor Necrosis Factor-alpha ; genetics ; immunology ; Young Adult

OBJECTIVETo evaluate the clinical validity of anti-thyroperoxidase antibody (anti-TPOAb) and anti-thyroglobulin antibody (anti-TgAb).

METHODSerum levels of anti-TPOAb and anti-TgAb were assayed using chemiluminescence immunoassay in 434 subjects, including 51 patients with Hashimoto's thyroiditis, 58 with Graves' disease, 68 with nodular goiter, 56 with thyroid adenoma and carcinoma, 56 with subacute thyroiditis, 65 with euthyroid non-thyroid endocrine disease, 35 with euthyroid non-thyroid autoimmune diseases, and 45 euthyroid controls.

RESULTSThe highest level and most positive results of serum anti-TgAb and anti-TPOAb were observed in patients with Hashimoto's thyroiditis (median 373 and 6 974 U/ml, positive rate 84.3% and 86.3%), followed by patients with Graves' disease (median 84 and 1 369 U/ml, positive rate 44.8% and 72.4%). Serum anti-TgAb and anti-TPOAb were also more common in patients with subacute thyroiditis and other autoimmune diseases than in the controls.

CONCLUSIONThe assay of serum anti-TPOAb and anti-TgAb by chemiluminescence immunoassy are useful in the differential diagnosis of autoimmune thyroid disease.

Adenoma ; blood ; Adolescent ; Adult ; Aged ; Autoantibodies ; blood ; Female ; Graves Disease ; blood ; Hashimoto Disease ; blood ; Humans ; Iodide Peroxidase ; immunology ; Male ; Middle Aged ; Thyroglobulin ; immunology ; Thyroid Gland ; immunology ; Thyroid Neoplasms ; blood ; Thyroiditis, Subacute ; blood

Serum thyroglobulin (Tg) is primarily used as a tumor marker to detect the recurrent or persistent disease in patients with differentiated thyroid carcinomas. Unfortunately, the serum Tg measurement is technically challenging and the thyroglobulin autoantibody (TgAb) interference remain the most serious problem limiting the clinical value of serum Tg. The direction and magnitude of the interference are related to the method and the concentration and affinity of the TgAb in the specimen. The objective of this study was to evaluate TgAb's interference with assay of Tg by electrochemiluminescent assay (ECLIA). The Tg and TgAb of 84 sera were measured by the ECLIA. Recovery tests were carried out in 3 groups. 3 different Tg calibrators, 50, 100, 200 ng/ml, respectively, were added into the sera of the first group. The sera of second group were doubly diluted 5 times, and meawhile the Tg and TgAb were measured after each dilution, and then 100 ng/ml Tg calibrator was added into the sera. The sera of third group were divided into different subgroups according to TgAb concentration and then 100 ng/ml Tg calibrators were added. Recovery rate (%) was calculated. Tg value decreased when TgAb concentration increased by ECLIA, TgAb value became lower when sera were diluted, Tg value increased. The added Tg calibrator had not significant influence on Tg value and Recovery rate. Recovery rate was lower when TgAb concentration increased. When sera were diluted, the recovery rate was increased. Tg value were underestimated by TgAb interference. After sera were diluted, Tg value became increasing by ECLIA. The added Tg calibrator had not significantly influence on Tg value and Recovery rate. When sera were diluted, the recovery rate was increased. TgAb concentration had significant influence on Recovery rate. Recovery test can not efficiently rectify Tg value when TgAb was positive.
Adolescent ; Adult ; Aged ; Autoantibodies ; blood ; immunology ; Child ; Electrochemical Techniques ; False Positive Reactions ; Female ; Graves Disease ; blood ; Hashimoto Disease ; blood ; Humans ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Thyroglobulin ; blood ; immunology ; Young Adult