In vitro production of interferon-gamma by peripheral blood from patients with Graves'' disease, Hashimoto''s thyroiditis and rheumatoid arthritis.

The production of interferon-gamma (IFN-gamma) by peripheral blood mononuclear cells (PBMC), CD4 cells, or CD8 cells in response to interleukin-2 (IL-2) stimulation has been studied; the samples were obtained from 12 healthy control subjects, 19 patients with Graves' disease (10 hyperthyroid and nine euthyroid), 13 patients with Hashimoto's thyroiditis (four hypothyroid and nine euthyroid), and 15 patients with rheumatoid arthritis (11 active and four inactive). A dose of IL-2 (25 U/ml) was utilized to induce IFN-gamma by PBMC from all four groups. The incremental increase in IFN-gamma values (with IL-2 stimulation minus without stimulation) was significantly less in PBMC from patients with Graves' disease, Hashimoto's thyroiditis, and rheumatoid arthritis than that in PBMC from control subjects. The values from PBMC in patients with Graves' disease in a euthyroid state were below normal but greater than those from patients with Graves' disease in a hyperthyroid state. The incremental increase in IFN-gamma values from Graves' disease PBMC correlated with the serum TSH values (r = 0.622, P less than 0.01), but not with thyroid autoantibodies (anti-thyroid microsomal antibodies, anti-thyroid microsomal antibodies, nor TSH-binding inhibitory immunoglobulin activities). The incremental increase in IFN-gamma from PBMC from both control subjects and Graves' disease was correlated with that from CD4 cells (r = 0.711, P less than 0.01), but not with that from CD8 cells. The production of IFN-gamma in response to IL-2 from PBMC in Graves' disease correlated inversely with thyroid function, appearing to reflect the very effect of hyperthyroidism in this process. The precise explanation of these phenomena remains unclear. The decreased response of IFN-gamma to IL-2 stimulation by PBMC from patients with Graves' disease, Hashimoto's thyroiditis, and rheumatoid arthritis seems to be a non-specific phenomenon occurring in both organ specific autoimmune disease and systemic autoimmune disease. It may be due to a down-regulation in autoimmune disease of CD4 cells in response to IL-2, a decreased level of IL-2 cellular receptors or a decreased receptor affinity, associated increased soluble IL-2 receptors, or a defect of the intra-CD4 cellular IL-2 signal to produce or release IFN-gamma in the conditions studied.     

The Association of HLA with Autoimmune Thyroid Disease in Newfoundland. The Influence of HLA Homozygosity in Graves' Disease

Forty-seven patients with Graves' disease, 73 with thyroiditis and 128 controls drawn from the same geographical area of Newfoundland were HLA typed. The frequency of HLA-B8 was significantly increased in the Graves' disease patients when compared to the control group giving a relative risk of 3.9. There were no significant HLA differences between the thyroiditis and control groups.
Homozygosity for the HLA haplotype, which is common in this island population, was more common in Graves' disease patients (12.8%) than in controls (5.5%) but did not reach statistical significance in this sample. Homozygosity was due in five of the six cases to either an A1;B8 haplotype or an A2;B8 haplotype. This contrasted with an apparently random assortment of haplotypes in the control and thyroiditis groups. Calculations suggest that homozygosity for a B8 haplotype confers an additional risk over heterozygosity for B8 of about 3.5 fold; however, homozygosity had no observable influence on the severity of the disease.
These results strengthen the idea that B8, or an allele in linkage disequilibrium with it, determines in part the susceptibility of an individual to developing Graves' disease.     

格雷夫斯病患者体内抗氧化状态及氧化损伤的研究

目的:观察格雷夫斯病(GD)患者体内抗氧化状态及生物大分子的氧化损伤情况。方法: 检测GD初发组和治疗组各31例患者血浆总抗氧化能力(TAC)、超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-Px)活性及血浆丙二醛(MDA)和巯基(SH)含量,与31例正常对照比较;采用单细胞凝胶电泳法(SCGE)检测各组外周血单个核细胞(PBMC)的DNA损伤情况(用彗星率表示)。结果: GD初发组血浆TAC、SOD、GSH-Px活性及SH含量明显低于对照组(P＜0.05,P＜0.01),MDA含量及彗星率明显高于对照组(P＜0.01);GD治疗组各指标明显轻于初发组(P＜0.05,P＜0.01),但仍未达到对照水平。GD初发组及治疗组患者PBMC彗星率与TAC呈负相关,与MDA含量呈正相关。 结论:GD患者体内存在氧化应激及脂质、蛋白质和DNA的氧化损伤,生物大分子的氧化损伤可能参与GD的发生和发展。     

Assays of TSH-receptor antibodies in 576 patients with various thyroid disorders: their incidence, significance and clinical usefulness

The incidence and the significance of TSH-receptor antibodies in Graves' disease and in various thyroid disorders have been evaluated. TSH-binding inhibiting antibodies (TBIAb) and thyroid stimulating antibodies (TSAb) were detected in a large proportion of Graves' disease patients (TBIAb in 68.8% and TSAb in 77.8%), in a small number of patients with idiopathic myxoedema or Hashimoto's thyroiditis, and were not detected in patients with endemic euthyroid goitre, differentiated thyroid carcinoma and toxic adenoma. Furthermore, TSH-receptor antibodies were present in some patients with toxic multinodular goitre (TBIAb in 12.7% and TSAb in 15.9%). When TSH-receptor and other thyroid autoantibodies were compared, it was found that 13 of the 15 Graves' patients with negative tests for thyroglobulin and thyroid microsomal antibodies were positive for TSH-receptor antibodies. On the other hand, 9 of the 11 patients with toxic multinodular goitre who had positive TSH-receptor antibody tests, also had serum thyroglobulin and/or thyroid microsomal antibodies. No significant differences in the prevalence of TSH-receptor antibodies were found in Graves' patients irrespective of the presence of ophthalmopathy or pretibial myxoedema. Elevated TBIAb activity at the end of anti-thyroid drug treatment was found in 52.9% of Graves' patients who subsequently relapsed, while in Graves' patients in remission TBIAb was always negative. TSH-receptor antibody results were not predictive of the outcome of radioiodine treatment in Graves' disease. Finally no correlation could be found between TBIAb and TSAb in Graves' disease and Hashimoto's thyroiditis. In conclusion: the high incidence of TSH-receptor antibodies in Graves' disease confirms their pathogenetic role in the development of hyperthyroidism; TSH-receptor antibodies in Graves' disease are not significantly associated with the presence of ophthalmopathy or pretibial myxoedema; TSH-receptor antibody assays may be useful for the diagnosis of Graves' disease in the absence of other signs of autoimmunity. TBIAb seems to be a good predictor of relapse in Graves' patients treated with anti-thyroid drugs; a fraction of toxic multinodular goitre could be a nodular variant of Graves' disease.     

HL-A antigens in Graves' disease and Hashimoto's thyroiditis.

An increased frequency of HL-A 1 and HL-A 8 was found in patients with Graves' disease and Hashimoto's thyroiditis, whem compared to a control group from the same geographic area, and drawn from a pool of subjects attending one diagnostic centre. Furthermore, an increased incidence of W15 and W17 was found in Hashimoto's thyroiditis; however, these were not detected in any patient with Graves' disease.     

Increase of peripheral B lymphocytes in Graves' disease.

Peripheral T and B lymphocytes were examined in autoimmune thyroid diseases. The percentages of T and B lymphocytes were calculated from the proportions of E and EAC rosette-forming cells and peroxidase-positive cells determined by micromethods. In thyrotoxic Graves' disease, the percentage of T cells was significantly lower, and the percentage of B cells was higher than in normal controls. The absolute count of B lymphocytes was also markedly increased. The serum levels of thyroid hormones showed a significant correlation with the percentage of B cells and an inverse correlation with that of T cells in untreated cases of Graves' disease. Similar abnormalities of lymphocyte subpopulations were observed in patients with thyrotoxic Graves' disease under drug therapy, but the proportions and absolute counts of T and B lymphocytes were normal in euthyroid patients with Graves' disease, either under drug therapy or in remission. No abnormalities in T and B cells were found in Hashimoto's disease. The data indicate that the main feature of the abnormality of the lymphocyte subpopulations in thyrotoxic Graves' disease is an increase of B lymphocytes. The reasons for the discrepancy between our results and those of earlier reports and for the B cell abnormality in Graves' disease are discussed.     

Sequential psychological testing during the course of autoimmune hyperthyroidism

Summary Although the psychological disturbances accompanying Graves' disease are well known, the time required for normalisation of these disturbances during antithyroid drug treatment is not known. Therefore sequential psychological testing during the course of Graves' disease was done. There are also contradictory results concerning the possible correlation of neurophysiological and psychological test results during the course of Graves' disease with thyroid hormone values. Finally, psychological disturbances have been proposed as possible etiologic factors in Graves' disease.In our study, a significant decrease in anxiety and irritability could be observed at the time euthyroidism was achieved. Self-evaluations of depressivity, activity, exhaustion, well-being, extraversion, introversion, and the ability to concentrate changed 1 or 2 months after euthyroidism was induced. Similar test results could be observed after induction of euthyroidism by antithyroid drugs and subtotal thyroid resection. Therefore the mode of therapy does not seem to influence the course of normalisation of psychological parameters.In contrast to other investigations there was hardly any correlation between thyroid hormone values and psychological test results or the ability to concentrate. Nontheless, patients with Graves' disease showing high scores for depression and anxiety exhibit abnormal peripheral helper/suppressor T-lymphocyte relations. Furthermore, patients suffering from Graves' disease tend to be more anxious than controls. It remains to be determined whether an increased susceptibility to psychological disturbances has led to these alterations of lymphocyte subsets in Graves' disease patients with severe depression and anxiety.Supported by SFB 258     

HLA class II associations in juvenile Graves' disease: indication of a strong protective role of the DRBl*0701, DQAl*0201 haplotype

Previous studies have shown that HLA-DRB1*0301 and DQA1*0501 are associated with susceptibility to Graves' disease. Ninety Danish patients with early onset of Graves' disease and 102–192 controls were analyzed for HLA-DR and -DQ to investigate if the same associations exist in the juvenile form of Graves' disease. Both DRB1*0301 and DQA1*0501 were highly significantly increased in the patients with relative risks of 8.0 and 4.6, which are higher than those seen in adults. Stratification showed that DRB1*0301 is more strongly associated than DQA1*0501. Surprisingly, the DRB1*0701, DQA1*0201 haplotype was completely absent from this group of patients, indicating a strong protective role of this haplotype in juvenile Graves' disease.     

The association of HLA -A, -B,and -DRB1 genotypes with Graves' disease in Taiwanese people

Graves' disease has been associated with different human leukocyte antigen (HLA) genes in different races. To evaluate the association of HLA type in Taiwanese with Graves' disease, the HLA-A, -B, and -DRB1 alleles in a total of 236 Taiwanese adults with Graves' disease and 533 racially matched normal control subjects were examined using the PCR-SSOP (sequence specific oligonucleotide probe) technique. The prevalence of HLA-A*0207, -B*2704, -B*4601, and -DRB1*0901 among patients with Graves' disease was found to be increased, with odds ratios (OR) of 2.21, 3.82, 1.76 and 1.62, respectively. However, after correction for multiple comparisons, the relative risk of HLA-A*0207 susceptibility to Graves' disease remained statistically significant and the haplotype HLA-A*3303 -B*5801 -DRB1*0301 had a significantly protective effect. None of the other 2- or 3-locus haplotypes showed any significantly increased risk. Although HLA-DRB1*1405 showed an increased relative risk in patients with GO (Graves' opthalmopathy) (OR 4.61) when compared with patients without GO, the relative risk after adjusting for the number of comparisons was not significant. Taiwanese patients with Graves' disease have HLA-associated susceptibility genes which are similar to those found in Chinese patients in Hong Kong and Singapore. However, the finding in this study of a higher frequency of HLA-A*0207 in Taiwanese with Graves' disease has not been documented in any other ethnic group.     

Thyroid-stimulating antibody (TSAb) detected in sera of Graves'' patients using human thyroid cell cultures

As a homologous system is required to evaluate the effect of thyroid-stimulating antibody (TSAb) present in the serum of Graves' patients, primary cultures obtained from normal human thyroid gland have been used and the stimulatory effect measured as an increase of cAMP intracellular levels.

Monolayer cell cultures were stimulated by IgG purified from sera of Graves' patients or control subjects and compared to the effect of bovine TSH. Bovine TSH produced a dose-dependent increase in cAMP intracellular levels between 0·05 mU and 2·5 mU/ml, reaching a maximal value after 30 min with higher doses. While normal IgG had no effect, IgG prepared from untreated patients with frank Graves' disease elicited a significant increase in cAMP accumulation at a concentration between 0·05 and 0·5 mg/ml within 60 min in thirteen out of fourteen patients. A longer incubation period showed no further increase in cAMP values, even if in one case a higher concentration (5·0 mg/ml) of Graves' IgG had a delayed response. When the cAMP intracellular level modifications produced by Graves' IgG preparations in thyroid cell cultures were compared to those evoked in thyroid slices, an identical percentage (93%) of positive cases was obtained, without a coincidence of negative cases. Using thyroid slices the cAMP intracellular increase above basal levels was higher, if considered as a percentage, but in cultured cells a very low IgG concentration was sufficient to detect the presence of TSAb. No correlation between the two assays was found.

In conclusion, normal human cultured thyroid cells appeared to be a more suitable substrate when compared to human thyroid slices for detecting the presence of TSAb in Graves' disease and for studying its effect on thyroid cells. However, a 100% TSAb positivity was present in our Graves' patient series only when both assays were used.

     

TSH受体抗体测定的临床意义

目的:探讨血清TRAb的测定变化对G raves甲亢的临床意义。方法:应用放射免疫受体分析法(RRA)对302例甲状腺疾病患者及52例正常健康人的血清TRAb的值进行比较。结果:甲亢组TRAb的测定值阳性率为86.3%;甲亢缓解组TRAb的测定值阳性率为74.5%;甲亢治愈组TRAb的测定值阳性率为32.1%;甲亢复发组TRAb的测定值阳性率为90.3%;单纯性甲状腺肿组TRAb的测定值阳性率为0;甲瘤组TRAb的测定值阳性率为0;甲亢组、甲亢缓解组、甲亢治愈组、甲亢复发组与正常对照组相比有显著性差异(P<0.01);单纯性甲状腺肿组、甲瘤组与正常对照组相比无显著性差异(P>0.05)。结论:TRAb的测定对G raves甲亢的治疗具有重要的参考价值。     

Bereavement as an antecedent factor in thyrotoxicosis of childhood: four case studies with survey of possible metabolic pathways.

Various organic and psychosomatic factors have been postulated over the years as etiologic events antedating the onset of Graves' disease. In some patients psychological events have appeared to be important in the evocation of symptoms. Although examples of the latter have been described in adults for many years, there is little published on this phenomenon in children. The present study delineates findings in two boys and two girls with an age range of 8 to 14 years. Separating experiences appeared to be related to the onset or relapse of Graves' disease in these particular cases. In three of the patients the trigger event was represented by bereavement after death of a close relative; in the fourth case the boy's loss was enforced and traumatic separation from his mother figure. In all these children depression was the common response to loss. The observed relationship between the affective disturbance and Graves' disease is compatible with one or more hypothetical models. One such pathway, via depletion of brain monoamines associated with the state of depression, could cause an activation of the hypothalamic-pituitary-adrenal axis with resultant suppression of immune surveillance. This could permit the formation of thyroid-stimulating immunoglobulins (TSI) and hence Graves' disease in genetically susceptible (HLA B-8) persons.     

Human lymphocyte antigens (HLA) and Graves' disease in Turkey

To evaluate the association of HLA types with Turkish patients with Graves' disease, HLA typing, clinical findings, and thyroid antibodies were correlated. The HLA types, clinical findings (ophthalmopathy and age at onset), and thyroid stimulating hormone (TSH) receptor (TRAb) and antithyroid microsomal antibodies (MAb) were analyzed. Seventy Turkish patients with Graves' disease and 306 control subjects were assessed. Serological HLA typing was performed in HLA A, B, C, DR, and DQ loci. There was a significantly increased prevalence of HLA B8, B49, DR3, DR4, and DR10 in Graves' disease. The association of Graves' disease with HLA DR3 was found to be less strong than previously described. The HLA DR4 antigen may contribute to the predisposition of Graves' disease in Turkey. The results suggest that HLA B7, B13, DR7, DQw2, and DQw3 may confer a protective effect for Graves' disease in Turkey. Patients carrying HLA B12, B18, and B44 haplotypes had a tendency to develop the disease at a later age. The difference from the other studies may be the result of the selection of the controls; in part, of the variability in serological typing reagents; and, also, of the rather weak HLA associations with the disease.This study was presented in part at the Annual Meeting of the National Endocrinology and Diabetes Association, Bursa, Turkey, May 25–28, 1992.     

Graves病患者甲状腺和外周静脉血中甲状腺特异性抗体的对比研究

本文对比研究１１例Ｇｒａｖｅｓ病（ＧＤ）和９例非ＧＤ患者甲状腺静脉血（ＴＶＢ）和外周静脉血（ＰＶＢ）中甲状腺刺激抗体（ＴＳＡｂ）、甲状腺球蛋白抗体（ＴＧＡｂ）和甲状腺过氧化物酶抗体（ＴＰＯＡｂ）的活性和Ｔ３、Ｔ４浓度。结果显示：（１）抗体阳性的ＧＤ患者，其ＴＶＢ中ＴＳＡｂ、ＴＧＡｂ和ＴＰＯＡｂ水平均显著高于ＰＶＥ的，ＰＶＢ和ＴＶＢ的ＴＳＡｂ活性呈显著正相关，这提示甲状腺本身是甲状腺特异性抗体产生的主要部位；（２）ＧＤ组和非ＧＤ组ＴＶＢ和ＰＶＢ的血清Ｔ３、Ｔ４不形成浓度梯度；（３）ＴＳＡｂ、ＴＧＡｂ和ＴＰＯＡｂ活性及其在ＴＶＢ和ＰＶＢ之间的活性梯度，与ＴＶＢ和ＰＶＢ中Ｔ３、Ｔ４浓度均无相关关系。     

Sera but not immunoglobulins of ophthalmic Graves' patients stimulate human embryonal fibroblasts' biosynthetic activity in culture

The retroocular connective tissue changes in the ophthalmopathy of Graves' disease are known, however, the mechanism which leads to the increase in fibroblast number and activity is poorly understood. Using human embryonal fibroblast monolayers, fibroblast biosynthetic activity in the presence of sera, immunoglobulin deprived sera or immunoglobulins of Graves' patients with and without ophthalmopathy has been measured. Both sera and immunoglobulin deprived sera of the ophthalmic Graves' patients caused a marked increase of protein synthesis and a moderate increase of the sulphated glycosaminoglycan synthesis of fibroblasts. The same stimulatory effect was not found when immunoglobulin in fetal calf serum was used instead of sera, though anti-fibroblast IgG-s were present both in the sera and separated immunoglobulin fractions, as it has been demonstrated in an ELISA system. We conclude that the sera of ophthalmic Graves' patients contains a factor which stimulates human embryonal fibroblasts' biosynthetic activity in culture; this factor is not an immunoglobulin. The system described here seems to be suitable for studying the accompanying connective tissue changes in Graves' disease.     

Immunological features of sporadic multinodular goiter

Summary The origin of sporadic multinodular goiter is still uncertain. To obtain information on a number of unexplored immunological features, the distribution and characterization of T, B, and natural killer lymphocyte subsets were studied in the peripheral blood of 15 patients with multinodular goiter; 8 patients with Graves' disease (for reference purposes with a well-characterized autoimmune disease) and 29 age- and sex-matched healthy controls, combining double-staining immunofluorescence technique with monoclonal antibodies and flow cytometry. Although in both thyroid diseases increased CD3⁺ HLA-DR⁺ activated T cells (P < 0.01) were detected, in Graves' disease this was associated with decreased numbers of CD8⁺ cells (P<0.05) and an increased CD4/CD8 ratio (P < 0.01). These abnormalities were absent in multinodular goiter, which displayed increased CD8⁺ CD57⁺ cytotoxic/suppressor cells (P < 0.01). There was an increase in the percentage of natural killer cells expressing CD16 and CD57 antigens in multinodular goiter but not in Graves' disease. The B-cell associated antigens CD 19 and CD19⁺ CD5⁺ were significantly increased in Graves' disease (P < 0.01), while the multinodular goiter patients exhibited only an increased number of B cells coexpressing the CD5 antigen (CD19⁺ CD5⁺), which was unrelated to the titers of antimicrosomal and antithyroglobulin autoantibodies. Our results point to the presence of several abnormalities of peripheral T, B, and natural killer lymphocytes in sporadic multinodular goiter, with a distribution pattern quite different from that observed in Graves' disease. These results support the notion that, in contrast to Graves' disease, in sporadic multinodular goiter different suppressor and/or cytotoxic mechanisms are set up by the immune system, reflecting either pathogenic mechanisms of the disease or an immune response to pathological growing tissue.Abbreviations FITC fluorescein isothiocyanate - NK natural killer - PE phycoerythrin - T₃ triiodothyronine - T₄ thyroxine - TBII thyroid binding inhibitory immunoglobulin - TSH thyroid-stimulating hormone     

Secretor status and infection in patients with Graves' disease

We have demonstrated that the inability to secrete the water soluble glycoprotein form of the ABO blood group antigens into saliva is significantly more common in patients with Graves' disease than control subjects (40% vs 27%: P less than 0.025) but not among those with Hashimoto's thyroiditis or spontaneous primary atrophic hypothyroidism. Non-secretion is associated with increased susceptibility to infection and to asymptomatic carriage of some microorganisms. Although Yersinia enterocolitica has been found to express antigen cross reactive with the TSH receptor, we did not find an increased prevalence of Yersinia species in the faeces of 107 patients with Graves' disease. The isolation rate (less than 1%) was similar to that observed in the local population with diarrhoeal illness. Salivary IgA levels determined by whole cell ELISA with Y. enterocolitica 03 were not elevated in the majority of specimens examined. The results suggest that in contrast to reports from Scandinavia, there is no strong evidence that yersiniae play a role in the pathogenesis of Graves' disease among patients in South east Scotland. Non-secretors are significantly over represented among patients with several other autoimmune diseases; however, with the exception of antitubulin antibodies, non-secretors with Graves' disease did not have more antibodies to other human antigens than secretor patients.