The genetics of Graves' disease: HLA and disease susceptibility.

To relate genetic variation in Graves' disease (GD) susceptibility to polymorphism at MHC loci, clinical and family studies were undertaken in eastern Hungary. Among 1980 relatives of 534 index patients, 2.9% of siblings, 2.7% of offspring, and 3.0% of parents had GD. HLA haplotype combinations in affected sibling pairs were determined in the present data and combined with data in the literature (12 sibling pairs from Farid 1981, 12 from Chan et al. 1980, and 15 from Sasazuki et al. 1983); 43, 23, and 1 affected sibling pairs shared, respectively, 2, 1, and 0 HLA haplotypes. This distribution is inconsistent with simple dominant inheritance, but is consistent with simple recessive inheritance of HLA-related susceptibility over a range of gene frequencies (0.2-0.4). A frequency of 0.3 gives the best fit and is consistent with penetrance of 7.1% for the recessive susceptibility genotype; the data, however, can accommodate penetrance values up to 16%. The distribution of HLA haplotypes in 33 families related disease susceptibility more strongly to DR than to other loci. The distribution of HLA-B8 genotypes in 256 patients was in close agreement with Hardy-Weinberg equilibrium proportions, also favoring recessive inheritance of MHC-related susceptibility. The probability that an individual will be affected with GD can be predicted, based on sex, HLA genotype, and family history. For example, 14.9% of DR3-positive women with an affected first degree relative are likely to be affected. These predictions can be tested as family data accumulate.

Cytotoxic endothelial cell antibodies in mixed connective tissue disease.

We examined the presence of anti-endothelial cell antibodies in the sera of 44 patients with mixed connective tissue disease (MCTD). Warm antibodies against endothelial cells were found in 45.4% of patients; the presence of these antibodies was positively correlated with anti-monocyte antibodies (P less than 0.01) but not with anti-lymphocytic antibodies. Strong correlations were found between the presence of these antibodies and abnormalities of pulmonary ventilatory capacity, neurophysiologic and myocardial function. Anti-endothelial antibodies were related to the high rate of spontaneous abortion noted in female MCTD patients. The identification of the antigen identified by MCTD sera in endothelial cells would help in understanding disease manifestations.

Methimazole blocks Graves' IgG but not interferon-gamma HLA-DR expression by thyroid cells.

We have expanded our early observation that a potent Graves' IgG preparation induces HLA-DR expression on thyroid cells, by showing that four randomly-selected Graves' IgG's were also capable of inducing DR antigens on thyroid cells. The effect of Graves' IgG was specific to thyroid cells, as it did not induce MHC Class II molecule expression on endothelial cells whereas interferon-gamma and immune complexes did so. The anti-thyroid drug methimazole was capable of rapidly reducing Graves' IgG-induced DR expression but to a much lesser extent than brought about by interferon-gamma. We conclude that Graves' IgG propagates thyroid-specific autoaggression by continued induction of DR antigens and than an important means whereby methimazole brings about remission is by reducing this induction.

Expression of HLA-DR antigens by thyroid cells: the effect of Graves' IgG.

We have investigated factors which influence HLA-DR expression on thyroid cells. While bTSH (100 mU/ml) did not enhance HLA-DR expression, it increased when brought about by IFN-gamma. Graves' IgG showed a dose-dependent (0.1-2 mg/ml) increase in DR expression and at a concentration of 2 mg/ml prolonged the time for which DR was expressed. The pathway of DR induction by Graves' IgG apparently differs from that by IFN-gamma. The humoral response in Graves' disease, by inducing DR expression, may be instrumental in propagating thyroid specific autoimmunity.

B lymphocyte subsets in Hashimoto's thyroiditis.

Two B lymphocyte subsets are identified on the basis of possession or lack of a surface molecule, CD5. The CD5+ B lymphocytes synthesize autoantibodies and in the process rearrange proximal variables of immunoglobulin genes. We have here studied the proportion and absolute counts of CD5+ B lymphocytes in the peripheral blood of 31 patients with Hashimoto's thyroiditis and related the findings to their HLA phenotypes and clinical features. Although the percentage and absolute number of surface immunoglobulin-positive (B) lymphocytes was comparable in patients to those in twenty controls, % CD5+ was significantly higher in the patient group (38.1 +/- 11.6 [+/- S.D.] vs. 27.9 +/- 10.1, p = 0.009). The absolute CD5+ cells (microliters) were also higher in patients with Hashimoto's thyroiditis (77.90 +/- 37.50 vs. 55.1 +/- 29.8, p = 0.020). The proportion of CD5+ cells was even higher in HLA-DR3 positive patients (43.1% +/- 7.2, n = 13) compared to six DR3+ controls (26.17% +/- 9.7, p = 0.0005). The difference between DR3- patients and controls was not significant (28.6% +/- 10.5 vs. 34.4% +/- 13.0). As the CD5 molecule may be induced on activated B lymphocytes, this study suggests that Hashimoto's thyroiditis is associated with an increase of activated B lymphocytes engaged in autoantibody synthesis. This defect is particularly obvious in DR3+ patients.

HLA-DR1 is associated with vitiligo.

In Eastern Hungary, vitiligo is found to be associated with HLA-DR1. When other autoimmune disorders are also present, DR3 is also increased.

Cytogenetic analyses of three papillary carcinomas and a follicular adenoma of the thyroid.

Cytogenetic data of three papillary carcinomas and a follicular adenoma using direct preparations or cell cultures or both after 7 to 60 days in vitro are presented. Although karyotype of the follicular adenoma proved completely normal, in each of the three papillary carcinomas a modal chromosome number in the diploid range and a deleted 11q were observed. In case 1 the del(11)(q23) was associated with rearrangement of chromosome 1 and other marker chromosomes. Our results suggest that 11q deletion may be specific for papillary carcinoma of the thyroid.

No mutations in the translated region of exon 1 in the TSH receptor in Graves' thyroid glands.

We have amplified a 285 base pair by nested polymerase chain reaction 38 nucleotide downstream from the most 5' transcription initiation site (7) encoding 55 of the 57 residues of exon 1 of the human TSH receptor. These DNA were amplified from 10 tissue blocks of thyroid tissue removed at subtotal thyroidectomy from 10 patients with Graves' disease. The amplified 285 nucleotide fragments were sequenced in search of mutations in the coding region of exon 1 and polymorphism in the 120 nucleotides of the untranslated region upstream of the first ATG codon. No such variations were found. We conclude that the polymorphism or mutation of the part of the TSH receptor extracellular domain encoded by exon 1 and of sequences immediately upstream of the first ATG codon are not relevant to the pathogenesis of Graves' disease.

Immunological investigations on patients with transplanted cornea: the role of tissue antigens in keratoplasty.

After briefly describing the conceptions of the HL-A antigens and of histocompatibility, the authors give an account of their own retrospective examinations in connection with keratoplasty. Transplantations between identical groups of the ABO-system have a greater probability of ending successfully, if at least 2 HL-A antigens of donor and recipient are identical; this being chiefly important when grafting is carried out into unfavourable, vascularized surroundings.

Examination of HL-A antigens and lymphocytotoxic antibodies in discoid lupus erythematosus.

The distribution of HL-A antigens and lymphocytotoxic antibodies has been studied in the sera of patients suffering from discoid lupus erythematosus (DLE). A significant deviation in HL-A5 antigen has been found in the 33 cases investigated, while the lymphocytotoxic antibodies were present in only 8 sera. The possible cause of these findings are discussed.

Tissue antigens and cytotoxic antibodies in polycythaemia rubra vera.

The distribution of tissue antigens for 22 antigens was studied in 46 patients suffering from polcythaemia rubra vera (PRV), using antibody containing sera from 80 multigravidae. The incidence of HL-A5 was significant higher and that of HL-A7 significant, while the frequency of HL-A13 was remarkably lower than in the normal population. No antibodies against erythrocytes, thrombocytes or lymphocytotoxic ones could be demonstrated.

Clinical relevance of cytotoxic antibodies occurring in pregnancy.

Cytotoxic antibodies were studied in 809 pregnant women's sera. Two kinds of antibody population: HLA-A, B, C and Ia-like, were detected and a correlation was established between their presence and some clinical conditions, such as preterm delivery, spontaneous abortion, pre-eclampsia, neonatal hyperbilirubinaemia and congenital anomalies. An opposite effect of HLA-A, B, C and Ia-like antibodies was observed as far as the first three parameters are concerned. On the basis of these investigations a protective role of Ia-like antibodies for the newborn can be implied.

Lymphocytotoxic antibodies in lightdermatoses.

Occurrence of lymphocytotoxic antibodies of the "cold type" was studied in 92 cases with various lightdermatoses. They were detected in the sera of 39% of the patients with cutaneous porphyrias and never in cases with polymorphous light eruption and other photodermatosis. Correlations between their presence and the duration of the porphyria as well as the severity of hepatopathy could be observed. The results indicate the importance of a long-standing tissue damage in the production of these antibodies. In addition the findings confirm the hypothesis according to which polymorphous light eruption does not belong to the lupus erythematosus entity.

Identification of lymphocyte subsets in peripheral blood and thyroid gland in Graves' disease by alpha-naphthyl-acetate-esterase reaction.

T lymphocyte subsets were prospectively examined in the peripheral blood and thyroid aspirates of 10 patients with hyperthyroid Graves' disease before and after treatment with methimazole and attainment of euthyroidism. T lymphocyte subsets were identified with monoclonal antibodies and pattern of alpha-naphthyl-acetate esterase (ANAE) staining pattern in the case of peripheral blood and ANAE staining pattern with thyroid aspirate smears. Before treatment, OKT8+ lymphocytes were significantly decreased (18.4% +/- 4.8) (S. D.) in the patients compared to control (28.8 +/- 6.7%, p less than 0.05), the OKT4/OKT8 ratio was increased (2.92 vs 2.11). Percent OKT8+ lymphocytes were not different from the controls when the ten patients had been rendered euthyroid. ANAE mononuclear cells with a diffuse pattern (presumed suppressor cells) were 4.2% +/- 1.8 before treatment and 8.3 +/- 2.4 (p less than 0.05) after treatment and 11.5% +/- 2.2 in controls. ANAE mononuclear cells with diffuse pattern represented 4.2% +/- 1.8 of the mononuclear cells infiltrating the thyroid gland of untreated patients and rose to 8.3% +/- 2.4 after the patients had become euthyroid. ANAE negative cells (B cells and some T cells) were increased in the thyroid of untreated patients. It is concluded that mononuclear cells with presumed suppressor T cell phenotype are decreased in the blood and thyroid glands of patients with active Graves' disease and that this defect is corrected when euthyroidism has been established.

Interplay of immunoglobulin G heavy chain markers (Gm) and HLA in predisposing to systemic lupus nephritis.

We have studied the distribution of IgG heavy chain markers (Gm) among 90 Hungarian patients with systemic lupus erythematosus (SLE) (55 of whom were also typed for HLA). This study confirms previously described increases in HLA-B8 and DR3 in this condition. No difference in the distribution of Gm phenotypes was found between patients and 168 controls from the same geographical area. HLA-B8/Gm homozygous individuals were, however, at greater risk for SLE (relative risk = 5.13) compared to B8 + Gm heterozygotes or B8- individuals, irrespective of Gm phenotype. When patients with renal manifestation (n = 40) were compared to those without, the Gm phenotype 3; 5, 13 was found to be significantly increased (chi 2 = 10.36, P less than 0.0001, relative risk (RR) = 4.69). HLA and Gm increased additively the risk for renal manifestations in that for those patients who were both Gm3;5,13+ and HLA-B8+, PR was 110, while it was 21.2 for Gm3;5, 13-/B8+, 7.9 for Gm3;5, 13+/B8- and 1.0 for Gm3;5, 13-/B8- patients. The study suggests that combined HLA and Gm typing can be used to identify SLE patients at high risk for manifesting renal abnormalities.

HLA-DR associations with Graves' disease in eastern Hungary.

We have typed 196 patients with Graves' disease from eastern Hungary (an iodine-deficient region) for HLA-A, -B, and -C antigens and 80 of the same patients for HLA-DR antigens. Our data confirm the previously described association of HLA-B8 and DR3 with Graves' disease, particularly in patients with ophthalmopathy. We also describe several new findings: an increase in the prevalence of HLA-BW35, particularly in combination with B8, among patients with ophthalmopathy and a slight increase in the prevalence of B8, DR7 among patients with ophthalmopathy. These results support the concept that MHC-linked susceptibility to Graves' disease is dose-dependent. Our results do not confirm a report suggesting that the prevalence of HLA-DR5 is increased among patients without ophthalmopathy in iodine-deficient areas: we found that the prevalence of HLA-DR5 was slightly decreased among patients with Graves' disease, with and without ophthalmopathy, in comparison with controls.