Class I typing by PCR-SSP in Olomouc.

In this article, methodical experience and population data are given for application of PCR-SSP/ARMS method (Phototyping) to HLA class I typing in Olomouc. Phototyping is reliable method suitable for typing of small to medium number of samples. Method is fast enough for use in on-call service, resolution is better than the level of good serology, and price of method is comparable with serology. Our experience and tips are described below. Population data of healthy unrelated individuals for HLA-A, -B, -Cw are given in the tables.

Familial hydronephrosis unlinked to the HLA complex.

Clinical findings, management, and possible linkage of congenital hydronephrosis caused by ureteropelvic junction stenosis to the HLA complex were studied in four families. These families provide evidence of possible autosomal dominant inheritance. HLA class I antigen studies in all four and class II (HLA-DR) in three families were performed. These studies failed to show close linkage to the chromosome 6 markers in two families but there was consistent inheritance in the other two. Although formal linkage calculations are not presented, it is apparent that in some families HLA haplotyping is not useful in predicting prevence of renal obstruction.

"The sarcoidosis map": a joint survey of clinical and immunogenetic findings in two European countries.

We pooled immunogenetic data obtained in independent studies in two European populations (Italian and Czech) of patients affected by sarcoidosis. Correspondence analysis was used to investigate the associations between clinical and immunogenetic data. Two hundred and thirty-three patients were enrolled in the study, of which 126 were from the Czech Republic and 107 from Italy. Using a common protocol, we examined each patient for sex, age of disease onset, roentgenologic stage, extrapulmonary spread, and clinical course. One thousand and ten healthy individuals, HLA typed for class I and II serologic polymorphisms, served as controls. Findings that were essentially in agreement in both populations were: (1) a positive association of sarcoidosis with HLA-A1, B8, and DR3 markers, and a negative association with HLA-B12 and DR4; (2) a prevalence of HLA-DR3 and DR4 among females and of DR5 among males; (3) a relationship of B13 and B35 with early onset and of A30, B8, DR3, and DR4 with late onset of disease; (4) an association of B27 with sarcoidosis restricted to the lungs; (5) a relationship of A1, B8, B27, and DR3 to roentgenologic stage I and of B12 and DR4 to stage III; and (6) an association of HLA-DR3 with a good outcome. Population-restricted findings essentially concerned the alleles HLA-B13 and B22, the former being associated with the disease, male sex, early onset, extrapulmonary localization and relapse only in Czechs, and the latter to disease spread only in Italians. Our results seem to support the concept that immunogenetic background may at least partly account for the clinical heterogeneity of sarcoidosis.

[The central registry of bone marrow donors and the program for bone marrow transplantation from unrelated donors--20 months' experience]. / Centrální registr dárcu kostní drene a program nepríbuzenských transplantací kostní drene--dvacetimesícní zkusenosti.

The authors describe their 20 months' experience with the establishment of a bone marrow register in the Czech Republic and its practical association with the programme of bone marrow transplantations from non-related subjects. During the 20 months of activity 5455 voluntary bone marrow donors were examined and at present more than 500 new voluntary donors are examined every month. For 21 patients a HLA identical unrelated donor is sought. For four patients a HLA compatible donor was found. The first bone marrow transplantation from a unrelated donor was performed and another patient is prepared for transplantation within the next few days. The authors discuss professional, organizational, moral and ethical problems associated with the elaboration of this medical programme.

Polymorphism of the 5' flanking region of the HLA-DQA1 gene in coeliac disease.

Coeliac disease (CD) is associated with particular HLA genotypes. The susceptibility gene (or genes) has been mapped to the class II region, most probably to the DQ loci. Polymorphism of the upstream promoter region of the DQA1 gene (QAP) has been recently reported. At least ten variants or QAP alleles have been found, some of which are present in the cis-acting regulatory sequences. Allelic differences in DQ molecule expression may play a role in susceptibility to CD. We investigated the QAP polymorphism in 102 CD patients and 142 unrelated healthy controls of Czech origin using polymerase chain reaction amplification (PCR) of genomic DNA and oligonucleotide probes. We found a significant frequency increase of the alleles QAP 4.1 (RR = 10.3, p.c. = 10(-6) and QAP 2.1 (RR = 2.4, p.c. = 0.017) in patients over controls. An increased susceptibility is provided by the presence of both alleles, as is shown by the higher proportion of QAP 4.1, 2.1 heterozygotes among patients than expected from the Hardy-Weinberg equilibrium and by the comparison of the odds ratios for these alleles. There is a strong linkage disequilibrium between the QAP alleles and the DQA1, DQB1, and DRB1 loci. Two haplotypes carrying the QAP alleles whose frequency is increased are predominant in this group of CD patients: DQB1*0201, DQA1*0501, QAP4.1, DRB1*0301 and DQB1*0201, DQA1*0201, QAP 2.1, DRB1* 0701. Thus, the QAP variants are increased as part of these haplotypes and we cannot discriminate if they are responsible for the primary association.

[HLA antigens in patients with celiac disease]. / Výskyt HLA antigenu u nemocných s celiakií.

In 104 non-related children with coeliac disease, all from northern Moravia, the authors assessed the distribution of HLA antigens class I and II and compared the findings with a control group. They revealed a positive association between HLA A1 (62% sick subjects as compared 27% healthy ones), B8 (68% as compared with 18%), DR3 (66% as compared with 14%) and DQw2 (79% as compared with 23%). In HLA antigens class II there is a higher relative risk of the disease for subjects with antigens DR3 and DQw2 and a higher aetiological fraction for the investigated antigens.

Quantitative indices in paternity cases.

The study discusses the basic quantitative indices used as a standard method in foreign professional literature dealing with paternity cases. They are as follows: 1. mean probability of exclusion (PE) which characterizes the informative value of the experts opinions and is the same in all the disputes evaluated by this expert. 2. relative frequency of men chosen at random from the population and excluded at given phenotype of mother and child (RME). 3. probability of paternity (PP) for particular trio: mother-child-the accused man. Hereinafter the results of our studies in the HLA laboratory in Olomouc from 1976-1991 are introduced.

HLA-A and HLA-B typing: classification of errors.

The authors followed the frequency of errors in serologic identification of HLA-A and HLA-B antigens. They analyzed 205 paternity cases evaluated independently by two judicial experts (a revision of expert opinions). The specification of revealed discrepancies is demonstrated in the study and possible causes of their origin are analyzed. In the conclusions the authors recommend useful steps leading to the decrease of error frequency in HLA antigen typing. The HLA system--the major histocompatibility system of a man, has a broad utilization in the field of practical medicine. HLA compatibility should be respected in the choice of the couple donor-recipient for transplant therapy. HLA, as a highly polymorphic system, is successfully applied in forensic medicine. A strong association with some diseases makes it possible to utilize HLA as a supporting diagnostic sign. The commission for nomenclature of WHO at XI. International Histocompatibility Workshop (1991) recognized the following numbers of HLA specificities: 25 A, 55 B, 10 C, 21 DR, 9 PQ, 6 DP. At present HLA laboratories used mostly serological methods. The need to mutually differentiate HLA antigens which are similar due to their molecular structure puts an extraordinary demand on HLA laboratories. On one hand, a great number of HLA phenotypes exist in the population, which ensures the identification of every individual and the distinguishing of self-nonself components. On the other hand there appear the limited possibilities of HLA laboratories with relatively low numbers of diagnostic sera. We inquired how often errors occurred in HLA antigens determination. We chose a set of 205 auditing experts opinions of paternity cases for analyzing the problem.(ABSTRACT TRUNCATED AT 250 WORDS)

HLA antibodies in renal transplant candidates.

The level of anti-HLA antibodies was followed in 155 patients of the chronic dialyzed program: against antigens HLA Class I and antigens HLA-DR representing HLA Class II. Correlation between the titre of these antibodies and additional factors were analyzed. The effect of pregnancy and consequently of sex was proved. Of significance were the number of dialyses undergone and the time of inclusion into the dialyses program. Positive correlation between the titre of both types of anti-HLA antibodies is so significant that it makes possible to concentrate on the more simple anti HLA I antibodies determination. The result of crossmatch test is of crucial importance when making decisions on suitable recipient of the kidney from the set of patients of dialyzed program (DP). The presence of anti-HLA alloantibodies can cause hyperacute rejection. The anti-HLA alloantibodies titre is regularly followed in DP patients. Several significant hypotheses can be verified in a large enough set of patients (hypotheses marked with ordinal numbers 1-5): 1. Only the titre of anti-HLA Class I antibodies is usually followed in DP patients. Some workplaces abroad consider it useful to follow even the anti-HLA Class II antibodies. Is anti-HLA I and HLA II antibodies titre in such a significant correlation that it is sufficient to follow only anti-HLA Class I? 2. Mechanisms of HLA antigen sensitization in DP patients are well known. We can verify the influence of transfusion number, allograft rejections and number of previous pregnancies on the anti-HLA antibodies titre.(ABSTRACT TRUNCATED AT 250 WORDS)

HLA antigens associated with sarcoidosis.

In a series of 123 sarcoidosis patients (inhabitants of Moravia), frequencies of 15 HLA-A, 31 HLA-B, and 7 HLA-C antigens have been found. (Control group consisted of 500 healthy persons from the same region.) A subgroup of 46 patients was examined in order to determine a frequency of 10 HLA-DR antigens. (Control group consisted of 146 persons.) A positive association was proved between sarcoidosis and the HLA-B8 and B13 antigens (RR = 2.8 and RR = 3.1, respectively). A frequency of B8B13 heterozygotes was highly significant (RR = 8.5). The observed antigen genotype frequencies may be explained by the hypothesis of 2 HLA-linked disposing genes.