Immunisation against Human Leucocyte Antigens (HLA) can be caused by pregnancy, blood transfusion, or organ transplants. The HLA antibody status of a given patient significantly influences their access and waiting time to transplant. For some highly sensitised patients (HSP) there is hardly any suitable donor available in the deceased donor pool of their allocation organisation and therefore they wait a very long time before being offered a kidney for transplant. Especially patients with rare HLA phenotypes in relation to the actual donor pool are waiting extremely long. As HLA phenotypes are different in the various European populations, we hypothesized that extension of the donor pool outside the respective allocation system will increase the chance of receiving a compatible transplant for this subgroup of highly sensitised patients. One of the objectives of the EUROSTAM project, (a Europe-wide Strategy to enhance Transplantation of highly sensitised patients on the basis of Acceptable HLA Mismatches) was to develop a tool to compare the chance of transplanting HSP in different European populations with donor organs from within and outside their own donor pool. Information on the HLA type and ABO blood group of the actual donor population, as well as the acceptable mismatches of long waiting HSP were obtained from the EUROSTAM partner organizations i.e. Eurotransplant (ET), UK National Health Service Blood and Transplant (NHSBT), Barcelona, Prague and Athens. Results from simulations using the newly developed tool shows that 195 (27%) of the 724 long waiting highly sensitised patients registered at each partner organisation have increased chances of transplant in a different European donor pool. This makes a strong case for sharing kidneys between European countries for selected difficult to transplant patients.

• Klíčová slova
• Acceptable mismatch, HLA, Highly sensitised patient, Kidney, Transplantation,
• MeSH
• dárci tkání MeSH
• histokompatibilita MeSH
• HLA antigeny genetika   imunologie   MeSH
• imunizace MeSH
• lidé MeSH
• příjemce transplantátu MeSH
• seznamy čekatelů MeSH
• testování histokompatibility metody   MeSH
• transplantace ledvin * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• úvodníky MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• HLA antigeny MeSH

Extended molecular characterization of HLA genes in the IHWG reference B-lymphoblastoid cell lines (B-LCLs) was one of the major goals for the 17th International HLA and Immunogenetics Workshop (IHIW). Although reference B-LCLs have been examined extensively in previous workshops complete high-resolution typing was not completed for all the classical class I and class II HLA genes. To address this, we conducted a single-blind study where select panels of B-LCL genomic DNA samples were distributed to multiple laboratories for HLA genotyping by next-generation sequencing methods. Identical cell panels comprised of 24 and 346 samples were distributed and typed by at least four laboratories in order to derive accurate consensus HLA genotypes. Overall concordance rates calculated at both 2- and 4-field allele-level resolutions ranged from 90.4% to 100%. Concordance for the class I genes ranged from 91.7 to 100%, whereas concordance for class II genes was variable; the lowest observed at HLA-DRB3 (84.2%). At the maximum allele-resolution 78 B-LCLs were defined as homozygous for all 11 loci. We identified 11 novel exon polymorphisms in the entire cell panel. A comparison of the B-LCLs NGS HLA genotypes with the HLA genotypes catalogued in the IPD-IMGT/HLA Database Cell Repository, revealed an overall allele match at 68.4%. Typing discrepancies between the two datasets were mostly due to the lower-resolution historical typing methods resulting in incomplete HLA genotypes for some samples listed in the IPD-IMGT/HLA Database Cell Repository. Our approach of multiple-laboratory NGS HLA typing of the B-LCLs has provided accurate genotyping data. The data generated by the tremendous collaborative efforts of the 17th IHIW participants is useful for updating the current cell and sequence databases and will be a valuable resource for future studies.

• Klíčová slova
• B-lymphoblastoid cell lines, Human leukocyte antigen, International HLA and Immunogenetics Workshop, Multiple-laboratory testing, Next-generation sequencing,
• MeSH
• alely MeSH
• B-lymfocyty virologie   MeSH
• exony genetika   MeSH
• genetická variace MeSH
• genetické lokusy MeSH
• genotyp MeSH
• haplotypy genetika   MeSH
• histokompatibilita - antigeny třídy I genetika   MeSH
• histokompatibilita - antigeny třídy II genetika   MeSH
• histokompatibilita MeSH
• HLA antigeny genetika   MeSH
• homozygot MeSH
• jednoduchá slepá metoda MeSH
• lidé MeSH
• sekvenční analýza DNA metody   MeSH
• správnost dat MeSH
• testování histokompatibility metody   MeSH
• transformované buněčné linie MeSH
• virová transformace buněk MeSH
• virus Epsteinův-Barrové imunologie   MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• histokompatibilita - antigeny třídy I MeSH
• histokompatibilita - antigeny třídy II MeSH
• HLA antigeny MeSH

The 17th International HLA and Immunogenetics Workshop (IHIW) organizers conducted a Pilot Study (PS) in which 13 laboratories (15 groups) participated to assess the performance of the various sequencing library preparation protocols, NGS platforms and software in use prior to the workshop. The organizers sent 50 cell lines to each of the 15 groups, scored the 15 independently generated sets of NGS HLA genotyping data, and generated "consensus" HLA genotypes for each of the 50 cell lines. Proficiency Testing (PT) was subsequently organized using four sets of 24 cell lines, selected from 48 of 50 PS cell lines, to validate the quality of NGS HLA typing data from the 34 participating IHIW laboratories. Completion of the PT program with a minimum score of 95% concordance at the HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 loci satisfied the requirements to submit NGS HLA typing data for the 17th IHIW projects. Together, these PS and PT efforts constituted the 17th IHIW Quality Control project. Overall PT concordance rates for HLA-A, HLA-B, HLA-C, HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DQB1, HLA-DRB1, HLA-DRB3, HLA-DRB4 and HLA-DRB5 were 98.1%, 97.0% and 98.1%, 99.0%, 98.6%, 98.8%, 97.6%, 96.0%, 99.1%, 90.0% and 91.7%, respectively. Across all loci, the majority of the discordance was due to allele dropout. The high cost of NGS HLA genotyping per experiment likely prevented the retyping of initially failed HLA loci. Despite the high HLA genotype concordance rates of the software, there remains room for improvement in the assembly of more accurate consensus DNA sequences by NGS HLA genotyping software.

• Klíčová slova
• NGS HLA typing, Proficiency testing, Quality control, Reference cell panel,
• MeSH
• alely MeSH
• genotyp * MeSH
• HLA antigeny genetika   MeSH
• imunogenetika * MeSH
• konsensuální konference jako téma MeSH
• lidé MeSH
• mezinárodní spolupráce MeSH
• pilotní projekty MeSH
• řízení kvality MeSH
• software MeSH
• testování histokompatibility metody   MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• HLA antigeny MeSH

The aim of our study was to evaluate the relevance of complement-binding donor-specific antibodies (DSA) for prediction of antibody-mediated rejection (AMR) after liver transplantation. Sera from 123 liver transplant recipients were retrospectively defined for HLA specificity and complement-fixing activity using the single antigen beads, C1q and C3d techniques. Liver-recipients' sera were tested before transplantation, 3, 6 months and 1 year after transplantation. Patients were followed up for graft survival and rejection incidence for 1 year after transplantation. All patients with pretransplant complement-binding DSA developed severe AMR after transplantation, while three recipients out of four, who produced de novo complement-fixing DSA, developed AMR. Definition of DSA with respect to complement-fixing activity may provide clinically relevant information about the risk of AMR after liver transplantation.

• Klíčová slova
• C4d, HLA, antibodies, complement, crossmatch, liver transplantation, rejection,
• MeSH
• dárci tkání MeSH
• dítě MeSH
• dospělí MeSH
• homologní transplantace MeSH
• isoprotilátky krev   MeSH
• komplement C1q metabolismus   MeSH
• komplement C3d metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• následné studie MeSH
• přežívání štěpu * MeSH
• prognóza MeSH
• rejekce štěpu krev   diagnóza   imunologie   patologie   MeSH
• retrospektivní studie MeSH
• senioři MeSH
• testování histokompatibility metody   MeSH
• transplantace jater * MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• isoprotilátky MeSH
• komplement C1q MeSH
• komplement C3d MeSH

A novel therapeutic approach to refractory acute antibody-mediated rejection (AMR) in kidney transplant recipients was applied in 23 patients based on administration of Bortezomib, intravenous corticosteroids, plasmapheresis and Rituximab. Application of Bortezomib regimen led to diminishing of donor-specific antibodies (DSA) to HLA-B (P = 0.004) and HLA-DR (P = 0.0005), but not to HLA-A (P = 0.106) and HLA-DQ antigens (P = 0.18). Patients with good clinical response to treatment had significantly better allograft survival than recipients with continuing deterioration of graft function (P = 0.019). Graft survival after therapy of refractory AMR was significantly worse than survival after first transplantation and was comparable with outcomes after retransplantation. In conclusion, therapy with Bortezomib was well tolerated and effective in decreasing the levels of HLA-B and -DR antibodies, however, was not successful in depleting HLA-A and -DQ DSA.

• Klíčová slova
• Bortezomib, antibody-mediated rejection, donor-specific antibodies, intravenous immunoglobulins, plasmapheresis,
• MeSH
• bezpečnost pacientů MeSH
• bortezomib terapeutické užití   MeSH
• dárci tkání MeSH
• dospělí MeSH
• HLA antigeny genetika   imunologie   MeSH
• homologní transplantace MeSH
• hormony kůry nadledvin terapeutické užití   MeSH
• imunologické faktory terapeutické užití   MeSH
• isoprotilátky krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• pilotní projekty MeSH
• plazmaferéza metody   MeSH
• přežívání štěpu * MeSH
• prognóza MeSH
• rejekce štěpu krev   diagnóza   imunologie   patologie   MeSH
• rituximab terapeutické užití   MeSH
• rozvrh dávkování léků MeSH
• testování histokompatibility metody   MeSH
• transplantace ledvin * MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bortezomib MeSH
• HLA antigeny MeSH
• hormony kůry nadledvin MeSH
• imunologické faktory MeSH
• isoprotilátky MeSH
• rituximab MeSH

BACKGROUND: The selection of optimal donor is crucial for successful hematopoietic stem cell transplantation (HSCT). Thereby, it is appropriate to know, in addition to basic human leukocyte antigen (HLA) gene matches, other immunogenic or nonimmunogenic parameters predicting the outcome of transplant. OBJECTIVE: A unified approach is necessary to provide a comprehensive view of the patient-donor compatibility characterization outside of standard HLA genes. The approach should be applicable as a tool for optimizing procedures for extended donor typing and/or verification typing of a donor. METHODS: The study used the summary, unification, and innovation of existing practical knowledge and experience of the Czech National Marrow Donor Registry of various factors beyond HLA matching with impact on transplant outcome. RESULTS: An information technology system-implemented procedure (a verification algorithm) is presented as the decision support approach for prematurely discarding less suitable donors from the transplantation process. It is intended primarily for the transplant specialist to help establish optimal procedures for verifying and determining donor critical factors. CONCLUSIONS: A process defining HLAs, killer cell immunoglobulin-like receptors, and cytokine typing strategies was proposed to provide support to a transplant specialist in refining the choice of a suitable donor.

• MeSH
• algoritmy * MeSH
• HLA antigeny imunologie   MeSH
• lidé MeSH
• receptory KIR imunologie   MeSH
• registrace MeSH
• systémy pro podporu klinického rozhodování * MeSH
• testování histokompatibility metody   MeSH
• transplantace hematopoetických kmenových buněk metody   MeSH
• výběr dárců metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Československo MeSH
• Názvy látek
• HLA antigeny MeSH
• receptory KIR MeSH

BACKGROUND: The association of donor-specific HLA antibodies (DSA) with kidney graft failure has been addressed previously; however, the majority of studies were based on small numbers of patients with graft failure. METHODS: We investigated 83 patients with failed kidney transplants for a possible association of de novo development and persistence or loss of pre-existing DSA with graft failure. Single Antigen Bead assay-detected DSA and non-DSA antibodies were compared between patients with graft loss and matched controls with functioning grafts. RESULTS: The incidence of weak de novo DSA or non-DSA at a mean fluorescence intensity of 500 or higher was higher in the graft loss than in the nonrejector group (76% vs 40%, P < 0.001). Because of the low number of patients developing de novo DSA, the DSA results did not reach statistical significance (only 22% of patients with graft loss developed de novo DSA). However, at all cutoffs, there was a significantly higher rate of graft loss in patients with de novo non-DSA. The incidence of strong pretransplant DSA that persist after transplantation was higher in the graft loss group (10% vs 1%, P = 0.034). When C1q-binding ability in sera of rejectors and nonrejectors with posttransplant de novo or persistent DSA was compared, none of the nonrejectors demonstrated C1q positivity, whereas 43% of patients with graft loss showed C1q-positive antibodies, although not necessarily donor-specific (P < 0.001). CONCLUSIONS: Our data show that the posttransplant presence of persisting or de novo HLA antibodies, especially if C1q binding, is associated with graft loss, even if the antibodies are not specific for mismatched donor HLA.

• MeSH
• biologické markery krev   MeSH
• dospělí MeSH
• histokompatibilita MeSH
• HLA antigeny imunologie   MeSH
• isoprotilátky krev   MeSH
• komplement C1q imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• prediktivní hodnota testů MeSH
• rejekce štěpu krev   diagnóza   imunologie   MeSH
• retrospektivní studie MeSH
• rizikové faktory MeSH
• senioři MeSH
• sérologické testy MeSH
• testování histokompatibility metody   MeSH
• transplantace ledvin škodlivé účinky   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• HLA antigeny MeSH
• isoprotilátky MeSH
• komplement C1q MeSH

AIMS: To determine the impact of HLA compatibility measured by the Compatibility Index, on survival, rate of rejections, malignancies and infections in patients after heart transplantation (HTx). METHODS: We carried out a retrospective analysis of 182 consecutive patients who underwent heart transplantation in our center from January 2001 to April 2010. According to degree of HLA-A, B and DR matching (Compatibility Index, CI) the patients were divided in two groups, Group A (n=83) with an IC 0-17 and group B (n=99) with an IC 18-26. There was no significant difference in demographic parameters between recipients and donors. RESULTS: We found no difference in rates of rejections or infections between Group A and Group B (AR: 22 (26.5%) vs. 34 (34.3%), P=0.2539; infections: 21 (25.3%) vs. 27 (27%) P=0.7637). The distribution of infections in terms of type (bacterial, viral, fungal, including Aspergillus) was similar in both groups. The incidence of malignant tumours was infrequent (3 (3.6%) vs. 4 (4.0%), P=0.8817). We found trend toward lower level of tacrolimus in Group A. Long term survival was similar in both groups. CONCLUSIONS: Based on the results of our single-center trial, we found no impact of higher degree of HLA-A,-B, and -DR matching on survival, rejection episodes or infection. Further large studies are necessary to confirm our hypothesis that subjects with better HLA compatibility could require lower dose immunosuppression.

• MeSH
• biologické markery metabolismus   MeSH
• dospělí MeSH
• HLA antigeny imunologie   MeSH
• HLA-A antigeny imunologie   MeSH
• HLA-B antigeny imunologie   MeSH
• HLA-DR antigeny imunologie   MeSH
• hodnocení rizik MeSH
• imunosupresiva terapeutické užití   MeSH
• Kaplanův-Meierův odhad MeSH
• lidé středního věku MeSH
• lidé MeSH
• následné studie MeSH
• přežívání štěpu imunologie   MeSH
• rejekce štěpu imunologie   MeSH
• retrospektivní studie MeSH
• rizikové faktory MeSH
• senioři MeSH
• testování histokompatibility * metody   MeSH
• transplantace srdce * metody   mortalita   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• biologické markery MeSH
• HLA antigeny MeSH
• HLA-A antigeny MeSH
• HLA-B antigeny MeSH
• HLA-DR antigeny MeSH
• imunosupresiva MeSH

The HLA complex is the most polymorphic genetic system in man yet known. The variability of the HLA antigens is given by the presence of many alleles of the HLA genes. Requirements for compatibility of HLA antigens in organ and bone marrow transplantations, and also in the determination of genetic risk factors in autoimmune diseases evoke strong pressure on progress in HLA typing methods, mainly for increasing their sensitivity and resolution. For typing of the HLA antigens there are used cellular, serological, biochemical and DNA methods. For HLA class I typing there following tests are used: cytotoxic test (serological), CML (cellular), 1D--IEF (biochemical), RFLP, SSO, SSP--PCR, and SBT (DNA methods). For HLA class II typing, cytotoxic test (cellular), MLC (serological), RFLP, SSO, SSP--PCR, and SBT (DNA methods) are used. DNA methods represent the modern trend in the area of HLA typing and it will probably replace larger part of other HLA typing techniques. In our article, we describe the principles of methods that are used for HLA typing.

• MeSH
• HLA antigeny genetika   MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• polymorfismus genetický * MeSH
• testování histokompatibility metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• HLA antigeny MeSH

DNA typing for HLA class H presents quality typing which improves kidney graft survival. Discrepancy between serological and molecular biological typing of major histocompatibility complex varies from ten to fifteen percent according to the European Tissue Typing Laboratories. In conditions of Regional Tissue Typing Laboratory we started with molecular HLA-DR typing in the beginning of 1997. One of the reasons was large activity of Regional Transplantation Centre in University Hospital and as we mentioned before quality of molecular HLA typing. Till November 1997 we took and typed 43 donors organs and we transplanted about 57 kidneys and 3 livers. That was why to introduce molecular biological methods of HLA-DR typing into the practical life of laboratory especially on duties seemed to be very advisable. We compare serological and PCR-SSP methods in group of 30 cadaveric donors in this article. The age median of our group was 36.5 yrs, the ratio men-women was 20:10.

• MeSH
• dárci tkání * MeSH
• dospělí MeSH
• geny MHC třídy II genetika   MeSH
• HLA-DR4 antigen genetika   MeSH
• HLA-DR7 antigen genetika   MeSH
• laboratoře * MeSH
• lidé MeSH
• mrtvola MeSH
• polymerázová řetězová reakce metody   MeSH
• senzitivita a specificita MeSH
• testování histokompatibility metody   MeSH
• transplantace jater MeSH
• transplantace ledvin MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• HLA-DR4 antigen MeSH
• HLA-DR7 antigen MeSH