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E. SCHIFFERDECKER, B. O. BOEHM and P. KÜHNL

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B. O. BOEHM, P. KUEHNL and K. SCHÖFFLING

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B. O. BOEHM, P. KUEHNL and K. SCHÖFFLING

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B. O. BOEHM, C. ROSAK and K. SCHÖFFLING

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E. SCHIFFERDECKER, B. O. BOEHM, P. KUEHNL and K. SCHÖFFLING

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B. O. Boehm, E. Schifferdecker, P. Kuehnl and K. Schöffling

Abstract. HLA-DR3 positive patients with Graves' disease (6 homozygotes, 7 heterozygotes, i.e. yielding 19 haplotypes) were studied by restriction fragment length polymorphism analysis using TaqI as restriction enzyme in order to look for polymorphisms in the HLA-DR3 allele of the human major histocompatibility complex. Polymorphic TaqI fragments of 11.6, 9.8 and 5.8 kb, each corresponding to HLA-DRβ sequences, were shown to differ in their prevalence in patients with Graves' disease and controls. The prevalence of DR3 polymorphisms in a total of 35 HLA-DR3-containing haplotypes was markedly different within patients with Graves' disease and Caucasian controls. Whereas a 11.6 kb fragment was rare in Graves' disease (2/19 haplotypes vs 8/16 in controls), the inverse ratio was found for a 9.8 kb fragment, with a prevalence of 17/19 and 8/16 haplotypes, respectively. A polymorphic fragment of 5.8 kb was exclusively seen in two DR3-containing haplotypes of patients with Graves' disease. Our data provide evidence that a DNA polymorphism of the HLA-DRβ genes, which is also reflected at the product level, is linked to Graves' disease.

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E. Schifferdecker, U. Ketzler-Sasse, B. O. Boehm, H. B. Ronsheimer, W. A. Scherbaum and K. Schöffling

Abstract.

Sera from 41 patients with Graves' ophthalmopathy were investigated for presence of autoantibodies directed against eye muscle preparations using different methods: 1. ELISA with pork eye muscle membrane preparations; 2. Immunoblotting with glycoprotein preparations from human eye muscle; 3. Indirect immunofluorescence with human eye muscle sections. The ELISA was not suitable for detection of specific immunoglobulin binding with sera from patients suffering from endocrine ophthalmopathy. Immunoblotting exhibited only nonspecific binding to some muscle proteins; it could be prevented by pre-adsorption procedures and was not different from the pattern observed with skeletal muscle as antigen. The indirect immunofluorescence technique revealed no binding of Graves' sera to human muscle sections, whereas sera containing antibodies against skeletal muscle bound to eye muscle as well. Thus far, an antigenic structure of eye muscle specific for Graves' ophthalmopathy is not detectable with the methodology used here. The possibility that retroorbital connective tissue may be the main target of the autoimmune process must be considered.

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W L Awa, B O Boehm, T Kapellen, B Rami, P Rupprath, W Marg, M Becker and R W Holl

Objective

To investigate HLA-DR genotype in association with chronological age or calendar year of disease onset and the time trend of genotype frequencies from 1969 to 2009. Additionally, to examine genotype frequency in relation to B-cell-, islet cell antibodies (ICA)-, autoantibodies to insulin-, insulinoma antigen 2 (IA2)-, glutamic acid decarboxylase-antibody positivity, thyroid antibody positivity, thyroid diseases or coeliac antibody positivity. Genotype associations with gender and ethnicity are also analyzed.

Subjects and methods

HLA-typed children and juveniles (n=1445) aged ≤20 years at disease onset from the German/Austrian DPV-database were examined. χ 2 statistics and mixed hierarchical logistic regression models were used to compare genotype frequencies and establish associations with age at disease manifestation, autoimmune antibodies/diseases, ethnicity and time trend.

Results

Subjects aged <6 years predominantly carried the genotype HLA-DR3/4 (25.2%), whereas in subjects aged >12 years the most prevalent HLA-DR genotype was X/X (18.1%). IA2 positivity was associated with HLA-DR4/X and HLA-DR3/4 positivity (P=0.011), and hypothyroidism was linked to HLA-DR4/4 (P=0.0103). More females carried the HLA-DR4/4 genotype (18.2 vs 12.7% P=0.0048) or were thyroid antibody positive (24.5 vs 14.7% P=0.0001). Larger numbers of <6 year olds were coeliac antibody positive than >12 year olds (14.8 vs 9.1% P=0.0037). No associations between migration background and B-cell-, thyroid- or coeliac-antibody positivity, and no time trend were found.

Conclusion

HLA-DR genotype associated with age at disease onset, ICA positivity and hypothyroidism; female gender with thyroid antibody positivity and low age of diabetes onset with coeliac antibody positivity.

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B.O. BOEHM, C. ROSAK, R. KUEHNL, E. SCHIFFERDECKER, E. JUNGMANN, K. SCHMIDT, P.-M SCHUMM-DRAEGER and K. SCHÖFFLING