Antibodies to Yersinia enterocolitica in immunogenic thyroid diseases

In 1976 Shenkman et al. revealed that in patients with thyroid disorders antibodies against Yersinia enterocolitica could be demonstrated in increased frequency. In 1983 Ingbar et al. first established that the gram-negative bacterium Yersinia enterocolitica shows on its surface saturable binding sites for thyrotropin (TSH). If such binding sites resemble immunologically human TSH receptors this would indicate that TSH receptor antibodies could be produced in selected individuals having been infected with bacteria showing TSH receptors. The aim of our study was to compare the incidence of antibodies against Yersinia enterocolitica in two groups of thyroid disorders which are either immunogenic (Graves' disease and Hashimoto thyroiditis) or non-immunogenic (toxic adenomas, endemic goitre). In our series of 111 patients antibodies against Yersinia enterocolitica were demonstrated in a significantly higher percentage (36.3%) in patients suffering from immunogenic than in patients with non-immunogenic thyroid disorders (19.6%). The antibody titres were mainly directed towards Yersinia subtypes 8 and 3. It may, therefore, be assumed that the gram-negative bacterium Yersinia enterocolitica may have an active part in triggering immunogenic thyroid diseases such as Graves' disease or Hashimoto thyroiditis.     

Cell-mediated immunity in systemic lupus erythematosus

Cell-mediated immunity in systemic lupus erythematosus (SLE) was assessed by skin testing with six common antigens and by streptokinase-streptodornase (SK–SD) induced leucocyte migration inhibition in twenty-four SLE patients, who were age- and sex-matched with twenty-four healthy subjects or patients with diseases not known to be associated with immunological abnormalities. 25% of SLE patients were anergic, and the migration of their leucocytes was not inhibited in the presence of SK–SD. Depressed cell-mediated immune responses were significantly related to disease activity. Patients with inactive or mildly active SLE exhibited selective hyporeactivity to purified protein derivative (PPD), while those with moderately to severely active SLE had marked depression of cell-mediated immunity, as manifested by both skin testing with common antigens and leucocyte migration inhibition in response to stimulation by SK–SD. A significant positive correlation was found between absolute peripheral lymphocyte counts of the SLE patients and the number of their positive skin tests. Peripheral lymphocyte counts were significantly decreased in the anergic SLE patients and in those with moderate to severe disease activity. The correlation found between skin test reactivity and absolute lymphocyte count suggests lymphocytopenia as the mechanism of the immune suppression. On the other hand, lymphocytopenia alone cannot explain the depression of the leucocyte migration inhibition response to SK–SD observed in the SLE patients, because in this system a relatively constant number of lymphocytes is employed. In conclusion, the depression of cell-mediated immune responses in SLE is caused by both lymphocytopenia and lymphocyte hyporeactivity.     

Cellular and humoral autoimmune responses against human eye muscle membrane antigen in Graves' disease

75 patients with Graves' disease (54 with ophthalmopathy) were investigated using the tests of leucocyte adherence inhibition and immune adsorption with 125I-labelled Staphylococcus Protein A, against human eye muscle "crude" membrane antigen. The results of positive leucocyte adherence inhibition (10 out of 26 vs. 1 out of 28, P less than 0.05) and anti-human eye muscle membrane antibody index (mean +/- S.D.) (1.89 +/- 1.20 vs. 0.84 +/- 0.38, P less than 0.001) showed a correlation with the patients with clinically active eye disease and the HLA-B8 antigen in Graves' ophthalmopathy (P less than 0.01). Positive leucocyte adherence inhibition was observed in 9 out of 21 cases of Graves' disease without ophthalmopathy, but its prognostic relevance has to be confirmed in the development of ophthalmopathy.     

A study of lung-specific, cell-mediated immunity in chronic pulmonary diseases

Lung-specific, cellular hypersensitivity was studied in patients with chronic pulmonary diseases. To do this, a leucocyte migration inhibition assay was performed using patient and control leucocytes incubated in the presence and absence of a soluble lung extract. Control antigens consisted of liver and kidney extracts. The immunological reactivity of these control antigens was tested by measuring migration, subsequent to incubation of leucocytes from patients with hepatic or renal disease with the liver and kidney extracts, respectively. As a second in vitro test of cell-mediated immunity, a lymphocyte cytotoxicity assay was performed, using as target cells normal chromium-labelled human lung cells. Leucocyte migration inhibition was produced by the lung extract in four out of six patients with chronic bronchitis, five out of seven patients with tuberculosis, one out of five patients with sarcoidosis and in the single patient studied with rheumatoid lung disease. Inhibition did not occur with the leucocytes from control subjects. Neither was migration inhibited when leucocytes from the patients with lung disease were incubated with the control antigens. The reactivity of these control antigens was confirmed by the fact that leucocyte inhibition occurred in two out of three patients with hepatic disease, studied using the liver extract, and in two out of three patients with renal disease, studied with the kidney extract. The cytotoxic potential of anti-lung lymphocytes was demonstrated by the fact that significant target cell cytotoxicity occurred using lymphocytes from two out of four patients with chronic bronchitis, four out of five patients with tuberculosis and in two out of four patients with sarcoidosis.     

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.     

Comparative study on IgG and IgA antibodies against human thyroid and eye-muscle antigens in Graves' ophthalmopathy.

Circulating IgG and IgA anti-thyroid and anti-eye muscle antibodies were investigated in 87 patients with Graves' disease (60 cases with ophthalmopathy). The ELISA method was used. Both IgG and IgA antibodies were demonstrated against human thyroid and eye-muscle membrane or cytosol antigens. Anti-eye-muscle antibodies of the IgA type were observed more frequently than those of the IgG type (25 cases vs. 18 were demonstrated with membrane antigens and 37 cases vs. 23 with cytosol antigens). The respective distributions for thyroid antigens the cytosol fraction were 55 cases vs. 13 and 18 cases vs. 36. A significant difference was observed in the anti-thyroid IgG levels and the anti-eye-muscle membrane or cytosol levels between the patients with Graves' disease and those in control group (P less than 0.001). The difference in the IgA antibody to thyroid and eye-muscle antigens was significant between the patients with and without ophthalmopathy (P less than 0.002). The strong correlation between the levels of IgA antibodies to thyroid and those to the eye-muscle cytosol fractions might be connected with the theory of the common aetiology of the thyroid and eye diseases in Graves' ophthalmopathy (P less than 0.001). Circulating IgA anti-human thyroid and eye-muscle antibodies seemed to have a diagnostic relevance in the development of ophthalmopathy in Graves' ophthalmopathy.     

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。     

The effect of hydrocortisone on the incorporation of tritiated thymidine by human blood lymphocytes cultured with phytohaemagglutinin and pokeweed mitogen

Leucocyte migration inhibition tests (LMT) with rat and human liver mitochondria are reported in forty-nine thyroid patients and forty-seven healthy controls. Whereas normal subjects and colloid goitre cases were inactive in this test, patients with autoimmune thyroiditis and thyrotoxicosis gave positive mitochondrial reactions which paralleled the organ-specific thyroid microsomal LMT obtained in the same patients and were not species dependent. The active antigen may be in the inner membranes of the particles.

As with thyroid microsomes, intense inhibition with mitochondria was seen in the hypercellular variant of Hashimoto goitre characterized serologically by low or absent thyroglobulin antibodies, and the lowest LMT values occurred in the rare cases showing poor response to thyroxine therapy. An inverse correlation was found between mitochondrial LMT and thyroglobulin antibody titres. Surprisingly, weak LMT was also found in four thyroid patients who happened to have mitochondrial antibodies in the serum in addition to the usual thyroidspecific reactivities. The mitochondrial LMT appears to be of widespread occurrence in autoimmune diseases and also develops following tissue injury. Its possible significance in relation to cellular immunity, cell destruction and the inflammatory response is discussed.

     

Studies of cellular and humoral immunity in typhoid fever and TAB vaccinated subjects.

An assessment of both humoral and cell-mediated immune responses to Salmonella typhi antigens in patients with acute typhoid infection, TAB inoculated subjects and in healthy controls is reported. Cell-mediated immunity as assessed by the leucocyte migration inhibition test (LMI), and developed in all cases with typhoid fever. Positive LMI was evident in the first week of the illness and was maintained during the evolution of disease and in some patients was still present after a year. It also developed at the end of 3 weeks in five out of nine TAB vaccinated subjects. Weakly positive LMI was noticed in only two of twenty asian and caucasian controls. Antibodies, determined by the standard Widal test, were significantly raised in both patients with typhoid fever and TAB inoculated subjects. The antibodies and cellular reactivity developed almost simultaneously but there was no correlation between the agglutination titres and LMI positivity, implying that they are independent of each other. Typhoid patients also showed significantly raised serum IgM and IgA levels and increased concentrations of secretory IgA in their saliva.     

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.     

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.     

冷冻保存人甲状腺细胞的活力,功能和临床应用

本文研究冷冻保存不同时间甲状腺细胞的活力、功能及其在甲状腺刺激抗体（ＴＳＡｂ）检测中的应用。结果表明：冻存３、６和１２个月的正常或甲亢甲状腺细胞具有良好的活力，各冷冻时相的细胞对ＴＳＨ刺激生成ｃＡＭＰ的反应性无明显差异，且与未冷冻细胞的反应性相似。以冻存甲亢甲状腺细胞测定血清ＴＳＡｂ．在Ｇｒａｖｅｓ病（ＧＤ）未治组的检出率为０．９１３，而甲状腺腺瘤（甲瘤）组与对照组ＴＳＡｂ均为阴性。这提示冻存一年内的人甲状腺细胞仍保持较好的活性与功能，可以作为甲状腺功能和甲状腺疾病基础与临床研究的材料。     

Yersinia enterocolitica and thyroid autoimmunity

The concordance rate of Graves' disease in pairs of identical twins of 30 to 60% points to the influence of environmental factors, and infections have often been incriminated in the pathogenesis. More than 15 years ago we demonstrated an increased frequency of antibodies to Yersinia enterocolitica (Y. ent.) serotype 3, which later was confirmed by others, suggesting a link between infection with Y. ent. and autoimmune thyroid diseases.     

Thyrogastric autoimmune disease. Studies on the cell-mediated immune system and histocompatibility antigens.

Cell-mediated immune responses were studied in autoimmune diseases of thyrogastric type, Hashimoto's thyroiditis and autoimmune pernicious anaemia-type gastritis. Specific cell-mediated immunity was investigated by the leucocyte migration inhibition procedure, and general cell-mediated immunity (T-cell performance) was studied by standard in vivo and in vitro tests. In thyrogastric autoimmune diseases inhibition of migration of leucocytes was induced by thyroglobulin and gastric parietal cell microsomes; under conditions of presumably low cellular sensitization, stimulation of migration was observed. There was no depression of general cell-mediated immunity, in contrast to what occurs in systemic lupus erythematosus and related autoimmune diseases. A weak association of autoimmune gastritis with HL-A3 and HL-A7 (P LESS THAN 0.05) lost significance when an appropriate correction was applied; this weakness with HL-A clearly does not explain the strong genetic component in thyroid and gastric autoimmunity.     

Cell-mediated immunity (CMI) to human wart virus and wart-associated tissue antigens.

Lymphocyte transformation (LT) and leucocyte migration inhibition in agarose were used to demonstrate cell-mediated immune response to purified human wart virus and wart tissue extract in various subjects with warts and those without past history of warts. Most individuals bearing warts for less than 1 year duration showed positive cell-mediated responses to both the virus and tissue extract whereas very few of those who had warts for longer duration responded to either antigenic preparation. The difference was statistically significant. Subjects who had warts in the past also showed positive responses but these tended to decrease in degree with time. Surprisingly a group of subjects who never had warts before also responded to stimulation with the virus, but not to the extract. The positive response to stimulation with wart tissue extract reflects the presence of wart associated antigens other than the virus. Cell-mediated immunity against the wart virus and wart-associated antigens is probably important in preventing the persistence or even establishment of disease but this protective immunity is short-lived. The lack of quantitative correlation between LT and leucocyte migration inhibition demonstrable in this study suggests that these two are separate events in in vitro lymphocyte stimulation with antigens.     

Long-term pathologic changes of alpha1-acid glycoprotein (orosomucoid) glycoforms in autoimmune thyroid disease

OBJECTIVE: We measured alpha1-acid-glycoprotein (AGP) in patients with autoimmune thyroid disease to study a possible relationship between microheterogeneity of the naturally occurring glycoforms of AGP and autoimmune thyroid disease. DESIGN, PATIENTS, MEASUREMENTS: In a group of 12 fasting thyrotoxic patients (11 females, mean age: 43 years) with newly diagnosed Graves' disease (subgroup 1), we measured serum concentrations of total AGP and its 3 glycoforms (micromol/l, crossed affinity immunoelectrophoresis with con A in the first dimension gel) as well as total thyroxine, total triiodothyronine, thyrotropine, thyroid peroxidase antibodies (anti-TPO), antibodies against the TSH receptor (TRAb, TRAK), at baseline and after 12 months of antithyroid drug therapy (ATD). For comparison, 4 subgroups of thyroid patients (patients with Graves' disease and thyroid associated ophthalmopathy (TAO) (subgroup 2, n = 10), radioiodine treated Graves' patients (subgroup 3, n = 7), Graves' patients without TAO (subgroup 4, n = 13), patients with Hashimoto's thyroiditis (subgroup 5, n = 8)) and 25 normal controls (17 females, mean age: 38 years) were studied. RESULTS: In subgroups of TRAb positive Graves patients' serum levels of glycoform 1, 2 or 3 increased significantly (p < 0.005) after 12 months of ATD as compared to both baseline of that person or normal controls. No significant changes were found in the TRAb negative Hashimoto subgroup. CONCLUSION: Patients with autoimmune Graves' disease changed their relationship to AGP, and thus a role of AGP and its 3 glycoforms is suggested in the pathogenesis of Graves' disease.     

Isolation of Mallory bodies and an attempt to demonstrate cell mediated immunity to Mallory body isolate in patients with alcoholic liver disease

Mallory bodies were isolated from necropsy livers from patients with alcoholic hepatitis with and without cirrhosis with a Ficoll viscosity barrier. The purity of Mallory bodies in the isolate varied between 70 and 90%, estimated by counting Mallory bodies and non-Mallory body structures in haematoxylin-eosin stained smears. Electron microscopy confirmed the presence of Mallory bodies in the isolates. The Mallory body isolate was used as antigen in the agarose leucocyte migration inhibition test in order to test the cell-mediated immunity. No significant difference in leucocyte migration was found between controls and patients with alcoholic hepatitis, alcoholic steatosis, alcoholic cirrhosis and miscellaneous liver diseases.     

Accumulation of thyroid antigen-reactive T lymphocytes in the gland of patients with subacute thyroiditis.

Blood and thyroid-infiltrating lymphocytes from patients with de Quervain's subacute thyroiditis were tested in the leucocyte migration inhibition test for cell-mediated immunity (CMI) to thyroid antigen. Blood leucocytes were positive for CMI in eleven out of thirteen (85%) patients in the acute phase of the disease. In five cases tested repeatedly this reactivity disappeared after 7-11 months. Thyroid-infiltrating lymphocytes were obtained from eight patients by fine-needle aspiration biopsy. There was a relative accumulation of T lymphocytes (90 +/- 5%) in the thyroid gland as compared to lower numbers of T cells (65 +/- 5%) in the blood. When thyroid-infiltrating lymphocytes were tested for CMI to thyroid antigen, a significantly stronger inhibition was demonstrated with the infiltrating lymphocytes as compared to that of blood leucocytes. We conclude that thyroid antigen-reactive T cells accumulate in the glands of patients with subacute thyroiditis. This observation is in accordance with the presumed viral aetiology of this disorder.     

Evaluation of immunological tests in thyroid diseases

We evaluated immunological tests for autoimmune thyroid diseases. Although both humoral and cellular immunity are correlated to the onset and pathophysiological progression of these diseases, the major tests employed in daily clinic belong to the former. We measured the anti-TSH receptor antibody (TRAb, TBII), circulating immune complex (CIC), thyroid growth stimulating immunoglobulin (TGSI) and interleukin-2 (IL2) levels in patients with Graves' disease (GD) and chronic thyroiditis (CT). The normal range of TRAb in 190 healthy controls was from -10.9 to +10.3% calculated from the mean +/- 2SD. Sixty eight out of 78 untreated cases of GD (87.2%) showed a higher TRAb than the upper normal level (positive), 92 out of 131 cases of GD under treatment (70.2%) were positive and only 5 out of 57 cases of treated GD (8.8%) were positive. Three neonates out of 12 GD mothers had a positive TRAb and one of them developed neonatal GD. Nine out of the 45 CT (20%) had positive TRAb, but about half were euthyroid and goitrous. In untreated GD, CIC was distributed widely from the normal range to high levels. CIC showed a significantly negative correlation with TRAb. TGSI correlated with goiter size and CIC in GD revealed autologous inhibition on TGSI. Three cases showed markedly decreased levels of TRAb which was found to be due to anti-TSH antibodies. Production of IL2 in GD was impaired, but it was improved by treatment of GD.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increase in peripheral natural killer cell activity in patients with autoimmune thyroid disease.

Changes in the activity and number of natural killer (NK) cells in peripheral blood in patients with autoimmune thyroid disease were examined. NK activity was measured in a 4-hr 51Cr-release assay and the number of NK cells was analyzed with FITC-conjugated monoclonal antibodies by use of an automated flow cytometer. NK activity in patients with untreated Graves' disease (n = 25, 39.7 +/- 13.5%, P less than 0.05) and Hashimoto's thyroiditis (n = 18, 41.0 +/- 14.2%, P less than 0.05) was high compared to the activity in non-pregnant controls (n = 61, 32.6 +/- 15.0%). NK activity in patients with postpartum Graves' thyrotoxicosis (n = 11, 48.6 +/- 18.9%) was markedly increased compared to the activity in non-pregnant controls (P less than 0.01) and in postpartum controls (n = 29, 33.8 +/- 15.2%, P less than 0.05), although the mean ages of each group did not differ significantly. Moreover, NK activities in the thyrotoxic state were significantly higher than those in the euthyroid state in the same patients with postpartum Graves' thyrotoxicosis or with postpartum destructive thyrotoxicosis. The number of CD16 positive cells increased in patients with postpartum Graves' thyrotoxicosis. However the number of CD16 and CD57 positive cells were normal in all other groups of patients. These results indicate that an increase of NK activity is associated with exacerbation of autoimmune thyroid disease both in Hashimoto's thyroiditis and in Graves' disease and suggest that NK cells might have an important role for the control of disease activity in autoimmune thyroid disease.