In this report, a new HLA-B locus antigen is described (tentatively called POT). The antigen has been identified with antisera against the antigens that belong to the B7-cross reacting group. In a collaborative study, based on exchange of cells and sera, it was confirmed by population and family studies that the antigen is distinct from B7, Bw42 or Bw73 and is associated with Bw6. In absorption experiments with EBV-transformed cell lines, the POT-antigens removes preferentially anti-B7, Bw42 and Bw73 activity and to a lesser extent antibodies reactive with the B40 CREG-antigens.

In the past 15 years, 411 sporadic narcolepsy patients have been diagnosed in the Hephata Klinik, Schwalmstadt, Germany. They were explored for presence or absence of excessive daytime sleepiness and narcolepsy in their relatives. A subset of 39 patients were explored for presence or absence of parasomnias. Six patients had more than one relative affected by narcolepsy-cataplexy. Forty-seven family members were investigated with the Stanford Center for Narcolepsy Sleep Inventory and a standardized parasomnia questionnaire. Twenty-four relatives had nocturnal polysomnographies and Multiple Sleep Latency Tests. HLA class I typing was performed in all sporadic and familial cases, class II and microsatellite typing was performed in all members of multicase families. Based on the Finnish prevalence study by Hublin et al., 1994, the relative risk for first degree relatives to develop narcolepsy-cataplexy was in our sample 16.5, 34.2 for excessive daytime sleepiness and 426.9 for parasomnias. Cataplexy, excessive daytime sleepiness and single narcoleptic symptoms in the multicase families segregate with the DRBI*1501, CARII:200, CARI: 103, DQBI*0602 haplotype. In two families, members with narcolepsy and isolated symptoms have inherited the DRBI*1501/DQBI*0602 haplotype from the nonaffected parent. The observed segregations in these two families may support the view that narcoleptic symptoms are expressed by DRBI*1501/DQBI*0602 carriers, independent of haplotype origin. Parasomnias do not segregate with a specific haplotype. The frequency of parasomnias in narcolepsy is much higher than in the general population. The empirical risk for first degree family members of narcolepsy patients to develop cataplexy seems to be low, whereas it is higher for EDS and highest for parasomnias.

While HLA class II alleles identification by means of complement mediated lymphocytotoxicity (serology) is almost replaced by DNA typing techniques, serology is still widely used for routine class I typing. The aim of this prospective study was to compare PCR-based Amplification Refractory Mutation System with serology in clinical HLA class I alleles assignment in patients receiving marrow transplants and their potential donors. The total discrepancy rate in 114 consecutively typed individuals for HLA-A and HLA-C alleles was only in favor of ARMS-PCR, whereas HLA-B typing was discrepant also in favor of serology. The discrepancies were higher in patients, particularily in those with acute lymphoblastic leukaemia, than in healthy individuals. We conclude, that ARMS-PCR is clearly superior to serology in definition of class I alleles, which might be of clinical importance particularly for bone marrow transplantation.

Narcolepsy is a sleep disorder that has been shown to be tightly associated with HLA DR15 (DR2). In this study, 58 non-DR15 patients with narcolepsy-cataplexy were typed at the HLA DRB1, DQA1 and DQB1 loci. Subjects included both sporadic cases and narcoleptic probands from multiplex families. Additional markers studied in the class II region were the promoters of the DQA1 and DQB1 genes, two CA repeat polymorphisms (DQCAR and DQCARII) located between the DQA1 and DQB 1 genes, three CA repeat markers (G51152, T16CAR and G411624R) located between DQB 1 and DQB3 and polymorphisms at the DQB2 locus. Twenty-one (36%) of these 58 non-DR15 narcoleptic patients were DQA1*0102 and DQB1*0602, a DQ1 subtype normally associated with DRB1*15 in DR2-positive narcoleptic subjects. Additional microsatellite and DQA1 promoter diversity was found in some of these non-DR15 but DQB1*0602-positive haplotypes but the known allele specific codons of DQA1*0102 and DQB1*0602 were maintained in all 21 cases. The 37 non-DQA1*0102/DQB1*0602 subjects did not share any particular HLA DR or DQ alleles. We conclude that HLA DQA1*0102 and DQB1*0602 are the most likely primary candidate susceptibility genes for narcolepsy in the HLA class II region.

Die Narkolepsie ist die Erkrankung mit der stärksten Assoziation mit dem HLA-System. Nur wenige Patienten haben bei der Typisierung der «Narkolepsie-typischen» HLA-Allele DRB1*1501 (DR15(2)) und/oder DQB1*0602 (DQ6(1)) einen negativen Befund. Bei Patienten und Gesunden in verschiedenen Populationen bestehen ethnische Unterschiede im Hinblick auf gefundene HLA-Haplotypen. Die Aussagekraft immungenetischer Daten für die Narkolepsie erfordert deshalb eine eindeutige klinische Charakterisierung der Patienten einschließlich Schlaflatenz-Tests und Verwendung standardisierter Fragebögen. Vergleiche der immungenetischen Daten von japanischen, kaukasischen und afro-amerikanischen Patienten legen nahe, daß die Assoziation der Erkrankung eher in der HLA-DQ (DQB1*0602) als in der DR Region (DRB1*1501) lokalisiert ist. In Familien mit Mehrfacherkrankungen (Multiplex-Familien) ließ sich kein einheitlicher Verberbungsmodus eines «Narkolepsie-Empfänglichkeitsgens” im HLA-Komplex festlegen. Patienten in Multiplex-Familien wurden in 21–35% DRB1*1501 negativ gefunden. Drei konkordante Zwillingspaare mit Narkolepsie waren ebenfalls DRB1*1501 negativ. Die Wahrscheinlichkeit für Verwandte 1. Grades von Narkolepsiepatienten, ebenfalls zu erkraken, ist 40mal, bei Familien mit mehreren Narkolepsiekranken bis zu 700mal höher als in der gesunden Bevölkerung. Die immungenetischen Daten lassen insgesamt darauf schließen, daß neben dem HLA-System andreren genetische bzw. nichtgenetische Faktoren für die Pathogenese der Narkolepsie eine Rolle spielen.

With the growing development and routine use of high-resolution DNA typing techniques for HLA class II alleles the importance of mixed lymphocyte cultures (MLC) with respect to bone marrow transplantation is under controversial discussion. Therefore, results of MLC assays performed for donor/recipient pairs were analysed in comparison to the degree of histocompatibility as determined by prospective serological and retrospective DNA typing. Three different groups were evaluated for MLC data: (A) siblings sharing two parental haplotypes, (B) related donors sharing one haplotype, with 1-3 MM on the second haplotype, (C) phenotypically identical unrelated donors. In group A, all MLCs were negative. In contrast, 55 out of 59 MLCs in group B and 30 out of 41 in group C were positive. In group C, 11 MLCs were positive despite phenotypic class I and DRB 1 compatibility. We conclude from our results that, at present, MLC is not mandatory for genotypically HLA-identical siblings while for donor/recipient pairs of groups B and C MLC should be carried out even in cases of phenotypical compatibility for class I and DRB 1 alleles.

PROBLEM:The role of ACA in unexplained RSA is controversial. In the present study, diagnostic and prognostic aspects were investigated.METHOD:One hundred five nonpregnant patients with primary, 29 with secondary RSA, and 209 controls were investigated for IgG-ACA. Follow-up studies were done during pregnancy in 76 individuals. IgM-ACA were tested in a subset of patients.RESULTS:Elevated ACA levels were significantly more frequent in both patient groups (26 and 24%) than in controls (16%). However, there was no correlation of ACA with various parameters including pregnancy outcome. In ACA-positive patients with successful pregnancy a significant decrease of ACA values during pregnancy was observed, while ACA remained high in aborting patients. IgG- and IgM-ACA correlated well.CONCLUSIONS:Although the data from nonpregnant RSA patients does not allow diagnostic or prognostic conclusions to be drawn, sequential testing of ACA-positive individuals provides the possibility to foresee pregnancy outcome.

With the growing development and routine use of high-resolution DNA typing techniques for HLA class II alleles the importance of mixed lymphocyte cultures (MLC) with respect to bone marrow transplantation is under controversial discussion. Therefore, results of MLC assays performed for donor/recipient pairs were analysed in comparison to the degree of histocompatibility as determined by prospective serological and retrospective DNA typing. Three different groups were evaluated for MLC data: (A) siblings sharing two parental haplotypes, (B) related donors sharing one haplotype, with 1-3 MM on the second haplotype, (C) phenotypically identical unrelated donors. In group A, all MLCs were negative. In contrast, 55 out of 59 MLCs in group B and 30 out of 41 in group C were positive. In group C, 11 MLCs were positive despite phenotypic class I and DRB 1 compatibility. We conclude from our results that, at present, MLC is not mandatory for genotypically HLA-identical siblings while for donor/recipient pairs of groups B and C MLC should be carried out even in cases of phenotypical compatibility for class I and DRB 1 alleles.

In the context of a controlled multicenter study on intravenous immunoglobulin (IVIG) treatment of patients with a history of unexplained recurrent spontaneous abortions (RSA), a number of controversial immunological parameters were evaluated prior to and during pregnancy with respect to their diagnostic and/or prognostic significance. A total of 390 serum samples from 52 patients were investigated. Sharing of 2 or more HLA (A, B, DR, DQ) antigens was significantly more frequent in RSA couples than in controls. The rate of cytotoxic or Fc-receptor (FcR)-blocking antibodies was not significantly lower in RSA patients than in individuals with normal pregnancies. Both tumor necrosis factor-alpha (TNF-alpha) levels and IgG anticardiolipin antibodies (IgG-ACA) were significantly increased in the patient group. While the occurrence of HLA sharing, cytotoxic/FcR-blocking antibodies and IgG-ACA did not correlate with the outcome of pregnancy, TNF-alpha levels were found to be significantly higher in patients with subsequent miscarriage than in those with successful pregnancy. IgG-ACA, if present, significantly decreased during the course of successful pregnancy but remained high in patients with subsequent abortion. It is concluded that the diagnostic and/or prognostic value of HLA sharing and cytotoxic/FcR-blocking antibodies has been overestimated while TNF-alpha and ACA levels are potential diagnostic markers and/or exhibit prognostic significance in subgroups of RSA patients.

Due to its strong "immunomodulating" effect in several well established disorders, high-dose intravenous immunoglobulins (IVIG) has been proposed as an alternative for immunotherapy with allogeneic leucocytes in patients with unexplained recurrent spontaneous abortion. This paper is intended to provide an overview on the European experience in this field.Five European pilot studies with a total of 172 patients as well as one controlled double-blind multicenter study including 64 patients were considered. In the latter, 5% human albumin was used as placebo.Success rates of the pilot studies varied from 68 to 87%. In the German controlled study, a significant specific effect of IVIG could not be verified. However, success rates for both IVIG and albumin were in the same range as for allogeneic leucocytes.At present, it is not sufficiently proven that IVIG is an appropriate tool for immunotherapy of recurrent spontaneous abortions. It is suggested that success rates of both IVIG and albumin are due to a placebo effect. However, we cannot exclude that albumin itself provides immunomodulating capacity.

For one year we determined HLA class II antigens from all patients with renal and haematological disorders and their healthy family members both by using serological methods and DNA typing (SSO, SSP). The rate of discrepancies between the results of serological DR typing and DRB1 typing was 10.8% (13/120). We were able to demonstrate that DNA typing for class II antigens leads to definite results even for patients with a poor cell quality or with lymphocytopenia where serological typing is often impossible. Furthermore, DNA typing from patients with haematological disorders is very important, especially if a bone marrow transplantation is considered. In addition to MLC testing, DRB1, DQB1 and DPB1 typing should be carried out for the patient and the potential donor. DNA typing is also recommended for patients waiting for a kidney transplantation particularly if they were serologically typed as 'homozygous', as the chance to receive an HLA-compatible organ is much higher for a heterozygous individual.

Background: Previous investigators noted an association of multiple basal cell carcinomas (BCC) with certain HLA antigens; however, these findings were contradictory, and the associations were only weak. Objective: The aim of the study was to objectify the previously found associations. Methods: Serologic HLA typing for class I and class II antigens was performed in 49 unrelated patients with 5 or more BCCs. Results: HLA-DR4 showed decreased frequencies in the patient group as compared with healthy controls (n = 716). Cw7 was found to be increased in the total group of patients as well as in a subgroup with multiple BCCs of the face (n = 24), while a subgroup with BCCs mainly on the trunk (n = 25) revealed increased frequencies of HLA-A11, -B17, -B22 and -Cw3. However, none of these deviations appeared significant after correction of p values. Conclusion: We conclude that, if at all, the HLA system plays only a minor role in the development of multiple BCCs.

Objective This study was undertaken to verify a specific effect of intravenous immunoglobulin on the outcome of pregnancy in patients with a history of recurrent miscarriage as an alternative to immunotherapy with allogeneic leucocytes.Study design In a randomised double-blind multicentre study 64 patients with a history of unexplained primary recurrent miscarriage were treated with intravenous immunoglobulin (verum) or 5% human albumin (placebo) infusions during their following pregnancy.Results Success rates for both verum and placebo, were compared excluding seven patients with explained miscarriage (20/27 (74%) versus 21/30 (70%)) and without any such exclusion (20/33 (61%) versus 21/31 (68%)). The difference between the two groups was not statistically significant.Conclusions A specific effect of intravenous immunoglobulin in primary recurrent miscarriage could not be demonstrated. These results imply the possibility of psychological influence, i.e. a placebo effect of intravenous immunoglobulin, on the outcome of pregnancy. Since success rates for both verum and placebo were in the same range as for treatment with allogeneic leucocytes, psychological effects might be responsible also for other kinds of immunotherapy for prevention of recurrent miscarriage. However, it cannot be excluded that success rates only reflect background values as reported for recurrent miscarriage patients without any treatment.

This study presents data on HLA phenotypes of 52 unrelated patients suffering from idiopathic Peyronie's disease. This first investigation on HLA class II antigens detected an association of HLA-DR3 and -DQw2 in this disorder. HLA typing was done from ACD-stabilized peripheral blood using the modified lymphocytotoxicity test. Antigen frequencies of the patient group were compared with those of healthy individuals of the local population. There were no deviations of frequencies for antigens of the B7 cross-reacting group as described in earlier studies. In addition none of the other class I antigens (HLA-A, -B, -C) showed any significant deviation in frequencies after correction of p values. Regarding class II antigens HLA-DR3 was detected in the patient group in 33.3% compared with 16.0% of the control population (corrected p < 0.05). The closely linked antigen DQw2 was found in 58.8 compared with 31.2% (corrected p < 0.005). Not only genetic factors can be stated by these findings. As HLA-DR3 and -DQw2 are known to be the typical associated antigens of organospecific autoimmune disorders, this suggests possible autoimmunological factors in this disorder of otherwise unknown etiology.

A specific effect of intravenous immune globulin (IVIG) on the outcome of pregnancy in patients with a history of habitual abortion has been postulated as an alternative to immunotherapy with allogeneic leucocytes. The results of different pilot studies have been promising, demonstrating a successful outcome of pregnancy in approximately 80% of treated patients. However, the evaluation and interpretation of the study results has to take into account that the probability of a successful pregnancy in women with a history of three spontaneous abortions is about 60% without treatment. Specific pharmacological effects therefore have to be verified in controlled studies in order to rule out psychological (placebo) effects. A specific therapeutic effect could not be verified in a German randomized, double-blind, multicentre trial in comparison to human albumin 5% which was used as a placebo. The result of another controlled study currently underway in the USA is expected.

By quantitation of elutable immunoglobulin G in a recently described ELISA, the number of HLA class I and II molecules on B-cell lines was determined using monomorphic monoclonal antibodies. An average number of 183 000 binding sites per cell for HLA class I-, 99000 for HLA-DR-, 63000 for DQ- and 42000 for DP-specific monoclonal antibodies were determined. The small amount of HLA class II molecules can in part explain the difficulties observed in HLA class II typing or in detection of class II antibodies using the lymphocytotoxicity text.