Herein, we included 527 individuals from two Hospitals, Chemnitz and University-Hospital Leipzig. In total, 199 were negative for PCR and 328 were positive upon first admission. We used next generation sequencing for HLA-A, B, C, DRB1, DRB345, DQA1, DQB1, DPA1, and DPB1, and in some cases, HLA-E, F, G, and H. Furthermore, we molecularly defined 22 blood group systems comprising 26 genes and 5 platelet antigen genes. We observed a significant enrichment of homozygosity for DQA/DQB in the positive group. Within the negative subjects, HLA-B*57:01, HLA-B*55:01, DRB1*13:01, and DRB1*01:01 were enriched, and in the positive group, homozygosity for DQA/DQB, DRB1*09:01, and DRB1*15:01 was observed. DQA1*01:01, DQA1*02:01, and DQA1*01:03 were enriched in the negative group. HLA-DQB1*06:02 was enriched in the positive group, and HLA-DQB1*05:01 and HLA-DQB1*06:03 were enriched in the negative group. For the blood group systems MNS, RH, LE, FY, JK, YT, DO, and KN, enrichment was seen in both groups, depending on the antigen under observation. Homozygosity for D-positive RHD alleles, as well as the phenotypes M-N+ of the MNS blood group system and Yk(a-) of the KN system, were enriched in the positive group. All of these significances disappeared upon correction. Subjects who carried homozygous HPA-1a were more frequent in the negative group, contrasting with the finding that HPA-1ab was enriched in the positive group.

Objective: In a cooperative study of the University Hospital Leipzig, University of Leipzig, and the Charité Berlin on kidney transplant patients, we analysed the occurrence of HLA-specific antibodies with respect to the HLA setup of the patients. We aimed at the definition of specific HLA antigens towards which the patients produced these antibodies.Methods: Patients were typed for the relevant HLA determinants using mainly the next-generation technology. Antibody screening was performed by the state-of-the-art multiplex-based technology using microspheres coupled with the respective HLA alleles of HLA class I and II determinants.Results: Patients homozygous for HLA-A*02, HLA-A*03, HLA-A*24, HLA-B*07, HLA-B*18, HLA-B*35, HLA-B*44, HLA-C*03, HLA-C*04, and HLA-C*07 in the class I group and HLA-DRB1*01, HLA-DRB1*03, HLA-DRB1*07, HLA-DRB1*15, HLA-DQA1*01, HLA-DQA1*05, HLA-DQB1*02, HLA-DQB1*03(7), HLA-DQB1*06, HLA-DPA1*01, and HLA-DPB1*04 in the class II group were found to have a significant higher antibody production compared to the heterozygous ones. In general, all HLA determinants are affected. Remarkably, HLA-A*24 homozygous patients can produce antibodies towards all HLA-A determinants, while HLA-B*18 homozygous ones make antibodies towards all HLA-B and selected HLA-A and C antigens, and are associated with an elevation of HLA-DRB1, parts of DQB1 and DPB1 alleles. Homozygosity for the HLA class II HLA-DRB1*01, and HLA-DRB1*15 seems to increase the risk for antibody responses against most of the HLA class I antigens (HLA-A, HLA-B, and HLA-C) in contrast to HLA-DQB1*03(7) where a lower risk towards few HLA-A and HLA-B alleles is found. The widely observed differential antibody response is therefore to be accounted to the patient’s HLA type.Conclusion: Homozygous patients are at risk of producing HLA-specific antibodies hampering the outcome of transplantation. Including this information on the allocation procedure might reduce antibody-mediated immune reactivity and prevent graft loss in a patient at risk, increasing the life span of the transplanted organ.

Molecular HLA typing techniques are currently undergoing a rapid evolution. While real‐time PCR is established as the standard method in tissue typing laboratories regarding allocation of solid organs, next generation sequencing (NGS) for high‐resolution HLA typing is becoming indispensable but is not yet suitable for deceased donors. By contrast, high‐resolution typing is essential for stem cell transplantation and is increasingly required for questions relating to various disease associations. In this multicentre clinical study, the TGS technique using nanopore sequencing is investigated applying NanoTYPE™ kit and NanoTYPER™ software (Omixon Biocomputing Ltd., Budapest, Hungary) regarding the concordance of the results with NGS and its practicability in diagnostic laboratories. The results of 381 samples show a concordance of 99.58% for 11 HLA loci, HLA‐A, ‐B, ‐C, ‐DRB1, ‐DRB3, ‐DRB4, ‐DRB5, ‐DQA1, ‐DQB1, ‐DPA1 and ‐DPB1. The quality control (QC) data shows a very high quality of the sequencing performed in each laboratory, 34,926 (97.15%) QC values were returned as ‘passed’, 862 (2.4%) as ‘inspect’ and 162 (0.45%) as ‘failed’. We show that an ‘inspect’ or ‘failed’ QC warning does not automatically lead to incorrect HLA typing. The advantages of nanopore sequencing are speed, flexibility, reusability of the flow cells and easy implementation in the laboratory. There are challenges, such as exon coverage and the handling of large amounts of data. Finally, nanopore sequencing presents potential for applications in basic research within the field of epigenetics and genomics and holds significance for clinical concerns.

Die Behandlung einer Prostatakrebserkrankung führt zu zum Teil langandauernden körperlichen und psychischen Beeinträchtigungen. Dennoch nimmt nur ein vergleichsweise geringer Anteil der Patienten eine Rehabilitationsmaßnahme in Anspruch; die empirische Datenlage zu den Gründen hierfür ist bislang äußerst begrenzt. Die vorliegende Studie zielte daher unter explorativer Zielsetzung auf die Identifikation von Einflussfaktoren der Inanspruchnahme einer Rehabilitationsmaßnahme bei Patienten mit einem Prostatakarzinom. Dargestellt werden die Ergebnisse des ersten Erhebungszeitpunktes einer prospektiven multizentrischen Studie zur Evaluation ambulanter onkologischer Rehabilitationsmaßnahmen. Untersucht wurden n=242 Prostatakrebspatienten in ambulanter bzw. stationärer Rehabilitation (Inanspruchnehmer), und n=253 Patienten, die keine Rehabilitation durchführten (Nichtinanspruchnehmer), mittels standardisierter Selbstbeurteilungsverfahren zu Beginn der Rehabilitationsmaßnahme bzw. nach Abschluss der Primärtherapie. Erfasst wurden folgende, aus vorliegenden Daten bei anderen Indikationsbereichen und klinischer Expertise abgeleitete unabhängige Variablen: Autonomiestreben (TPF), Selbstwirksamkeitserwartung (GSE), Distress (Distress-Thermometer), Angst und Depressivität (HADS-D), Lebensqualität (SF-8), soziale Unterstützung (SSUK), Rehabilitationsmotivation: Änderungsbereitschaft, Informationsstand, Skepsis bezüglich einer Rehabilitationsmaßnahme (PAREMO-20). Die erkrankungs- und behandlungsbezogenen Daten (Krankheitsstadium, Art und Anzahl weiterer Erkrankungen, Primärtherapien) der Patienten wurden von den behandelnden Ärzten dokumentiert. Es zeigen sich nur wenige signifikante und bedeutsame Unterschiede zwischen Rehabilitanden und Nichtinanspruchnehmern. Patienten, die keine Reha-Maßnahme durchführten, leiden häufiger unter komorbiden Erkrankungen des Verdauungssystems (18% vs. 2%, w=0,25). Beide Patientengruppen berichten deutliche Einschränkungen ihrer Lebensqualität, insbesondere in körperlicher Hinsicht. Nichtinanspruchnehmer geben signifikant höhere Werte auf der Skala „Skepsis“ des PAREMO-20 an als Rehabilitanden (eta²=0,19). Die Skala „Skepsis“ wird auch als wesentlicher Prädiktor für die Inanspruchnahme einer Rehabilitationsmaßnahme identifiziert (R²=0,23). Weitere bedeutsame Einflussfaktoren sind das Vorliegen einer zusätzlichen gastrointestinalen Erkrankung, Selbstwirksamkeitserwartung und Erwerbsstatus (R²=0,12 bzw. jeweils R²=0,02). Mit zunehmender Skepsis gegenüber dem Rehabilitationserfolg bzw. bei Komorbidität sinkt die Wahrscheinlichkeit der Reha-Inanspruchnahme. Bei Patienten mit Prostatakrebs scheint die subjektive Erwartung hinsichtlich der Effektivität einer Rehabilitationsmaßnahme zur Linderung der Beschwerden neben medizinischen Merkmalen von größerer Bedeutung für die Inanspruchnahme zu sein als soziodemografische oder belastungsbezogene Aspekte. In weiteren Studien ist zu klären, inwieweit sich dieses Ergebnis replizieren lässt, sowie die Frage, in welchem Umfang Patienten über Ziele und Behandlungskonzepte der medizinischen Rehabilitation aufgeklärt werden. Für die Praxis ist zu empfehlen, bei Information der Patienten über Rehabilitationsmaßnahmen regelhaft neben deren Belastungen auch Erwartungen und Befürchtungen zu explorieren, um so bestehende Zweifel ggf. ausräumen und die Versorgung optimieren zu können. Prostate cancer patients often suffer from treatment-associated morbidities which lead to severe physical and mental impairments. Nevertheless, only a relatively small percentage of this patient population uses medical rehabilitation services; there is still a lack of evidence concerning possible factors causing use and non-use of services. Therefore, this study exploratively aims at the identification of predictors of the use of rehabilitation services in a cohort of prostate cancer patients. In a prospective multicentre study to evaluate outpatient oncological rehabilitation services, n=242 prostate cancer patients who used outpatient or inpatient rehabilitation services (“users”) were compared with n=253 prostate cancer patients who did not use rehabilitation (“non-users”) at measure point 1. At the beginning of the rehabilitation programme and at the end of primary treatment, respectively, patients completed a self-report questionnaire consisting of standardized instruments designed to assess the following independent variables: autonomy striving (TPF), self-efficacy expectancy (GSE), distress (distress thermometer), anxiety and depression (HADS-D), quality of life (SF-8), social support (SSUK), rehabilitation motivation: readiness to change, knowledge, scepticism regarding rehabilitation services (PAREMO-20). Illness- and treatment-related variables (tumour state, comorbidity, primary therapies) were documented by the attending oncologists. Only few significant and effective differences between rehabilitation users and non-users are observed: Non-users are suffering more often from gastrointestinal comorbidities (18% vs. 2%, w=0.25). Both patient groups report severe quality-of-life impairments, particularly with respect to their physical functioning. Non-users achieve significantly higher scores on the scale “scepticism” (PAREMO-20) than users (eta²=0.19). The scale “scepticism” is also identified as the dominant predictor of utilization of rehabilitation services (R²=0.23). Suffering from comorbid diseases of the digestive system, self-efficacy expectation and employment status emerge as further significant predictors (R²=0.12, and R²=0.02 each). With increasing scepticism and in case of...

The Luminex multiplex assay for the definition of HLA antibodies was introduced 20 years ago. To date, although the assay is simple and straightforward, the interpretation of the results remains a conundrum. Making use of the so-called mean fluorescence value only does not help since this value is relative, the day-to-day variation is significantly large, and the user variation is beyond acceptable ranges. Within the same range of problems, the cutoff value used to classify HLA-specific antibodies as being either present or absent varies between laboratories. This threshold influences clinical decision-making. In addition, the readout value is used to diagnose the amount of HLA-specific antibodies after treatments like absorption, plasmapheresis, or transplantation, overlooking the relative nature of the value. In this study, we show bead variability for HLA class I and HLA class II based on negatively defined bead reactions with a mean fluorescence value below 1500 and self-antigens. In an effort to circumvent the assay’s inherent problems, we introduced the term ratio. For each serum sample, a paired sample from before the treatment is run in parallel. The ratio of post-treatment to pre-treatment can then be used to characterize antibody dynamics: a ratio above 1 reflects increased antibody production and a ratio below 1 indicates a decrease in antibodies in the serum. This value may offer the possibility to make more informed and adaptive treatment decisions.