Serology versus ARMS-PCR in prospective HLA-class I typing for bone marrow transplantation.

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, particularly 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.

Segregation of HLA genes in multicase narcolepsy families.

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.

Extensive HLA class II studies in 58 non-DRB1*15 (DR2) narcoleptic patients with cataplexy.

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 DQB1 genes, three CA repeat markers (G51152, T16CAR and G411624R) located between DQB1 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.

[Visual assessment or quantitative measurement of coronary stenoses: significance for "prima vista"-PTCA]. / Visuelle Einschätzung oder quantitative Vermessung von Koronarstenosen: Bedeutung für die "prima vista"-PTCA.

In 300 patients with 339 coronary lesions the percent diameter stenosis (%-DS) was assessed both by visual estimation and by digital quantitative coronary angiography (DQCA) by use of an on-line computer work-station. The decision for coronary angioplasty in the same setting ("prima vista"-PTCA) was based on history, evidence of ischemia and visual estimation of %-DS. DQCA measurements of the 339 stenoses revealed a normal distribution of lesion severity with a mean of 58.4 +/- 11.3%. In contrast to DQCA visual estimation led to a bimodal distribution with a nadir at approximately 55% between two peaks at approximately 45% and approximately 75% and a mean of 70.5 +/- 19.6%. Visual estimation underestimated lesions in the range of 30-55% and overestimated the %-DS between 55-99%. Visual estimation revealed a %-DS > or = 60% in 251 stenoses (74.0%) of the 339 lesions, an estimate that led to subsequent "prima vista"-PTCA. Conversely, DQCA revealed only 184 stenoses (54.3%) with a %-DS > or = 60%; thus, 86 stenoses (25.3%) did not meet the morphologic indication criteria for PTCA. The bimodal distribution of stenosis severity according to visual analysis with an overestimation of borderline stenosis severity reflects at tendency for "self-referral" of patients for PTCA. DQCA serves as an objective tool in the decision-making process for PTCA and may reduce "cosmetic" interventions or justify to defer PTCA. Especially in the selection process for "prima vista"-PTCA DQCA-quantification of stenosis severity is recommended.

On-site digital quantitative coronary angiography: comparison with visual readings in interventional procedures. Implications for decision and quality control.

In order to review the morphological criterion for an interventional procedure, diameter stenosis (%DS) of 226 coronary lesions in 200 patients undergoing elective coronary angiography with an option for 'prima vista' angioplasty (pPTCA), was assessed on-site by both visual 'eye balling' (EB) and independent digital quantitative coronary angiography (DQCA) by means of an angiographic workstation. Compared to DQCA, EB overestimated the %DS between 50 and 80% and accounted for the majority of discrepancies with overestimation up to 45%. Concordant estimates of %DS by both methods were observed in only 10 of the total of 226 stenotic segments; in 20 of 226 cases, EB underestimated %DS up to 20%. EB revealed a %DS > or = 60% in 166 stenoses (73.4%), an estimate that led to subsequent pPTCA. However, only 119 (52.6%) of these lesions had a %DS > or = 60% as assessed objectively by DQCA. With regard to the criterion for PTCA 47 of 166 performed pPTCA (28.3%) would not meet the indication criteria based on objective DQCA information. EB and DQCA (+/-5%DS) had concordant results and criteria for pPTCA only in 103 of 166 coronary lesions (62.1%). These results lead to the conclusion that, on-site and on-line DQCA by an independent cardiologist eliminates both under- and overestimation of stenoses as seen with EB. DQCA supports immediate decision-making and appears necessary for reliable evaluation of coronary morphology in an interventional catheterization laboratory setting and may eventually ensure intraprocedural quality control.

Differences in lipoprotein (a) and apolipoprotein (a) levels in men and women with advanced coronary atherosclerosis.

BACKGROUND: This study was designed to evaluate whether differences between sexes exist in serum-lipoprotein (a) [Lp(a)] and arterial-wall-apolipoprotein (a) [Apo(a)] levels in patients with advanced coronary artery disease. METHODS: The concentrations of Lp(a) in serum and Apo(a) in aortic biopsies were studied in 76 men and 20 women undergoing coronary artery bypass graft surgery. The severity of coronary artery disease was determined by a coronary atherosclerosis score that used quantitative coronary angiography. RESULTS: Serum-Lp(a) and tissue-Apo(a) do not correlate with the severity of coronary artery disease as expressed by the coronary atherosclerosis score (r = 0.09 and r = 0.14, respectively). Women were older (65 +/- 8 versus 57 +/- 8 years, P < 0.001) and had higher mean Lp(a) and higher mean Apo(a) levels (47 +/- 41 versus 32 +/- 40 mg/dl and 33 +/- 34 versus 19 +/- 24 micrograms/g wet weight, P < 0.05) than men with identical coronary atherosclerosis score (35 +/- 8 versus 33 +/- 8, P > 0.05). The serum levels of cholesterol, triglycerides, and high-density lipoprotein were similar in both groups. CONCLUSIONS: Men and women undergoing coronary artery bypass graft surgery had very similar severity of coronary artery disease as expressed by the coronary atherosclerosis score. Women were 8 years older and had 1.5 times higher mean serum-Lp(a) levels and 1.75 times higher mean tissue Apo(a) levels higher than the men. Sixty per cent of the women but only 39% of the men had serum Lp(a) levels higher than 25 mg/dl. Lp(a) level seems to be an additional risk factor for coronary artery disease confined to postmenopausal women.

Diagnostic and prognostic significance of anticardiolipin antibodies in patients with recurrent spontaneous abortions.

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.

[Association of lipoprotein (a) with the extent of coronary atherosclerosis in surgically revascularized men and women]. / Assoziation von Lipoprotein (a) mit dem Ausmass der koronaren Atherosklerose bei chirurgisch revaskularisierten Männern und Frauen.

Lipoprotein (a) (Lp(a)) levels are genetically determined and levels higher than 25 mg/dl are associated with increased prevalence of coronary artery disease (CAD). We studied gender differences in 76 men and 20 women undergoing coronary artery bypass graft surgery (CABG) for a potential association between Lp(a) levels both in serum and the aortic wall (Apo(a)) and the severity of CAD determined by an atherosclerosis score (CS) using quantitative coronary angiography (QCA). Serum Lp(a) and tissue Apo(a) do not correlate with the severity of CAD as assessed from QCA (r = 0.09 and r = 0.14, resp.). 60% of women but only 39% of men had serum Lp(a) levels higher than 25 mg/dl. Women were 8 years older (65 +/- 8 vs. 57 +/- 8 years, p < 0.001) and had 1.5 times higher mean serum Lp(a) and 1.75 times higher mean tissue Apo(a) levels (47 +/- 41 vs. 32 +/- 40 mg/dl and 33 +/- 34 vs. 19 +/- 24 micrograms/g WW, p < 0.05) than men with identical CS (35 +/- 8 vs. 33 +/- 8, p = NS). The serum levels of cholesterol, triglycerides, and high-density lipoprotein were similar in the two groups. There is no association between Lp(a) and Apo(a) and the severity of coronary atherosclerosis in men and women undergoing coronary artery bypass surgery.

Immunogenetic and serological investigations in nonpregnant and in pregnant women with a history of recurrent spontaneous abortions. German RSA/IVIG Study Group.

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.

[New possibilities of donor selection for bone marrow and organ transplantation by HLA typing and genomic DNA]. / Neue Möglichkeiten der Spenderauswahl für die Knochenmark- und Organtransplantation durch HLA-Typisierung aus genomischer DNA.

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.