Workshop No. 6: Histocompatibility - Disease

XI. Tagung der Gesellschaft fur Immunologie . 229

Workshop Nr. 6: Histocompatibility Disease

Medizinische Hochschule Hannover, Hannover, Federal Republic of Germany

6.1 The genetic basis of rheumatoid arthritis: population and family studies D. BRACKERTZ, W. MOLLER and P. WERNET

53 patients with rheumatoid arthritis (R. A.) were HLA-typed for Dw4 and DRw4. HLADw4 and HLA-DRw4 were significantly more frequent in patients (43% and 61% respectively) than in controls (17% and 19% respectively). In addition HLA-haplotypes were characterized in 71 members of 6 families in which at least one member had R. A .. The HLAantigens Dw4, DRw4 and Cw3 occurred with an increased frequency of 48% (34 of 71) and 31 % (22 of 71) respectively. 8 (24%) out of the 34 Dw4- and DRw4-positive family members were suffering from R. A .. Thus the relative risk for R. A. in relation to the presence of DRw4 in unrelated individuals and families was 6.5 and 11.4 respectively. In each multiple case family R. A. was associated with a specific familial HLA-haplotype, although not all members with this particular haplotype were affected with the disease. The importance of genetic factors for the pathogenesis of R. A. is further demonstrated by the fact that homozygozity for Dw4 and DR w4 is associated with a rapidly progressive and extraordinarily severe course of the disease.

I. Medizinische Klinik, Hautklinik und Hygiene-Institut, Universitat Kiel, Kiel, Federal Republic of Germany

6.2 HLA-related control of T-cell responses in vitro in psoriasis W. L. GROSS, I. VORWERK, E. CHRISTOPHERS, E. WESTPHAL and M. SCHLAAK

Blood lymphocyte preparations from 28 patients with psoriasis were tested for their ability to respond to mitogens and A-streptococcal antigens. The capacity to induce lymphocyte proliferation or lymphokine synthesis was tested by 3H-thymidine uptake or was detected by LIF activity with the indirect Clausen technique. The lymphocyte responses to PHA, ConA, and PWM were impaired (p
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XI. Tagung der Gesellschaft fur Immunologie

Medizinische Univ.-Poliklinik Munster, Federal Republic of Germany

6.3 Essential hypertension, a heritable disease linked to HLA-B17 H. W. INTORP, F. WESSELS and H. LOSSE

In previous studies it has been demonstrated that severe essential hypertension is a heritable disease in man as well as experimental animals. To determine whether it is associated with a HLA-antigen of the A, B or C locus 48 unrelated patients with the characteristic clinical symptoms of severe essential hypertension were tested for their HLA phenotype. In some instances family studies were performed using afflicted persons as well as family members without hypertension. The HLA-A, HLA-B and HLA-C antigens were determined by the lymphocyte microcytotoxity method using 68 standardised antisera to assess 32 specificities. 21 patients with severe essential hypertension had the HLA-BI7 antigen, as compared to 5 matched control subjects. In the family studies hypertensive patients had an even higher correlation with B17 than the group of randomly selected patients with essential hypertension. From this observation it can be concluded that essential hyper hypertension is a heritable disease in many instances linked to the HLA-BI7 on the chromosome 6. Testing for the presence of this antigen may help to secure the clinical diagnosis of this disease.

Immuno!. Lab., Mediz. Univ.-Klinik, Abt. II, Tiibingen, Federal Republic of Germany, and Basel Institute for Immunology, Basel, Switzerland

6.4 Immunogenetic aspects of natural human immunity against infections with group A streptococci D. LAUDIEN, D. BRAUN, W. RIESEN, F. SCHUNTER and P. WERNET

Natural aquired immunity in man against group A streptococci is accompanied by restriction against group A polysaccharide specific antibodies concerning the isotypic markers and the number of clonotypes visualized by isoelectric focusing. Persistence of clonal IgG-patterns over up to 11 years was shown and clearly demonstrates the existence of stable qualitative B-cell-memory within this system. Studies of families in which at least one member suffers from rheumatic fever or rheumatic heart disease revealed more complex segregation of the clonal antibodies than would be expected from simple mendel ian laws. The stimulation of peripheral blood cells by streptococcal A vaccine as well as the level and the heterogeneity of the clonal immune response were analysed for any association to HLAA, B, C, DR antigens and the genetic markers Pgm 3 , Bf, Glo in the case of healthy persons compared to patients suffering from rheumatic fever and rheumatic heart disease. An attempt is made to define a common genetic denominator for the immune response patterns to group A streptococci in man.

XI. Tagung der Gesellschaft fiir Immunologie . 231 Department of Medicine II and Institute of Blood Group Serology, University of Vienna, Austria, and Endocrine Department, Klinikum Steglitz, University Berlin, Federal Republic of Germany

6.S TSH-receptor and organ-specific autoantibodies in HLA-DR-typed insulin-dependent diabetics H. LUDWIG, G. SCHERNTHANER, H. SCHLEUSENER, B. WENZEL, P. KOTULLA and W. R. MAYR

Up to now no data are available regarding the incidence of TSH-receptor autoantibodies in insulin dependent diabetics. These antibodies might play an important role in the pathogenesis of Graves' disease and might occur either in patients with generalized autoimmune diathesis or independently of the other family of organ specific autoantibodies. Thus, the following questions should be elucidated in insulin-dependent diabetics: 1. incidence of TSH-receptor antibodies, 2. incidence of organ specific autoantibodies (adrenal, thyroid and gastric parietalcell antibodies) and 3. correlation between autoantibody production and IDD-associated HLA-antigens. 60 patients with insulin-dependent diabetes and 50 healthy individuals were studied. TSH-receptor antibodies were determined by a radio ligand receptor assay. Adrenal, thyroid and gastric parietal-cell autoantibodies by standard immunofluorescence technique, and HLA A, B, C and DR-antigens by standard NIH-microlymphocytotoxicity techniques. In 6 of the 60 patients unequivocal positive TSH-R-ab and in 7 TSH-R-ab with borderline titers were found, but in none 01 the controls. Thyroid microsomal antibodies were detected in lS%, thyroglobulin antibodies in 12%, gastric parietal cell antibodies in 15% and adrenal antibodies in 2 patients. By contrast, the respective incidence of these antibodies in controls was 6%, 7%, 3% and o. Adrenal and thyroid microsomal antibodies were predominantly found in HLA DRw3 and/or BS positive patients, whereas for the other antibodies tested no such tendency was found.

Immuno!. Lab., Dept. Med. II, University of Tiibingen, Federal Republic of Germany

6.6 HLA-DR serology in transplantation G. MOLLER and P. WERNET

Precise assessment of the HLA-D region as equivalent to the murine I region complex by serology has become an important component in donor selection for kidney and bone marrow transplantation. Five to ten highly specific anti-DR sera were employed for each defined HLA-DR alloantigen in complement mediated cytotoxicity, indirected immunofluorescence or a combination of both methods for double-labelling. Increasing serological sophistication has allowed finer judgement of D region identity or the definition of differences, such as splits in the DRW2 and DRW3 alleles as well as the uncovering of more than one DR locus. 32 kidneyrecipients typed for DR antigens within the South German Study Group could be followed for more than 6 months thus far. Graft survival was clearly better in cases of one or even two DR antigen matches. Within 3 months 50% of the patients who shared no HLA-DR antigen with their donor lost their graft. With the presently active blood transfusion protocol for all transplant candidates, of particular relevance seems to be the assessment of presensitization against HLA-ABC versus

232 . XI. Tagung der Gesellschaft fur Immunologie -DR antigens. By careful platelet and cell line absorption of alloantibody containing sera and subsequent testing on genotyped and isolated heterozygous and homozygous Band T cells it became clear that certain sera containing ABC-antibodies mimic the serological reaction pattern of anti-DR antibodies. This can lead to overlooking of presensitization and thus result in a wrong classification of a transplant patient with regard to transplant priority. Thus the method of 125I-IgG aggregate uptake by Fe-receptors and its inhibition by anti-DR antibodies was successfully employed for the serological dissection of ABC versus DR type antibodies. This analysis may force a reassessment of transplant priority lists with regard to the true sensitization of a transplant candidate.

2nd Department of Internal Medicine and Institute of Blood Group Serology, University of Vienna, Austria

6.7 Systemic lupus erythematosus (SLE) and HLA-DRw3 O. SCHERAK,]. S. SMOLEN and W. R. MAYR Immune response genes by analogy with the mouse system might be associated with gene products of the human HLA-D locus. Human B-cell alloantigens (HLA-DR) can be detected serologically and are closely related to the HLA-D antigens determined by mixed lymphocyte culture. HLA-DR antigens were examined in 28 patients with SLE and in 125 controls by the two-colour fluorescence method. HLA-DRw3 was significantly (p corr = 0,03) increased in SLE patients (39,2%) compared with controls (16%). The relative risk of SLE for HLA-DRw3 carriers is 3,39. Controls showed a higher frequency of HLA-DRw4 (23,2%) and HLADRw5 (26,4%) in contrast to SLE patients (DRw4: 3,5% and DRw5: 7,1%), but these differences were not significant. DRw3 positive SLE patients had a significant (p <0,05) later onset of their disease (43,9 ± 16,1 years) than DRw3 negative patients (32,9 ± 9,4 years). Analysis of the DRw data with regard to LE skin rash, Raynaud phenomenon, arthritis/ arthralgias, nephritis, serositis, psychosis/neurological manifestations, hemotological changes, lung involvement and vasculitis showed no significant associations. Antinuclear antibodies and n-DNA antibody binding capacity were monitored for more than 3 years (40,25 ± 20,16 months). The mean values of n-DNA antibody binding capacity were found to be not significantly different in the DRw3 positive and negative group. In contrast DRw3 negative SLE had a significant (p <0,00025) higher frequency of high titers of antinuclear antibodies (:~1 :320). The data indicate that immunogenetic factors are relevant to the development of SLE.

2nd Department of Internal Medicine, Institute of Immunology and Institute of Blood Group Serology, University of Vienna, Austria

6.8 B-cell alloantigens in rheumatoid arthritis (RA)

J.

S. SMOLEN, O. SCHERAK, E. ]. MENZEL and W. R. MAYR

Immunogenetic factors are discussed in the etiology of RA. Human B-Iymphocyte alloantigens which have homologies with the immune response associated antigens in murine systems and which are coded for by the HLA-DR locus were detected serologically with a panel of sera from the 7th International Histocompatibility Workshop in 70 patients with classical or definite RA and 125 controls. We observed a significant (p corr < 0,001, relative risk of 3,70)

XI. Tagung der Gesellschaft fUr Immunologie . 233 increase of HLA-DRw4 in patients with RA: DRw4 was present in 53% of RA and in 23% of the controls. We did not observe a correlation between onset of disease and the presence of HLA-DRw4. There was no association between HLA-DR antigens and serological (detection and titer of IgM-rheumatoid factor or antinuclear antibodies) or radiological (staging according to Steinbrocker) parameters. Another indicator of disease severity, namely necessity for systemic treatment with corticosteroids, showed also no relation to any HLA-DR antigen. In 22 of these patients, lymphocyte transformation to denatured type I collagen was performed and a significant transformation was observed in 15 patients. There was no significant correlation with any of the HLA-DR antigens. Likewise, no association was seen between HLA-DR antigens and the occurrence of antibodies to collagen. The association of RA with HLA-DRw4 suggests that immune response gene(s) may be involved in the development of RA but may be not relevant for severity of the disease.

Abt. Immunol. und Abt. Kinderheilkunde der Universitat Kiel, Federal Republic of Germany

6.9 Microabsorption studies of non-crossreactive extra reactions of HLA antisera with childhood leukemic cells E. WESTPHAL, U. LASSON and W. MOLLER-RuCHHOLTZ

Extra reactions of well-defined cytotoxic HLA antisera with childhood leukemic cells of ALL, AMMoL or AML origin may show cross reactive or non-crossreactive patterns. Antisera of the latter type, being reactive with >75% of the patient's cells, were absorbed respectively with leukemic cells or normal peripheral lymphocytes exhibiting corresponding HLA antigens, using our well established multiple micro absorption procedure. Findings: (1) Appearance of extra reactions only during the phase of acute disease, not in remission. (2) Titers of antisera with leukemic cells were regularily 3 - 4 steps above the titers with normal cells. (3) The reactions were independent of incubation temperatures. (4) They were complementdependent. (5) Under the following conditions activity could not be removed by 4 x 105 absorbing cells per 2 ~I: (a) absorption with normal cells/testing with leukemic cells; (b) absorption with leukemic cells/testing with normal cells; (c) absorption with leukemic cells/ testing with leukemic cells (confirmation by chessboard titrations with much larger amounts of cells). These data may be considered in relation to the findings of Bias et al. (Science 178. (1972), 304), though different in that we found new reactions which are cytotoxicity-positive and absorption-negative (CYPAN).