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Background and Objectives: In Germany, the donor history questionnaire (DHQ) is traditionally filled in at the donation center to avoid any influence of others. Since March 2020, it has been suggested to donors to answer the DHQ already at home and to call if they have any concerns to reduce the number of ineligible donors on-site during the COVID-19 pandemic. Materials and Methods: We evaluated the rate of ineligible donors before and after March 2020. Additionally, an anonymous online survey asking for the donors’ attitude towards the DHQ was performed. It included questions on whether and for what reason the DHQ had been answered incorrectly in the past. Results: The rate of ineligible donors decreased by 27% (from 7.1% to 5.2%). In total, 5,556 of 10,252 invited donors completed the survey (54.2%). 88.6% reported either going through the DHQ at home or knowing all questions from their previous donations. 444 donors (8.0%) had at least once postponed a donation after reading the DHQ at home. 68 donors (1.2%) admitted having intentionally provided false answers in the past (9 at home, 43 on-site, 14 both, 2 unknown). Not wanting to be rejected once arriving at the donation center was an important motivation for 42% of donors answering incorrectly on-site. Details on 46 incorrect answers were provided: only 17 had no influence on donor eligibility or product quality. In 5 cases, some blood products might have had impaired quality. Truthful answers to 17 questions would have led to deferral, mostly due to increased risk for unrecognized viral infections transmitted by sexual contacts. For a further 7 questions, there was insufficient information available to determine possible consequences. Asked about their general opinion, 753 (13.6%) of all donors estimated the risk of incorrect answers being greater on-site, while 239 (4.3%) presumed an increased risk at home. Conclusion: Answering the DHQ prior to a donation visit prevented ineligible donors from visiting the donation center. Furthermore, it might improve honesty, as the discomfort of being deferred after arriving at the donation center was an important reason to answer incorrectly. Overall, there was no increased risk of donor or product safety, and potentially even a benefit.

Introduction The most frequently reported neurological adverse event of ChAdOx1 nCoV-19 (AZD1222) vaccine is headache in 57.5%. Several cases of cerebral venous thrombosis (CVT) have developed after vaccination. Headache is the leading symptom of CVT. For the differential diagnosis of headaches attributed to this vaccine and headaches attributed to CVT, it is of central clinical importance whether and, if so, how the phenotypes and course of these headaches can be differentiated. The study aims to examine in detail the phenotype of headache attributed to this vaccine. Methods Data on the clinical features and corresponding variables were recorded using a standardized online questionnaire in this multicenter observational cohort study. The primary outcomes of this study are the clinical features of headaches after vaccination. Findings A total of 2464 participants reported headaches after vaccination with the ChAdOx1 nCoV-19 (AZD1222) vaccine. On average, headaches occurred 14.5 ± 21.6 h after vaccination and lasted 16.3 ± 30.4 h. A bilateral location was described by 75.8% of participants. This is most often found on the forehead (40.0%) and temples (31.4%); 50.4% reported a pressing and 37.7% a dull pain character. Headache intensity was most often severe (38.7%), moderate (35.2%), or very severe (15.5%). Accompanying symptoms were most commonly fatigue (44.8%), chills (36.1%), exhaustion (34.9%), and fever (30.4%). Conclusion Headaches attributed to COVID-19 vaccination with the ChAdOx1 nCoV-19 (AZD1222) vaccine demonstrate an extensive and characteristic complex of symptoms. The findings have several important clinical implications for the differentiation of post-vaccinal headache and other primary as well as secondary headaches.

ZusammenfassungDie transfusionsassoziierte Graft-versus-Host-Erkrankung (ta-GvHD) ist eine seltene, aber lebensbedrohliche Nebenwirkung der Transfusion zellulärer Blutprodukte, die durch restliche Lymphozyten des Blutspenders hervorgerufen wird. Risikofaktoren für eine ta-GvHD sind eine geschwächte T-zelluläre Abwehr des Patienten, eine hohe Zahl residueller Lymphozyten im Blutprodukt und eine einseitige HLA-Inkompatibilität zwischen Spender und Empfänger. Durch eine Bestrahlung zellulärer Blutkomponenten mit mindestens 25 Gray lässt sich eine ta-GvHD sicher verhindern. In der gerade erschienenen 5. Auflage der Querschnitts-Leitlinien der Bundesärztekammer zur Therapie mit Blutkomponenten und Plasmaderivaten wurden einige Indikationen zur Bestrahlung gelockert, da Erfahrungen aus Großbritannien zeigen, dass hier auch nach Transfusion unbestrahlter Blutprodukte kein Risiko für eine ta-GvHD besteht. So stellen lymphatische Neoplasien nicht mehr generell eine Bestrahlungsindikation dar, sondern nur, wenn ein Hodgkin-Lymphom oder ein schwerer T-Zell-Defekt vorliegt oder wenn bestimmte Therapien durchgeführt werden bzw. wurden (z. B. Gabe von Purinanaloga). Zu anderen Indikationen finden sich in den revidierten Querschnitts-Leitlinien erstmals Empfehlungen zur Verwendung bestrahlter Blutkomponenten: Dies betrifft z. B. hämatoonkologische Patienten unter Therapie mit ATG oder Alemtuzumab. Der Artikel fasst den aktuellen Erkenntnisstand zur ta-GvHD kurz zusammen und erläutert die Änderungen der revidierten Querschnitts-Leitlinien.

BackgroundFatigue has an adaptive function and serves as a temporary signal to rest and save energy often in response to immune activation. It may, however, also persist in a pathological condition incurring significant burden. While subjective symptoms and scientific consensus indicate that both physical and mental determinants of motivated behavior are affected in fatigue, the underlying processes are rarely examined using objective, task-based indicators.MethodsIn three consecutive studies, including validation (N = 48) and reliability assessments (N = 27), we use an experimental task to jointly objectify reward learning and effort execution as two determinants of behavioral motivation. In addition, we tested how fatigue and its acute perturbation in response to immune activation after SARS-CoV-2 vaccination are linked to these task-based indicators of motivation in a longitudinal cross-over design (N = 55).ResultsThe validation study showed the utility of the experimental task for simultaneously assessing learning, effort exertion, and its regulation based on subjective confidence. The reliability assessment over a one-week period indicated that symptoms of fatigue and task behavior are highly reliable and that repetition effects have little impact on motivated behavior. Finally, in the vaccination trial, we found significant links between fatigue and task behavior. Baseline levels of fatigue predicted how effort is gauged in dependence of current confidence about reward outcomes, and state perturbations of fatigue in the context of the SARS-CoV-2 vaccination reduced confidence during learning. Importantly, task success was significantly lower in subjects who reported high fatigue at baseline and who additionally experienced stronger increase in fatigue in response to vaccination.DiscussionOur results demonstrate that the experimental task allows to jointly assess determinants of motivated behavior, and to link its constituent processes to subjective fatigue. This suggests that our understanding of fatigue and its perturbation due to acute immune activation can benefit from objective, task-based indicators of the underlying motivational mechanisms. Future studies could build on these findings to further deepen the understanding of neurobehavioral mechanisms underlying fatigue in the context of immune activation.

With the introduction of the virtual allocation crossmatch in the Eurotransplant (ET) region in 2023, the determination of unacceptable antigen mismatches (UAM) in kidney transplant recipients is of utmost importance for histocompatibility laboratories and transplant centers. Therefore, a joined working group of members from the German Society for Immunogenetics (Deutsche Gesellschaft für Immungenetik, DGI) and the German Transplantation Society (Deutsche Transplantationsgesellschaft, DTG) revised and updated the previous recommendations from 2015 in light of recently published evidence. Like in the previous version, a wide range of topics is covered from technical issues to clinical risk factors. This review summarizes the evidence about the prognostic value of contemporary methods for HLA antibody detection and identification, as well as the impact of UAM on waiting time, on which these recommendations are based. As no clear criteria could be determined to differentiate potentially harmful from harmless HLA antibodies, the general recommendation is to assign all HLA against which plausible antibodies are found as UAM. There is, however, a need for individualized solutions for highly immunized patients. These revised recommendations provide a list of aspects that need to be considered when assigning UAM to enable a fair and comprehensible procedure and to harmonize risk stratification prior to kidney transplantation between transplant centers.

BackgroundHeterologous vaccinations against SARS-CoV-2 with ChAdOx1 nCoV-19 and a second dose of an mRNA-based vaccine have been shown to be more immunogenic than homologous ChAdOx1 nCoV-19. In the current study, we examined the kinetics of the antibody response to the second dose of three different vaccination regimens (homologous ChAdOx1 nCoV-19 vs. ChAdOx1 nCoV-19 + BNT162b2 or mRNA-1273) against SARS-CoV-2 in a longitudinal manner; whether there are differences in latency or amplitude of the early response and which markers are most suitable to detect these responses.MethodsWe performed assays for anti-S1 IgG and IgA, anti-NCP IgG and a surrogate neutralization assay on serum samples collected from 57 participants on the day of the second vaccination as well as the following seven days.ResultsAll examined vaccination regimens induced detectable antibody responses within the examined time frame. Both heterologous regimens induced responses earlier and with a higher amplitude than homologous ChAdOx1 nCoV-19. Between the heterologous regimens, amplitudes were somewhat higher for ChAdOx1 nCoV-19 + mRNA-1273. There was no difference in latency between the IgG and IgA responses. Increases in the surrogate neutralization assay were the first changes to be detectable for all regimens and the only significant change seen for homologous ChAdOx1 nCoV-19.DiscussionBoth examined heterologous vaccination regimens are superior in immunogenicity, including the latency of the response, to homologous ChAdOx1 nCoV-19. While the IgA response has a shorter latency than the IgG response after the first dose, no such difference was found after the second dose, implying that both responses are driven by separate plasma cell populations. Early and steep increases in surrogate neutralization levels suggest that this might be a more sensitive marker for antibody responses after vaccination against SARS-CoV-2 than absolute levels of anti-S1 IgG.

The novel coronavirus SARS-CoV-2 causes the infectious disease Covid-19. Newly developed mRNA vaccines can prevent the spread of the virus. Headache is the most common neurological symptom in over 50% of those vaccinated. Detailed information about the clinical characteristics of this form of headache has not yet been described. The aim of the study is to examine in detail the clinical characteristics of headaches occurring after vaccination against Covid-19 with the BNT162b2 mRNA Covid-19 vaccine for the first time. In a multicenter observational cohort study data on the clinical features and corresponding variables were recorded using a standardized online questionnaire. The questionnaire was circulated to 12,000 residential care homes of the elderly as well as tertiary university hospitals in Germany and the United Arab Emirates. The primary outcomes of this study are the clinical features of headache after vaccination. Comorbidities, treatment with medication and sociodemographic variables are also analyzed. A total of 2349 participants reported headaches after vaccination with the BNT162b2 mRNA Covid-19 vaccine. Headaches occur an average of 18.0 ± 27.0 hours after vaccination and last an average duration of 14.2 ± 21.3 hours. Only 9.7% of those affected also report headaches resulting from previous vaccinations. In 66.6% of the participants headache occurs as a single episode. A bilateral location is indicated by 73.1% of the participants. This is most often found on the forehead (38.0%) and temples (32.1%). A pressing pain character is indicated by 49.2% and 40.7% report a dull pain character. The pain intensity is most often moderate (46.2%), severe (32.1%) or very severe (8.2%). The most common accompanying symptoms are fatigue (38.8%), exhaustion (25.7%) and muscle pain (23.4%). Headaches after Covid-19 vaccination show an extensive complex of symptoms. The constellation of accompanying symptoms together with the temporal and spatial headache characteristics delimit a distinctive headache phenotype., Graphical Abstract Graphical Abstract

Background Patients awaiting kidney transplantation are regularly screened for HLA-antibodies, but there is scarce data about the optimal interval. Methods Results from Complement-dependent cytotoxicity testing (CDC) for waitlisted patients were reviewed for increases in panel reactive antibodies (PRA) by at least 10%-points. Clinical records were screened for historic immunizing events and possible trigger factors preceding the PRA-increase. Additionally, non-pretransplanted men tested negative for HLA antibodies by solid-phase assays (SPA) out of their first two samples on the waiting list (“non-immunized men”) were evaluated for detection of HLA antibodies by SPA during their further stay on the waiting list. Results 15,360 samples from 1928 patients tested by CDC were analyzed for changes in PRA. PRA-increases occurred most frequently in patients waitlisted recently for retransplantation (annual incidence 6%). Removal of previous transplants, severe infections and/or reduced immunosuppression triggered 65% of PRA-increases during the first year after waitlisting. Transfusions accounted for 55% of PRA-increases in later years. Leucocyte-reduced red blood cell units not only boosted historic antibodies, but even induced primary immunization. In the second part of the study, 6780 samples tested by SPA from 703 non-immunized men were evaluated for development of HLA-antibodies. Only 9 men (1.3%) turned HLA antibody-positive (annual incidence 0.4%). Conclusion A uniform screening interval does not fit all: Frequencies should be highest in patients newly waitlisted for re-transplant and lowest in non-immunized men. Transfused patients should be monitored closely for development of HLA-antibodies even if leukoreduced products are used.

Background and Aims Posttransplant kidney survival depends on several risk factors. A careful immunogenetic matching and the absence of HLA donor specific antibodies (DSA) seem to determine the longevity of the transplant. Method Screening the presence of donor specific HLA antibodies in our posttransplant outpatients was implemented in 2010 (every 6 months in case of DSA free patients for two years, then yearly, and every 3 months in case of DSA + patients for two years, then twice a year). At the same time a treatment protocol was implemented, omitting reduction of immunosuppressive drugs in case of newly detected DSA, and most important with preventing steroid withdrawal in this case.The present single center study reports the long-term survival and kidney function from patients undergoing HLA-screening after transplantation between 2010 and 2016 with a follow-up until 2018. Using a Kaplan-Meier analysis patients without HLA antibodies (no HLA-ab), with HLA antibodies but without DSA (NDSA), and with donor-specific HLA antibodies (DSA) were compared by logrank-testing. Results A full dataset was obtained from 318 patients. The mean overall survival (patients and organ function) didn´t differ between the three groups, p=0.318: no HLA-ab 7.2 years (95%confidence interval 6.7;7.6), NDSA 6.6 (5.9;7.2), DSA 6.8 (6.1;7.5), overall 7.0 (6.6;7.3), events are given in Table1. Whereas the mean patient survival didn´t differ between the groups (p=0.715), the mean death-censored graft survival differed significantly, p=0.008, with a reduced transplant survival in the patients with HLA antibodies but without donorspecific antibodies: no HLA-ab 8.0 years (95%confidence interval 7.7;8.3), NDSA 7.0 (6.4;7.6), DSA 7.6 (7.1;8.2), overall 7.7 (7.4;8.0), numbers are given in Table1. Conclusion In conclusion, the presence of HLA antibodies was associated with a reduced transplant survival. Patients with HLA antibodies had a worse survival than patients with DSA undergoing HLA screening with a personalised immunosuppressive regimen. Immunosuppressive regimen of the groups, as well as other known risk factors of graft survival have to be further analysed. The results of these multivariate analyses have to be awaited to determine whether the risk for graft loss inferred by HLA antibodies is independent from other factors.

We reported previously on the widespread occurrence of anti-HLA alloantibodies of the IgA isotype (anti-HLA IgA) in the sera of solid-organ re-transplantation (re-tx) candidates (Arnold et al., ). Specifically focussing on kidney re-tx patients, we now extended our earlier findings by examining the impact of the presence and donor specificity of anti-HLA IgA on graft survival. We observed frequent concurrence of anti-HLA IgA and anti-HLA IgG in 27% of our multicenter collective of 694 kidney re-tx patients. This subgroup displayed significantly reduced graft survival as evidenced by the median time to first dialysis after transplantation (TTD 77 months) compared to patients carrying either anti-HLA IgG or IgA (TTD 102 and 94 months, respectively). In addition, donor specificity of anti-HLA IgA had a significant negative impact on graft survival (TTD 74 months) in our study. Taken together, our data strongly indicate that presence of anti-HLA IgA, in particular in conjunction with anti-HLA-IgG, in sera of kidney re-tx patients is associated with negative transplantation outcome.

Around 1 - 3% of all couples who try to have a child are affected by recurrent miscarriage. According to the WHO, recurrent miscarriage is defined as the occurrence of three or more consecutive miscarriages up to the 20th week of pregnancy. There are various causes of recurrent miscarriage; in many cases, the causes remain unclear, with the result that immunological factors are one of the possible causes discussed. For the mother's immune system, the embryo represents a semi-allogeneic transplant, as half of the embryo's genes are of paternal origin. In place of a conventional immune response, the embryo induces a secondary protection mechanism, which contributes to the successful implantation. When performing immunisation with partner lymphocytes, the patient receives an intradermal injection of her partner's prepared lymphocytes into the volar side of the forearm in order to induce immunomodulation with a consequently increased rate of pregnancy and live birth. A prerequisite for this procedure is that all other possible causes of sterility have been ruled out in advance. Due to the highly heterogeneous nature of the data, a significant benefit as a result of the immunisation cannot yet be clearly proven. However, there are signs that the therapy may be effective when using lymphocytes that have been extracted as short a time beforehand as possible. Overall, the treatment represents a safe, low-risk procedure. Following a detailed informative discussion with the couple regarding the chances of success and following a detailed review of the indication and contraindications, immunisation with partner lymphocytes can be discussed with the couple on a case-by-case basis - provided that all other possible causes of sterility have been ruled out in advance.Etwa 1 – 3% aller Kinderwunschpaare sind von einem habituellen Abortgeschehen betroffen. Dies ist laut WHO definiert als das Auftreten von 3 oder mehr aufeinanderfolgenden Aborten bis zur 20. SSW. Die Ursachen hierfür sind vielfältig, bleiben in einer Vielzahl der Fälle sogar unklar, sodass unter anderem immunologische Faktoren diskutiert werden können. Der Embryo stellt für das Immunsystem der Mutter ein semiallogenes Transplantat dar, da die Hälfte der Gene des Embryos paternaler Herkunft sind. Anstelle einer üblichen Immunantwort induziert der Embryo einen sekundären Schutzmechanismus, welcher zur erfolgreichen Implantation beiträgt. Bei der Immunisierung mit Partnerlymphozyten werden der Patientin aufbereitete Lymphozyten ihres Partners in die volare Seite des Unterarms intrakutan injiziert, um so eine Immunmodulation mit konsekutiv erhöhter Schwangerschafts- und Lebendgeburtenrate zu induzieren. Voraussetzung für dieses Verfahren ist, dass zuvor alle anderen infrage kommenden Sterilitätsursachen ausgeschlossen wurden. Aufgrund der äußerst heterogenen Datenlage kann ein signifikanter Nutzen durch die Immunisierung immer noch nicht eindeutig belegt werden. Es gibt jedoch Hinweise, dass die Therapie bei Verwendung möglichst frisch entnommener Lymphozyten wirksam sein könnte. Die Behandlung stellt insgesamt ein sicheres und risikoarmes Verfahren dar. Nach ausführlicher Aufklärung des Paares über die Erfolgsaussichten und genauer Überprüfung von Indikation und Kontraindikationen kann individuell mit dem Paar eine Immunisierung mit Partnerlymphozyten diskutiert werden – vorausgesetzt, zuvor wurden alle anderen infrage kommenden Sterilitätsursachen ausgeschlossen.

Objective To clarify whether reactivated cytomegalovirus (CMV) infections in critically ill patients lead to worse outcome or just identify more severely ill patients. If CMV has a pathogenic role, latently infected (CMV-seropositive) patients should have worse outcome than seronegative patients because only seropositive patients can experience a CMV reactivation. Design Post-hoc analysis of a prospective observational study. Setting Single university hospital. Participants The study comprised 983 consecutive patients scheduled for on-pump surgery. Interventions None. Measurements and Main Results CMV antibodies were analyzed in preoperative plasma samples. Postoperative adverse events (reintubation, low cardiac output or reinfarction, dialysis, stroke) and 30-day and 1-year mortality were evaluated prospectively. The plasma of reintubated patients and matched control patients was tested for CMV deoxyribonucleic acid, and 618 patients were found to be seropositive for CMV (63%). Among these, the risk for reintubation was increased (10% v 4%, p = 0.001). This increase remained significant after correction for confounding factors (odds ratio 2.70, p = 0.003) and was detectable from the third postoperative day throughout the whole postoperative period. Other outcome parameters were not different. Reintubated seropositive patients were more frequently CMV deoxyribonucleic acid-positive than were matched control patients (40% v 8%, p Conclusions CMV-seropositive patients had an increased risk of reintubation after cardiac surgery, which was associated with reactivations of their CMV infections. Additional studies should determine whether this complication may be prevented by monitoring of latently infected patients and administering antiviral treatment for reactivated CMV infections.

BACKGROUND DNA of human cytomegalovirus (CMV) is frequently detected in plasma of donors with primary CMV infection. It is unknown, however, whether leukoreduced blood products from these donors contain sufficient amounts of infectious virus to cause transfusion-transmitted CMV infections (TT-CMV). STUDY DESIGN AND METHODS During a 14-year period, CMV DNA-positive donations were identified as part of several previously published studies. Additionally, further donors with seroconversion were tested for CMV DNA. The serostatus of patients who had received a CMV DNA-positive blood product was determined out of pretransfusion samples. Later samples were examined for development of CMV antibodies. Patients with a follow-up of less than 140 days were also tested for CMV DNA. RESULTS A total of 221 blood products from CMV DNA-positive donations were transfused to 219 recipients. Pretransfusion samples were available for 179 patients, of whom 62 (34.6%) were seronegative. For 39 seronegative recipients of 40 blood products follow-up samples drawn at least 30 days after transfusion were available. The median duration of follow-up was 287 days (range, 38-3784 days). Thirty-six patients were still CMV seronegative in their last sample. Three patients were CMV seropositive due to passive antibody transfer by plasma rich products from seropositive donors, but CMV DNA negative in all tested samples. CONCLUSION TT-CMV was excluded in all recipients of 40 blood products from CMV DNA-positive donations. This corresponds to a 95% interval of confidence for the risk of TT-CMV of less than 7.4%. Because no patient belonged to a typical at-risk population, the results are only valid for immunocompetent subjects.

Soon, a virtual crossmatch shall replace the complement-dependent cytotoxicity (CDC) allocation crossmatch in the Eurotransplant region. To prevent positive CDC-crossmatches in the recipient centre, careful definition of unacceptable antigens is necessary. For highly sensitized patients, this is difficult by CDC alone. Assignment of all antibodies detected by sensitive assays, however, could prevent organ allocation. To assess the usefulness of the Luminex C1q-assay to prevent positive CDC-crossmatches, all CDC-crossmatches performed prior to deceased kidney transplantation in a 16-month-period were reviewed. Sera causing positive crossmatches were investigated by the C1q-assay. 31 out of 1432 crossmatches (2.2%) were positive. Sera involved in 26 positive crossmatches were available. C1q-binding donor-specific antibodies were detected in 19 sera (73.1%). The other sera were from recipients without any HLA antibodies detectable by CDC or common solid phase assays. Three patients had known Non-HLA antibodies causing positive CDC-results. Four crossmatches were only weak positive. Therefore, avoidance of donors with HLA antigens against whom C1q-binding antibodies were detected would have prevented all positive crossmatches due to HLA antibodies. Provided that all HLA specificities against which antibodies are detected by the Luminex C1q-assay are considered as unacceptable antigens, CDC-crossmatches prior to transplantation might safely be omitted in many patients. They should be maintained in highly immunized patients, however, for whom assignment of all C1q-positive antibodies as unacceptable antigens could lead to a significant delay or even prevention of transplantation.

BACKGROUND AND OBJECTIVES: The prognostic value of preformed donor-specific HLA antibodies (DSA), which are only detectable by sensitive methods, remains controversial for kidney transplantation. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The outcome of 4233 consecutive kidney transplants performed between 2012 and 2015 in 18 German transplant centers was evaluated. Most centers used a stepwise pretransplant antibody screening with bead array tests and differentiation of positive samples by single antigen assays. Using these screening results, DSA against HLA-A, -B, -C, -DRB1 and -DQB1 were determined. Data on clinical outcome and possible covariates were collected retrospectively. RESULTS: Pretransplant DSA were associated with lower overall graft survival, with a hazard ratio of 2.53 for living donation (95% confidence interval [95% CI], 1.49 to 4.29; P

SUMMARY Objective and Background To assess the performance characteristics of two fully automated human cytomegalovirus (HCMV) antibody tests. Materials and Methods Samples from negatively or not pre-screened blood donors were tested by the Biotest anti-HCMV recombinant IgG enzyme-linked immunosorbent assay (ELISA) in comparison to the Abbott Architect CMV IgG assay [chemiluminescent microparticle immunoassay (CMIA)]. For clarification, samples with discordant results between both assays were subjected to supplemental testing for anti-HCMV IgG, IgM and HCMV DNA in plasma. Results From 4938 samples tested, 362 delivered positive results in both assays (7·3%). 91 (1·8%) samples were discordant. Of 43 (two not further tested) samples positive only by ELISA, 41 were false positive, one true positive and one indeterminate. Of 45 (one not further tested) samples positive only by CMIA, 20 were false positive, 9 indeterminate and 16 true positive. Anti-HCMV IgM and HCMV DNA testing from the plasma were negative in indeterminate samples. Considering the results of supplemental testing, the CMIA achieved altogether better results concerning resolved sensitivity, resolved specificity as well as negative predictive value. Both assays had an inferior positive predictive value, with a better result for CMIA. Conclusion Overall, the performance characteristics of the CMIA were better than those of the ELISA. Owing to the inferior positive predictive value, positive test results require confirmation if blood products from donors with remote HCMV infection should be administered.

Background Donors with short interdonation intervals (e.g., apheresis donors) have an increased risk of window period donations. The frequency of cytomegalovirus (CMV) window period donations is important information to decide whether selection of seronegative donors might be advantageous for patients at risk for transfusion-transmitted CMV infections (TT-CMV). Study Design and Methods CMV seroconversion in 93 donors with positive results in routine CMV antibody testing within at most 35 days after the last seronegative sample was evaluated by Western blot and/or a second antibody test. In donors with unconfirmed seroconversion, an additional later sample was tested. Concentration of CMV DNA was determined in pre- and postseroconversion samples. Results CMV seroconversion was confirmed in 12 donors (13%). Among these, the last seronegative sample was CMV DNA positive in three donors (25%, below 30 IU/mL). The first seropositive sample was CMV DNA positive in 10 donors (83%, maximum 1600 IU/mL). Both prevalence and median concentration of CMV DNA were higher in the first seropositive sample (p = 0.004 and p = 0.02), with maximum concentrations being reached about 2 weeks after seroconversion. No CMV DNA was detected in samples from donors with unconfirmed seroconversion. Conclusion At least in donors with short interdonation intervals, most suspected CMV seroconversions are due to false-positive results of the screening test. As window period donations are rare and contain less CMV DNA than the first seropositive donation, avoidance of blood products from primarily seropositive donors is especially helpful to avoid TT-CMV if donors with short interdonation intervals are concerned.

Background. The assignment of human leucocyte antigens (HLAs) against which antibodies are detected as unacceptable antigens (UAGs) avoids allocation of HLA-incompatible allografts. There is uncertainty as to what extent UAGs decrease the probability of receiving a kidney offer. Methods. Kidney transplantations in 3264 patients on the waiting lists of six German transplant centres were evaluated for a period of at least 2 years. The proportion of excluded offers due to UAGs was calculated as virtual panel-reactive antibodies (vPRAs). Results. In the common Eurotransplant Kidney Allocation Scheme, the transplant probability was unaffected by vPRAs in exploratory univariate analyses. In the multivariable model, a 1% increase in vPRA values was outweighed by an additional waiting time of 2.5 weeks. The model was confirmed using an external validation cohort of 1521 patients from seven centres. If only patients with standard risk were considered (e.g. no simultaneous transplantation of other organs), only 1.3 weeks additional waiting time was needed. In the Eurotransplant Senior Program, patients with vPRA values >50% had a strongly reduced transplant probability in the unadjusted analyses. In the multivariable model, a 1% increase in vPRA values was outweighed by an additional waiting time of 5 weeks. Conclusions: This study demonstrates that the assignment of UAGs decreases the transplant probability in both main Eurotransplant allocation programs because of insufficient compensatory mechanisms. At present, for immunized patients, a prolonged waiting time has to be weighed against the increased immunologic risk due to donor-specific antibodies not assigned as UAGs.

BACKGROUND: Capillary hemoglobin (Hb) measurement before admission for whole blood donation is performed in many blood donation services, in spite of several studies reporting many donors with low Hb values being missed by capillary Hb screening. STUDY DESIGN AND METHODS: Predonation capillary and venous Hb levels of 9209 first-time donors presenting for whole blood donation were compared. Hb testing was conducted by photometric determination of finger-stick samples with a photometric method (Donor Checker, HemoCue) and using a hematology analyzer for venous samples. RESULTS: Both capillary and venous Hb measurements were available for 8910 donors (96.8%). The arithmetic mean deviation of the capillary Hb from the venous sample was +1.5 ± 6.8 g/L, and the mean deviation was 5.4 ± 4.5 g/L. In 7865 donors (88%), capillary and venous Hb values differed less than 10 g/L from each other, but in 86 donors (1.0%) the difference was at least 20 g/L, and in 10 donors (0.1%), even 30 g/L or more. In 93.3% of females and 98.7% of males, the categorization as having sufficient or too low Hb for blood donation was concordant between capillary and venous measurements. A total of 34.4% of donors with too low results of capillary Hb screening had sufficient venous Hb levels. Only one donor (0.01%) passed the capillary Hb screening despite venous Hb values below 110 g/L. CONCLUSION: The currently available methods for capillary Hb screening allow reliable determination of predonation Hb values under routine conditions. Additional venous Hb measurements in donors with too low capillary Hb values might reduce the rate of deferred donors by approximately one-third.

BACKGROUND: Screening of blood donations for antibodies against hepatitis B core antigen (anti-HBc) is an accepted method to prevent some transfusion-transmitted hepatitis B virus (HBV) infections. However, anti-HBc testing may result in donor loss due to unspecific results in the currently available anti-HBc tests. Algorithms to distinguish true-positive from false-positive results and for reentry of those donors who tested false anti-HBc positive were evaluated retrospectively. STUDY DESIGN AND METHODS: Samples that tested reactive for anti-HBc by chemiluminescent microparticle immunoassay (CMIA) were investigated for anti-HBc by microparticle immunoassay, for anti-HBs and hepatitis B surface antigen (HBsAg) by CMIA, and for HBV DNA by individual-donor nucleic acid testing. Results were classified true positive, indeterminate, and false positive for anti-HBc. Donors who tested indeterminate and false positive were admitted for reentry if follow-up testing for anti-HBc became negative and no further evidence for an HBV infection was apparent. RESULTS: A total of 554 of 148,000 samples, taken from 30,000 individuals within 3 years tested reactive for anti-HBc by CMIA. Of those, 553 could be further classified: 142 (26%) true positive, 76 (14%) indeterminate, and 335 (60%) false positive. A total of 214 of 411 (52%) samples termed indeterminate or false positive were admitted for reentry and able to provide further donations. In one donor, anti-HBc–positive/HBsAg- and HBV DNA–negative HBV DNA was detectable during follow-up. CONCLUSION: According to our proposed algorithm, 26% of anti-HBc–reactive results tested by CMIA were true positive. Many donors tested indeterminate or false positive can provide future donations if our proposed algorithm for reentry is applied. One donor at risk for transmitting HBV was identified solely by anti-HBc testing.

Background and Objectives As cytomegalovirus (CMV) DNA is frequently detectable in the plasma of recently infected sero-positive blood donors, information concerning primary CMV infection is important for the identification of possibly infectious donors. Materials and Methods Monitoring of 17 982 donors for CMV antibodies and DNA in plasma identified 14 subjects with ongoing primary CMV infection. Thirteen donors were interrogated for possible sources of infection and CMV-related symptoms, and monitored for CMV antigens, CMV DNA in plasma, leucocytes and urine, course of IgG and IgM antibodies as well as markers of systemic infection and parameters of organ function. Results CMV antigens and DNA were detectable in peripheral blood for up to 54 and 269 days respectively. Clearance of CMV DNA from blood correlated with clearance of IgM antibodies, development of IgG antibodies against the membrane glycoprotein gB and development of high avidity IgG antibodies. Eighty-five percent of subjects with primary CMV infection, but even 69% of matched controls reported possibly CMV-related symptoms. Sixty-two and 23%, respectively, had contact with possible sources of infection. One donor developed a febrile illness accompanied by increased levels of CMV DNA in peripheral blood 2 to 3 weeks after seroconversion. In other donors, neither markers of systemic infection nor parameters of organ function correlated with the course of CMV DNA and antigens. Conclusion Potentially infectious donors can be identified by measuring CMV DNA, IgM antibodies or avidity of IgG antibodies. Alternatively, blood products donated during the first year after seroconversion should not be used for immunocompromised patients.

This work describes an optimization of a simple photometric determination of iodine concentrations in urine using a modified ceric arsenite method with ammonium persulfate as oxidant. By means of this sensitive method iodine concentrations can be determined in very small specimens (50 microL). Urine samples (105) collected from a mixed population, were analyzed for urine iodine content by the optimized ammonium persulfate method, a Technicon Autoanalyzer II and a paired-ion-RP HPLC. We found that the precision of this optimized ammonium persulfate method yields inter assay CVs of10% for urinary iodine concentrations10 microg/dL. Recovery of [123I]iodide added to urine in vitro was 100.9 +/- 2.4%. The detection limit was 0.0029 microg iodine. There was a high correlation between all three methods (r0.94 in any case) and the interpretation of the results was consistent. We conclude that this simple, manual ammonium persulfate method is suitable for urinary iodine analysis and can be performed in any routine clinical laboratory.

Positive end-expiratory pressure (PEEP) during mechanical ventilation may impose different degrees of stress on healthy lungs. On the assumption that stress is reflected by cytokine production, we performed a translational study investigating the effect of PEEP on bronchoalveolar and systemic mediator levels in isolated perfused mouse lungs (IPL) and in patients with healthy lungs.(Part I) IPL were ventilated with end-expiratory pressures of 0, 3, 6, or 10 cm H2O and end-inspiratory pressure (EIP) levels of 10 or 25 cm H2O. Interleukin (IL)-6 and macrophage inflammatory protein-2 concentrations in the venous effluate were monitored. (Part II) Patients (nonsmokers) scheduled for elective otorhinolaryngology surgery (duration90 min) were randomized to receive either ventilation with zero end-expiratory pressure or PEEP (10 cm H2O). Mediators in bronchoalveolar lavage, nuclear factor kappaB, (NF-kappaB)-activation in alveolar macrophages and circulating systemic mediators were monitored. Control patients underwent bronchoalveolar lavage after intubation.In the IPL, mediator concentrations increased with increasing end-expiratory pressure at an EIP of 10 cm H2O, but decreased at 25 cm H2O EIP. In patients, bronchoalveolar IL-6, monocyte chemoattractant protein-1, and granulocyte monocyte-colony stimulating factor were increased by ventilation regardless of the PEEP level. IL-6 and IL-8 levels were moderately increased by PEEP but not zero end-expiratory pressure. Nuclear factor kappaB DNA binding activity in alveolar macrophages and systemic mediator levels did not change.On the basis of the premise that cytokine levels may indicate mechanical stress, our findings indicate that even low tidal volume ventilation causes some stress. PEEP is beneficial at high inspiratory pressure, but imposes moderate stress at low inspiratory pressure.

Traditionally, leukoreduction and selection of blood products from seronegative donors have been used as alternative strategies to reduce the risk of transfusion-transmitted cytomegalovirus infections (TT-CMV) in atrisk patients. After the introduction of universal leukoreduction for red blood cell and platelet concentrates in Germany, a controversy evolved as to whether the additional selection of blood products from seronegative donors would reduce or even increase the risk of TT-CMV. This review summarizes the current knowledge about CMV infections in blood donors and the implications of this information on the effect of potential transfusion strategies. Even though there are conflicting data about the incidence of TT-CMV remaining after the introduction of leukodepletion, it has been clearly shown that both prevalence and concentration of CMV DNA in peripheral blood are highest in newly seropositive donors. Therefore, avoidance of blood products from these donors is the most important goal of any transfusion strategy. This goal can be reached by: i) selection of blood products from seronegative donors, ii) provision of CMV DNA-negative blood products, or iii) provision of blood from long-term seropositive donors. In cases of suspected TT-CMV, all implicated donors should be investigated carefully to gather further knowledge on which donors confer the lowest risk for TT-CMV.

Background Cytomegalovirus (CMV) DNA is frequently detected in plasma of newly seropositive donors. Selection of leukoreduced blood products from donors with remote CMV infection could avoid transfusion-transmitted CMV infections (TT-CMV) due to primarily infected donors. However, there are no data about the prevalence of reactivations in long-term seropositive donors compared to the incidence of window period donations in seronegative donors. Therefore, the optimal transfusion strategy for at-risk patients is unclear. Study Design and Methods Whole blood samples from 22,904 donations were tested for CMV DNA, and CMV DNA–positive donations were categorized as donations from 1) seronegative donors, 2) newly seropositive donors, and 3) long-term seropositive donors. Results Twenty-one donors were reproducibly CMV DNA–positive (0.09%). Frequency of detection and concentration of CMV DNA in whole blood were comparable for seronegative and long-term seropositive donors. Nonreproducibly positive results for CMV DNA in whole blood were more frequent in long-term seropositive donors (0.16% vs. 0.01%, p

Cytomegalovirus (CMV) DNA is frequently detected in plasma of newly seropositive donors. Selection of leukoreduced blood products from donors with remote CMV infection could avoid transfusion-transmitted CMV infections (TT-CMV) due to primarily infected donors. However, there are no data about the prevalence of reactivations in long-term seropositive donors compared to the incidence of window period donations in seronegative donors. Therefore, the optimal transfusion strategy for at-risk patients is unclear.Whole blood samples from 22,904 donations were tested for CMV DNA, and CMV DNA-positive donations were categorized as donations from 1) seronegative donors, 2) newly seropositive donors, and 3) long-term seropositive donors.Twenty-one donors were reproducibly CMV DNA-positive (0.09%). Frequency of detection and concentration of CMV DNA in whole blood were comparable for seronegative and long-term seropositive donors. Nonreproducibly positive results for CMV DNA in whole blood were more frequent in long-term seropositive donors (0.16% vs. 0.01%, p0.01). Only low concentrations of CMV DNA in plasma were detectable in two seronegative donors and one long-term seropositive donor. Highest concentrations of CMV DNA in both whole blood and plasma, however, were found in newly seropositive donors.Prevalences of window period donations among seronegative donors and reactivations among long-term seropositive donors, as well as the CMV DNA concentration in whole blood and plasma samples from these donors, are comparable. Therefore, blood products from both groups could be used for patients at risk for TT-CMV, while those of newly seropositive donors seem to bear an increased risk.

Summary In this multicentre study, sera from 803 retransplant candidates, including 775 kidney transplant recipients, were analysed with regard to the presence and specificity of anti-HLA alloantibodies of the IgA isotype using a modified microsphere-based platform. Of the kidney recipients, nearly one-third (n = 237, 31%) had IgA alloantibodies. Mostly, these antibodies were found in sera that also harboured IgG alloantibodies that could be found in a total of 572 (74%) of patients. Interestingly, IgA anti-HLA antibodies were preferentially targeting HLA class I antigens in contrast to those of the IgG isotype, which targeted mostly both HLA class I and II antigens. Donor specificity of the IgA alloantibodies could be established for over half of the 237 patients with IgA alloantibodies (n = 124, 52%). A further 58 patients had specificities against HLA-C or HLA-DP, for which no information regarding donor typing was available. In summary, these data showed in a large cohort of retransplant candidates that IgA alloantibodies occur in about one-third of patients, about half of these antibodies being donor specific.

Donors with short interdonation intervals (e.g., apheresis donors) have an increased risk of window period donations. The frequency of cytomegalovirus (CMV) window period donations is important information to decide whether selection of seronegative donors might be advantageous for patients at risk for transfusion-transmitted CMV infections (TT-CMV).CMV seroconversion in 93 donors with positive results in routine CMV antibody testing within at most 35 days after the last seronegative sample was evaluated by Western blot and/or a second antibody test. In donors with unconfirmed seroconversion, an additional later sample was tested. Concentration of CMV DNA was determined in pre- and postseroconversion samples.CMV seroconversion was confirmed in 12 donors (13%). Among these, the last seronegative sample was CMV DNA positive in three donors (25%, below 30 IU/mL). The first seropositive sample was CMV DNA positive in 10 donors (83%, maximum 1600 IU/mL). Both prevalence and median concentration of CMV DNA were higher in the first seropositive sample (p = 0.004 and p = 0.02), with maximum concentrations being reached about 2 weeks after seroconversion. No CMV DNA was detected in samples from donors with unconfirmed seroconversion.At least in donors with short interdonation intervals, most suspected CMV seroconversions are due to false-positive results of the screening test. As window period donations are rare and contain less CMV DNA than the first seropositive donation, avoidance of blood products from primarily seropositive donors is especially helpful to avoid TT-CMV if donors with short interdonation intervals are concerned.

HLA antibody screening is conducted routinely prior to kidney transplantation, but the comparative prognostic value and cost-effectiveness of different methods are unclear. Pre-transplant sera of 141 patients transplanted between 1998 and 2000 were screened by ELISA and Luminex assays, and antibody specificities of reactive sera determined using bead array techniques. ELISA screening detected donor-specific antibodies (DSA) in 19 patients, who had a higher incidence of impaired graft function (60% vs. 20%, p = 0.04) and antibody-mediated rejection (AMR) within 90 d after transplantation (AMR, 35% vs. 5%, p = 0.02). Luminex screening detected eight additional patients with DSA, among those one with AMR. Six of eight patients with Luminex-only-DSA reported no prior immunizing events. Death-censored graft survival was shorter only in patients with DSA and AMR (median, 1.7 yr instead of between 9.5 and 11.0 yr for patients without DSA or patients with DSA but no AMR, p < 0.001). Material costs per detected clinically relevant DSA were about 57% higher for Luminex screening, but this increase could be avoided by modifying the cut-off recommended by the manufacturer. Conclusively, specification of antibodies only in sera reactive in screening tests was cost-effective to prevent shortened graft survival. Preformed DSA were only harmful if AMR was diagnosed within 90 d after transplantation.

Capillary hemoglobin (Hb) measurement before admission for whole blood donation is performed in many blood donation services, in spite of several studies reporting many donors with low Hb values being missed by capillary Hb screening.Predonation capillary and venous Hb levels of 9209 first-time donors presenting for whole blood donation were compared. Hb testing was conducted by photometric determination of finger-stick samples with a photometric method (Donor Checker, HemoCue) and using a hematology analyzer for venous samples.Both capillary and venous Hb measurements were available for 8910 donors (96.8%). The arithmetic mean deviation of the capillary Hb from the venous sample was +1.5 ± 6.8 g/L, and the mean deviation was 5.4 ± 4.5 g/L. In 7865 donors (88%), capillary and venous Hb values differed less than 10 g/L from each other, but in 86 donors (1.0%) the difference was at least 20 g/L, and in 10 donors (0.1%), even 30 g/L or more. In 93.3% of females and 98.7% of males, the categorization as having sufficient or too low Hb for blood donation was concordant between capillary and venous measurements. A total of 34.4% of donors with too low results of capillary Hb screening had sufficient venous Hb levels. Only one donor (0.01%) passed the capillary Hb screening despite venous Hb values below 110 g/L.The currently available methods for capillary Hb screening allow reliable determination of predonation Hb values under routine conditions. Additional venous Hb measurements in donors with too low capillary Hb values might reduce the rate of deferred donors by approximately one-third.

Screening of blood donations for antibodies against hepatitis B core antigen (anti-HBc) is an accepted method to prevent some transfusion-transmitted hepatitis B virus (HBV) infections. However, anti-HBc testing may result in donor loss due to unspecific results in the currently available anti-HBc tests. Algorithms to distinguish true-positive from false-positive results and for reentry of those donors who tested false anti-HBc positive were evaluated retrospectively.Samples that tested reactive for anti-HBc by chemiluminescent microparticle immunoassay (CMIA) were investigated for anti-HBc by microparticle immunoassay, for anti-HBs and hepatitis B surface antigen (HBsAg) by CMIA, and for HBV DNA by individual-donor nucleic acid testing. Results were classified true positive, indeterminate, and false positive for anti-HBc. Donors who tested indeterminate and false positive were admitted for reentry if follow-up testing for anti-HBc became negative and no further evidence for an HBV infection was apparent.A total of 554 of 148,000 samples, taken from 30,000 individuals within 3 years tested reactive for anti-HBc by CMIA. Of those, 553 could be further classified: 142 (26%) true positive, 76 (14%) indeterminate, and 335 (60%) false positive. A total of 214 of 411 (52%) samples termed indeterminate or false positive were admitted for reentry and able to provide further donations. In one donor, anti-HBc-positive/HBsAg- and HBV DNA-negative HBV DNA was detectable during follow-up.According to our proposed algorithm, 26% of anti-HBc-reactive results tested by CMIA were true positive. Many donors tested indeterminate or false positive can provide future donations if our proposed algorithm for reentry is applied. One donor at risk for transmitting HBV was identified solely by anti-HBc testing.

Objective: To determine the prevalence and impact on patient outcome of active human cytomegalovirus infections in patients with prolonged treatment in an intensive care unit. Design: Retrospective analysis of stored plasma samples. Setting: Anesthesiological intensive care unit of a university hospital. Patients: All 138 patients treated for at least 14 days (of a total of 4940 patients admitted during the study period). Immunocompromised patients and patients with inconclusive results for cytomegalovirus DNA were excluded. Interventions: None. Measurements and Main Results: Stored plasma samples of patients with prolonged intensive care unit stay were tested for cytomegalovirus DNA. Sixty-four of 255 evaluable samples from 99 immunocompetent patients tested cytomegalovirus DNA-positive with a mean DNA concentration of 8,600 genome equivalents per milliliter. Active cytomegalovirus infection was diagnosed by reproducibly positive results in 35 patients (35%). Only one case had been diagnosed clinically. Patients with and without active cytomegalovirus infection were not significantly different in parameters, such as age, sex, admission category, source of admission, or comorbidities. Even review of specific surgical procedures or the use of a heart-lung–machine showed no significant differences between the groups. The mortality rate in patients with cytomegalovirus infection was significantly increased (28.6% vs. 10.9%, p = 0.048), and surviving patients had a longer intensive care unit stay (32.6 vs. 22.1 days, p Conclusions: Active cytomegalovirus infection is a frequent but seldom diagnosed finding in surgical patients with prolonged intensive care unit stay, which is associated with increased mortality and prolonged intensive care unit stay of surviving patients.

BACKGROUND: Human cytomegalovirus (CMV) is considered to latently infect blood cells. Transfusion-transmitted infection (TT-CMV) of immunocompromised patients occurs despite the use of CMV-seronegative or leukoreduced units. STUDY DESIGN AND METHODS: The prevalence of CMV DNA in plasma was investigated in 82 blood donors who had previously been seronegative for CMV and showed anti-CMV immunoglobulin G for the first time, 598 blood donors who were seropositive for at least 1 year, and 150 seronegative blood donors. In a second part of the study, the overall prevalence of CMV DNA in blood donations was assessed based on 31,745 donations. RESULTS: CMV DNA was repeatedly detected in plasma samples of 44 percent of newly seropositive donors (12%-62%, depending on the interval to the last seronegative donation). All steadily seropositive or seronegative donors were negative for the presence of CMV DNA. Detection of CMV DNA in connection with seroconversion was accompanied by significantly increased neopterin, increased alanine aminotransferase, and reduced white blood cell counts, but the sensitivity of these surrogate markers was only 71 percent. The overall prevalence of CMV DNA in blood products due to primary CMV infection of donors was at least 0.13 percent. CONCLUSION: Viremia of newly seropositive donors may be an important reason for the residual risk of TT-CMV despite leukoreduction. Furthermore, transfusion of WBC-reduced blood components from seronegative donors could imply a greater risk of TT-CMV than transfusion of WBC-reduced blood from donors who have been seropositive for at least 1 year, because window-phase donations but no reactivation could be detected in this study.

The human leukocyte antigen (HLA) complex contains the most polymorphic genes in the human genome. The classical HLA class I and II genes define the specificity of adaptive immune responses. Genetic variation at the HLA genes is associated with susceptibility to autoimmune and infectious diseases and plays a major role in transplantation medicine and immunology. Currently, the HLA genes are characterized using Sanger- or next-generation sequencing (NGS) of a limited amplicon repertoire or labeled oligonucleotides for allele-specific sequences. High-quality NGS-based methods are in proprietary use and not publicly available. Here, we introduce the first highly automated open-kit/open-source HLA-typing method for NGS. The method employs in-solution targeted capturing of the classical class I (HLA-A, HLA-B, HLA-C) and class II HLA genes (HLA-DRB1, HLA-DQA1, HLA-DQB1, HLA-DPA1, HLA-DPB1). The calling algorithm allows for highly confident allele-calling to three-field resolution (cDNA nucleotide variants). The method was validated on 357 commercially available DNA samples with known HLA alleles obtained by classical typing. Our results showed on average an accurate allele call rate of 0.99 in a fully automated manner, identifying also errors in the reference data. Finally, our method provides the flexibility to add further enrichment target regions.