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After recovery, mild and severe COVID-19 diseases are associated with long-term effects on the host immune system, such as prolonged T-cell activation or accumulation of autoantibodies. In this study, we show that mild SARS-CoV-2 infections, but not SARS-CoV-2 spike mRNA vaccinations, cause durable atopic risk factors such as a systemic Th2- and Th17-type environment as well as activation of B cells responsive of IgE against aeroallergens from house dust mite and mold. At an average of 100 days post mild SARS-CoV-2 infections, anti-mold responses were associated with low IL-13 levels and increased pro-inflammatory IL-6 titers. Acutely severely ill COVID-19 patients instead showed no evidence of atopic reactions. Considering convalescents of mild COVID-19 courses and mRNA-vaccinated individuals together, IL-13 was the predominant significantly upregulated factor, likely shaping SARS-CoV-2 immunity. Application of multiple regression analysis revealed that the IL-13 levels of both groups were determined by the Th17-type cytokines IL-17A and IL-22. Taken together, these results implicate a critical role for IL-13 in the aftermath of SARS-CoV-2 mild infections and mRNA vaccinations, conferring protection against airway directed, atopic side reactions that occur in mildly experienced COVID-19.

PurposeThe SeMaCo study (Serologische Untersuchungen bei Blutspendern des Großraums Magdeburg auf Antikörper gegen SARS-CoV-2), a prospective, longitudinal cohort study with four survey phases spanning 3–5 months each over a period of 22 months, extends the spectrum of seroepidemiological studies in Germany. We present here a careful characterisation of the initial survey phase of the cohort to provide baseline data on infection incidence and obtained from questionnaires, focussing in particular on the attitude towards COVID-19 vaccinations, the vaccination success and the vaccination acceptance.ParticipantsA total of 2195 individual blood donors from the donor pool of the blood donation service of the University Hospital Magdeburg were enrolled in the initial survey phase from 20 January 2021 to 30 April 2021. 2138 participants gave sociodemographic/contact data (51.7% male, mean age 44 years) and 2082 participants answered the vaccination questionnaire.Findings to dateOut of 2195 participants with antibody results, 1909 (87.0%) were antibody negative. The remaining 286 subjects (13.0%) were either antibody-positive and vaccinated (160/286; 55.9%) or antibody-positive without vaccination information (17/286; 5.9%) or antibody-positive and unvaccinated (109/286; 38.1%). The latter result reflects the rate of true or highly probable SARS-CoV-2 infections in our initial study cohort.Future plansThe study primarily aims to measure the prevalence and long-term kinetics of IgG-antibodies against SARS-CoV-2. Including the baseline, the study foresees four survey periods of 3–4 months each. At each visit, we will assess the blood donors’ attitude towards vaccination, the antibody response following vaccination and/or infection, as well as undesired vaccination effects. We aim to test the same participants during the survey periods by repeated invitations for blood donation to ensure a long-term (follow-up) in as many study participants as possible. After the four survey phases, a longitudinal data set will be created that reflects the course of the antibody levels/frequencies as well as the infection and vaccination incidence.Trial registration numberDRKS00023263.

Objectives The optimal diagnostic specimen to detect SARS-CoV-2 by PCR in the upper respiratory tract is unclear. Mouthwash fluid has been reported as an alternative to nasopharyngeal and oropharyngeal swabs. We compared mouthwash fluid with a combined oro-nasopharyngeal swab regarding test performance. Methods In a large refugee facility, we retested individuals with a previous positive test for SARS-CoV-2 and their quarantined close contacts. All individuals were asymptomatic at the time of testing. First, a mouthwash (gargling for at least 5 s) with sterile water was performed. Then, with a single flocked swab the back of the throat and subsequently the nasopharynx were sampled. Samples were inactivated and analysed on a Roche cobas 6800® system with the Roche SARS-CoV-2 test. Results Of 76 individuals, 39 (51%) tested positive for SARS-CoV-2 by oro-nasopharyngeal swab. Mouthwash detected 13 of 76 (17%) infections, but did not detect any additional infection. Samples that were positive in both tests, had lower cycle threshold (Ct)-values for oro-nasopharyngeal samples, indicating a higher virus concentration, compared to samples only positive in oro-nasopharyngeal swabs. Conclusion Mouthwash is not as sensitive as combined oro-nasopharyngeal swab in detecting upper respiratory tract infection.

Newborns are highly susceptible to infections; however, the underlying mechanisms that regulate the anti-microbial T-helper cells shortly after birth remain incompletely understood. To address neonatal antigen-specific human T-cell responses against bacteria, Staphylococcus aureus (S. aureus) was used as a model pathogen and comparatively analyzed in terms of the polyclonal staphylococcal enterotoxin B (SEB) superantigen responses. Here, we report that neonatal CD4 T-cells perform activation-induced events upon S. aureus/APC-encounter including the expression of CD40L and PD-1, as well as the production of Th1 cytokines, concomitant to T-cell proliferation. The application of a multiple regression analysis revealed that the proliferation of neonatal T-helper cells was determined by sex, IL-2 receptor expression and the impact of the PD-1/PD-L1 blockade. Indeed, the treatment of S. aureus-activated neonatal T-helper cells with PD-1 and PD-L1 blocking antibodies revealed the specific regulation of the immediate neonatal T-cell responses with respect to the proliferation and frequencies of IFNγ producers, which resembled in part the response of adults’ memory T-cells. Intriguingly, the generation of multifunctional T-helper cells was regulated by the PD-1/PD-L1 axis exclusively in the neonatal CD4 T-cell lineage. Together, albeit missing memory T-cells in neonates, their unexperienced CD4 T-cells are well adapted to mount immediate and strong anti-bacterial responses that are tightly controlled by the PD-1/PD-L1 axis, thereby resembling the regulation of recalled memory T-cells of adults.

ObjectiveGenetic determinants of fibrin clot formation and fibrinolysis have an impact on local and systemic inflammatory response. The aim of the present study was to assess whether coagulation-related genotypes affect the predictive value of C-reactive protein (CRP) in regards of radiographic spinal progression in axial spondyloarthritis (axSpA).MethodsTwo hundred and eight patients with axSpA from the German Spondyloarthritis Inception Cohort were characterised for genotypes of α-fibrinogen, β-fibrinogen (FGB) and γ-fibrinogen, factor XIII A-subunit (F13A) and α2-antiplasmin (A2AP). The relation between CRP levels and radiographic spinal progression defined as worsening of the modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS) by ≥2 points over 2 years was assessed in dependence on the respective genetic background in logistic regression analyses.ResultsOverall, CRP was associated with mSASSS progression ≥2 points: time-averaged CRP ≥10 mg/L, OR: 3.32, 95% CI 1.35 to 8.13. After stratification for coagulation-related genotypes, CRP was strongly associated with mSASSS progression in individuals predisposed to form loose, fibrinolysis-susceptible fibrin clots (FGB rs1800790GG, OR: 6.86, 95% CI 2.08 to 22.6; A2AP 6Trp, OR: 5.86, 95% CI 1.63 to 21.0; F13A 34Leu, OR: 8.72, 95% CI 1.69 to 45.1), while in genotypes predisposing to stable fibrin clots, the association was absent or weak (FGB rs1800790A, OR: 0.83, 95% CI 0.14 to 4.84; A2AP 6Arg/Arg, OR: 1.47, 95% CI 0.35 to 6.19; F13A 34Val/Val, OR: 1.72, 95% CI 0.52 to 5.71).ConclusionsElevated CRP levels seem to be clearly associated with radiographic spinal progression only if patients are predisposed for loose fibrin clots with high susceptibility to fibrinolysis.

In the placebo-controlled, double-blind BOne marrOw transfer to enhance ST-elevation infarct regeneration (BOOST) 2 trial, intracoronary autologous bone marrow cell (BMC) transfer did not improve recovery of left ventricular ejection fraction (LVEF) at 6 months in patients with ST-elevation myocardial infarction (STEMI) and moderately reduced LVEF. Regional myocardial perfusion as determined by adenosine stress perfusion cardiac magnetic resonance imaging (S-CMR) may be more sensitive than global LVEF in detecting BMC treatment effects. Here, we sought to evaluate (i) the changes of myocardial perfusion in the infarct area over time (ii) the effects of BMC therapy on infarct perfusion, and (iii) the relation of infarct perfusion to LVEF recovery at 6 months. In 51 patients from BOOST-2 (placebo, n = 10; BMC, n = 41), S-CMR was performed 5.1 ± 2.9 days after PCI (before placebo/BMC treatment) and after 6 months. Infarct perfusion improved from baseline to 6 months in the overall patient cohort as reflected by the semi-quantitative parameters, perfusion defect–infarct size ratio (change from 0.54 ± 0.20 to 0.43 ± 0.22; P = 0.006) and perfusion defect–upslope ratio (0.54 ± 0.23 to 0.68 ± 0.22; P

ZusammenfassungPräparative Hämapheresen sind in vielen transfusionsmedizinischen Einrichtungen Standardverfahren zur modernen Herstellung von Blutprodukten. Sie kommen jährlich weltweit millionenfach zum Einsatz. Spenderreaktionen im Rahmen von präparativen Hämapheresen bei gesunden unverwandten Spendern sind dank moderner Apheresetechnik und hohen Prozessstandards heute ganz überwiegend als mild und klinisch nicht signifikant einzustufen. Die Kenntnis dieser, aber auch der seltenen schwerwiegenden Reaktionen, ihrer Symptome und ihrer Behandlung ist wichtig, um Spendersicherheit und Spendercompliance zu gewährleisten und Verfahrensabbrüche zu reduzieren bzw. zu vermeiden.

ZusammenfassungAdenovirus-Infektionen (AdV-Infektionen) nach pädiatrischer hämatopoetischer Stammzelltransplantation sind häufig und treten in 10 – 20 % aller allogenen Transplantationen auf (bis 30 % der T-Zell-depletierten Transplantationen). Sie sind mit einer hohen Rate an Morbidität und Mortalität verbunden. Da die antivirale Therapie nur virostatisch wirkt und durch erhebliche Toxizität eine limitierende Behandlungsoption darstellt, ist die virusspezifische T-Zell-Therapie als sichere und gut verträgliche komplikationsarme Option sehr attraktiv. Unsere 4-jährige Patientin entwickelte 3 Wochen nach allogener hämatopoetischer Transplantation vom 10/10-HLA-passenden Registerspender eine disseminierte AdV-Infektion (Enteritis, persistierendes Fieber, Sepsis). Eine antivirale Therapie mit Cidofovir zeigte eine gute Wirksamkeit, musste aber wegen erheblicher Nephrotoxizität abgebrochen werden. Unter einer Überbrückungstherapie bis zur geplanten Gabe AdV-spezifischer T-Zellen mit Brincidofovir (BCV) stabilisierte sich die AdV-Last im Blut, blieb aber signifikant hoch. AdV-spezifische T-Zellen der HLA-haploidenten Mutter wurden nach Stimulation mit AdV-Hexon-Peptid-Gemisch über Interferon-γ-Zytokinsekretion (Interferon-γ: IFN-γ) und immunmagnetische Separation aufgereinigt und 1-malig in einem Frischpräparat mit einer Dosis von 2,5 × 104CD3+-Zellen/kg Körpergewicht verabreicht. Die Anwendung verlief komplikationslos und führte zu einer schnellen Elimination des Virus im Blut (ca. nach 2 Wochen). Nahezu zeitgleich (ca. 3 Wochen nach der AdV-spezifischen T-Zell-Gabe) konnten erstmals AdV-spezifische T-Zellen im Patientenblut nachgewiesen werden. Der adoptive Transfer AdV-spezifischer T-Zellen vom HLA-teilpassenden Familienspender als „Third-Party-Spender“ stellt eine rasch verfügbare und wenig toxische Behandlungsoption für Patienten mit refraktärer AdV-Erkrankung dar.

The variability in resolution of SARS-CoV-2-infections between individuals neither is comprehended, nor are the long-term immunological consequences. To assess the long-term impact of a SARS-CoV-2-infection on the immune system, we conducted a prospective study of 80 acute and former SARS-CoV-2 infected individuals and 39 unexposed donors to evaluate autoantibody responses and immune composition. Autoantibody levels against cyclic citrullinated peptide (CCP), a specific predictor for rheumatoid arthritis (RA), were significantly (p = 0.035) elevated in convalescents only, whereas both acute COVID-19 patients and long-term convalescents showed critically increased levels of anti-tissue transglutaminase (TG), a specific predictor of celiac disease (CD) (p = 0.002). Both, anti-CCP and anti-TG antibody levels were still detectable after 4–8 months post infection. Anti-TG antibodies occurred predominantly in aged patients in a context of a post-SARS-CoV-2-specific immune composition (R2 = 0.31; p = 0.044). This study shows that increased anti-CCP and anti-TG autoantibody levels can remain long-term after recovering even from mildly experienced COVID-19. The inter-relationship of the lung as viral entry side and RA- and CD-associated autoimmunity indicates that a SARS-CoV-2-infection could be a relevant environmental factor in their pathogenesis.

Objective: Coagulation and fibrinolysis are interrelated with the expression of vascular endothelial growth factor (VEGF), which frequently is increased in axial spondyloarthritis (axSpA). We tested whether (i) α2-antiplasmin (A2AP) Arg6Trp, (ii) fibrinogen, factor XIII A-subunit or B-subunit genotypes are associated with VEGF levels and assessed whether the known association between elevated VEGF and radiographic spinal progression in axSpA depends on genetic background. Methods: One hundred and eighty-six axSpA patients from the German Spondyloarthritis Inception Cohort were genotyped, characterized for VEGF levels, and statistically analyzed. The association between VEGF and radiographic spinal progression was assessed in dependence on genetic background in stratified analyses. Results: A2AP 6Trp carriage was associated with VEGF elevation (OR: 2.37, 95% CI: 1.06–5.29) and VEGF levels (6Trp, 455 ± 334 pg/mL; 6Arg/Arg, 373 ± 293 pg/mL; p < 0.008). Association between elevated VEGF and radiographic spinal progression in axSpA (OR: 3.11, 95% CI: 1.02–8.82) depended remarkably on the fibrinogen (FGA) genotype. When considering axSpA patients with elevated VEGF, in FGA rs6050A>G wild types, 42.1% of patients (8 of 19) progressed, while in G-allele carriers, no radiographic progression happened (0 of 13) (p < 0.04). Conclusions: The A2AP Arg6Trp genotype seems to influence VEGF levels in axSpA. The predictive value of VEGF elevations in respect of radiographic spinal progression in axSpA depends on FGA genotypes.

Additional file 3. Changes on standard blood counts and the cellular immune status after continuous and interval exercise. Peripheral blood samples of healthy donors (n = 12) were analysed by flow cytometry at different time points before and after a single 30 min continuous (CONT) or interval (HIT) exercise (before, directly after, 1 h after and 24 h after exercise). Determined frequencies of CD45+ leukocytes, CD14+ monocytes, CD19+ B cells, CD3+/CD56+ NKT cells, and CD3−/CD56+ NK cells with their bright and dim subsets are displayed as normalized frequencies. Data are mean ± SD.

Additional file 4. Changes in standard blood counts after continuous and interval exercise differentiating between moderately and highly fit donors. Peripheral blood samples of healthy donors (n = 12) were analysed using a haemocytometer at different time points before and after a single 30 min continuous (CONT) or interval (HIT) exercise (before, directly after, 1 h after and 24 h after exercise). Donors were grouped according to their fitness level (moderate, n = 8, and high, n = 4, tested with the International Physical Activity Questionnaire, IPAQ). (A) Lymphocyte, granulocyte, and monocyte frequency is shown as percentage of the total leukocyte count. (B) White blood cell (WBC), lymphocyte, granulocyte, and monocyte counts are shown as absolute counts (×103/µl blood). Data are shown as mean ± SD.

Additional file 7. Effects of a single continuous and interval exercise on CD8+ T-cell cholesterol and activation levels in a donor-related setting. Peripheral blood samples of healthy donors (n = 5) were analysed at different time points before and after a single 30 min continuous (CONT) or interval (HIT) exercise (before, directly after, 1 h after and 24 h after exercise) to (A) quantify the level of cholesterol (µM)/1 × 106 cells (Amplex Red Cholesterol Assay Kit) and (B) to obtain the expression of CD69 on CD8+ T cells (flow cytometry). Data are shown in a donor-related setting as mean ± SD. Statistically significant difference is indicated by (*p

Additional file 1. Representative flow cytometric plots. Representative flow cytometry plots illustrating the gating strategy used to analyse (A) the cellular immune status in whole blood sample on a single-cell platform using TruCount tubes, (B) the enrichment efficiency of IFN-γ+ VSTs selected from PBMCs using the IFN-γ cytokine secretion assay, and (C) the activation level of isolated CD8+ T cells using CD69 as specific marker before and after exercise.

Additional file 5. Effects of a single continuous and interval exercise on stress, proliferation markers, T-cell cytotoxicity and parameters of cellular age. Peripheral blood samples of healthy donors (n = 12) were analysed at different time points before and after a single 30 min continuous (CONT) or interval (HIT) exercise (before, directly after, 1 h after and 24 h after exercise) using real-time PCR for quantification of (A) HSP70- and Ki67-, IFN-γ-, and granzyme B-mRNA levels. Constitutively expressed GAPDH gene was used as the reference standard for normalization of mRNA levels. RQ values were calculated by the delta–delta CT method. (B) Blood samples were further analysed regarding p16 levels, telomere ratio and telomere base pairs. Results are displayed as RQ, with the value from before exercise being the base line, and shown as mean ± SD.

Additional file 6. Impact of a single continuous and interval exercise on antigen-specific T-cell responses. Peripheral blood samples of healthy donors (n = 12) were analysed separately with the two-sample t approach adjusting for period at different time points before and after a single 30 min continuous (CONT) or interval (HIT) exercise (before, directly after, 1 h after and 24 h after exercise). Isolated PBMCs were stimulated overnight with CMV-, EBV- and AdV-specific peptide pools (CMV pp65, CMV IE1, EBV EBNA1, and EBV Consensus, AdV5 Hexon, and AdV5 Penton) and frequencies of functional-active virus-specific T cells were determined by IFN-γ EliSpot assay as spots per 1000 CD3+ T cells

Introduction: Serologic studies are crucial for clarifying the regional dynamics of the SARS-CoV-2 coronavirus pandemic as well as the success of a vaccination campaign against COVID-19. We describe a cohort study investigating the seroprevalence of antibodies against SARS-CoV-2 in Magdeburg (Saxony-Anhalt, Germany). Protocol and study design: The SeMaCo study (Serologische Untersuchungen bei Blutspendern des Großraums Magdeburg auf Antikörper gegen SARS-CoV-2) is a longitudinal, regional cohort study to assess the seroprevalence of COVID-19 in blood donors from Magdeburg (Capital of Saxony-Anhalt) and surrounding areas. We consider blood donors as a surrogate for the healthy, working-age population of Saxony-Anhalt. The study primarily aims to measure the prevalence and kinetics of IgG antibodies against SARS-CoV-2 in first time and repeat blood donors over a period of 21 months. The study explores four survey periods of three to four months each (January–April 21, July–October 21, February–April 22, July–October 22). At each visit, we will assess the attitude towards vaccination, the antibody response following vaccination, as well as undesired vaccination effects. Furthermore, we will collect data on occupational activities, housing conditions and the frequency of family and other social contacts. Discussion: The SeMaCo study extends the spectrum of seroepidemiological investigations in Germany. A longitudinal observation with repeated testing and serial interviews can provide a more accurate view on the dynamics of COVID-19 prevalence and spread than repeated cross-sectional studies. Based on interim results from similar studies, we expect a seroprevalence of SARS-CoV-2 antibodies below 5% in the first survey period. SeMaCo will influence policy decisions and preventative measures.

ZusammenfassungDaratumumab ist ein humaner, monoklonaler Anti-CD38-Antikörper, der zur Therapie des multiplen Myeloms eingesetzt wird. CD38 ist ein Oberflächenmolekül, das auf einer Reihe von Zellen und Geweben exprimiert wird, auch auf Erythrozyten. Nach Verabreichung von Daratumumab kommt es regelmäßig zur Interferenz im indirekten Antihumanglobulintest (IAT). In der Folge werden alle auf dem IAT basierenden immunhämatologischen Testverfahren (serologische Verträglichkeitsproben, Antikörpersuchtests, Antikörperdifferenzierungen) unspezifisch positiv. Die Daratumumab-Interferenz kann bis zu 6 Monate nach Ende der Behandlung auftreten. Das aktuelle Standardverfahren mit Dithiothreitol (DTT) zur Behebung der Interferenz ist mit Nachteilen verbunden (unter anderem hoher Zeitbedarf, Beeinträchtigung der Erythrozytenmembran mit Zerstörung bestimmter Blutgruppen und Hämolyse). Wir stellen hier einen neuen, einfachen und zeitsparenden Ansatz zur Vermeidung der Daratumumab-Interferenz vor, der auch den mit DTT verbundenen Erythrozytenschaden vermeidet.

Aims Intracoronary infusion of autologous nucleated bone marrow cells (BMCs) enhanced the recovery of left ventricular ejection fraction (LVEF) after ST-segment elevation myocardial infarction (STEMI) in the randomised-controlled, open-label BOOST trial. We reassessed the therapeutic potential of nucleated BMCs in the randomised placebo-controlled, double-blind BOOST-2 trial conducted in 10 centres in Germany and Norway. Methods and results Using a multiple arm design, we investigated the dose-response relationship and explored whether γ-irradiation which eliminates the clonogenic potential of stem and progenitor cells has an impact on BMC efficacy. Between 9 March 2006 and 16 July 2013, 153 patients with large STEMI were randomly assigned to receive a single intracoronary infusion of placebo (control group), high-dose (hi)BMCs, low-dose (lo)BMCs, irradiated hiBMCs, or irradiated loBMCs 8.1 ± 2.6 days after percutaneous coronary intervention (PCI) in addition to guideline-recommended medical treatment. Change in LVEF from baseline (before cell infusion) to 6 months as determined by MRI was the primary endpoint. The trial is registered at Current Controlled Trials (ISRCTN17457407). Baseline LVEF was 45.0 ± 8.5% in the overall population. At 6 months, LVEF had increased by 3.3 percentage points in the control group and 4.3 percentage points in the hiBMC group. The estimated treatment effect was 1.0 percentage points (95% confidence interval, -2.6 to 4.7; P = 0.57). The treatment effect of loBMCs was 0.5 percentage points (-3.0 to 4.1; P = 0.76). Likewise, irradiated BMCs did not have significant treatment effects. BMC transfer was safe and not associated with adverse clinical events. Conclusion The BOOST-2 trial does not support the use of nucleated BMCs in patients with STEMI and moderately reduced LVEF treated according to current standards of early PCI and drug therapy.

Adenovirus (HAdV) infections confer a high risk of morbidity and mortality for immunocompromised patients after stem cell transplantation (SCT). Treatment with standard antiviral drugs is of limited efficacy and associated with a high rate of adverse effects. HAdV-specific T cells are crucial for sustained viral elimination and the efficacy of adoptive T-cell therapy with donor-derived HAdV-specific T cells has been reported by several investigators. Here, we report our experience with the transfer of HAdV-specific T cells specific for penton, which was recently identified as an immunodominant target of T cells, and hexon in a 14-year-old boy after T-cell-depleted haploidentical SCT for myelodysplastic syndrome (MDS). He developed severe HAdV-associated enteritis complicated by acute graft-versus-host disease (GvHD). The patient received ten infusions of allogeneic HAdV-specific T cells manufactured from the haploidentical stem cell donor using the CliniMacs Interferon-γ (IFN-γ) cytokine capture and immunomagnetic selection. Initially, T cells were generated against the immunodominant target hexon and in subsequent transfers dual antigen-specific T cells against hexon and penton were applied. T-cell transfers were scheduled individually tailored to current immunosuppressive treatment. Each transfer was followed by reduction of HAdV load in peripheral blood and clinical improvement. Importantly, T-cell responses to both penton and hexon pools emerged in patient blood after repetitive transfers. Unfortunately, the patient experienced bacterial sepsis, and in this context, severe GvHD requiring intensive immunosuppression followed by secondary progression of HAdV infection. The patient succumbed to multiorgan failure 283 days after SCT. This case demonstrates the feasibility of HAdV-specific T-cell transfer even in the presence of immunosuppressive treatment. Targeting of multiple immunodominant viral proteins may prove valuable in patients with complicated HAdV infections.

Background Adoptive transfer of virus-specific T cells (VSTs) represents a prophylactic and curative approach for opportunistic viral infections and reactivations after transplantation. However, inadequate frequencies of circulating memory VSTs in the T-cell donor’s peripheral blood often result in insufficient enrichment efficiency and purity of the final T-cell product, limiting the effectiveness of this approach. Methods This pilot study was designed as a cross-over trial and compared the effect of a single bout (30 min) of high-intensity interval training (HIT) with that of 30 min of continuous exercise (CONT) on the frequency and function of circulating donor VSTs. To this end, we used established immunoassays to examine the donors’ cellular immune status, in particular, with respect to the frequency and specific characteristics of VSTs restricted against Cytomegalovirus (CMV)-, Epstein–Barr-Virus (EBV)- and Adenovirus (AdV)-derived antigens. T-cell function, phenotype, activation and proliferation were examined at different time points before and after exercise to identify the most suitable time for T-cell donation. The clinical applicability was determined by small-scale T-cell enrichment using interferon- (IFN-) γ cytokine secretion assay and virus-derived overlapping peptide pools. Results HIT proved to be the most effective exercise program with up to fivefold higher VST response. In general, donors with a moderate fitness level had higher starting and post-exercise frequencies of VSTs than highly fit donors, who showed significantly lower post-exercise increases in VST frequencies. Both exercise programs boosted the number of VSTs against less immunodominant antigens, specifically CMV (IE-1), EBV (EBNA-1) and AdV (Hexon, Penton), compared to VSTs against immunodominant antigens with higher memory T-cell frequencies. Conclusion This study demonstrates that exercise before T-cell donation has a beneficial effect on the donor’s cellular immunity with respect to the proportion of circulating functionally active VSTs. We conclude that a single bout of HIT exercise 24 h before T-cell donation can significantly improve manufacturing of clinically applicable VSTs. This simple and economical adjuvant treatment proved to be especially efficient in enhancing virus-specific memory T cells with low precursor frequencies.

Background: Currently, there is an extensive but highly inconsistent body of literature regarding donor adverse events (AEs) in haemapheresis. As the reports diverge with respect to types and grading of AEs, apheresis procedures and machines, the range of haemapheresis-related AEs varies widely from about 0.03% to 6.6%. Methods: The German Society for Transfusion Medicine and Immunohaematology (DGTI) formed a ‘Haemapheresis Vigilance Working Party' (Arbeitsgemeinschaft Hämapheresevigilanz; AGHV) to create an on-line registry for comprehensive and comparable AE assessment with all available apheresis devices in all types of preparative haemapheresis: plasmapheresis (PLS), plateletpheresis (PLT), red blood cell apheresis, all kind of leukaphereses (autologous/allogeneic blood stem cell apheresis, granulocyte apheresis, lymphocyte/monocyte apheresis) and all possible types of multi-component apheresis. To ensure the comparability of the data, the AGHV adopted the ‘Standard for Surveillance of Complications Related to Blood Donation' from the International Society for Blood Transfusion in cooperation with the International Haemovigilance Network (IHN) and the American Association of Blood Banks for AE acquisition and automated evaluation. The registry is embedded in a prospective observational multi-centre study with a study period of 7 years. Results: A preliminary evaluation encompassed the time period from January, 2012 to December, 2015. During this time, the system proved to be safe and stable. Out of approximately 345,000 haemaphereses 16,477 AEs were reported (4.9%) from 20 participating centres. The majority of AEs occurred in PLSs (63%), followed by PLT (34.5%) and SC (2.2%). Blood access injuries (BAI) accounted for about 55% of the supplied AEs, whereas citrate toxicity symptoms, vasovagal reactions and technical events (e.g. disposable leakages, software failures) rather equally affected haemaphereses at 8-15%. Out of 12,348 finalized AEs, 8,759 (70.1%) were associated with a procedure-related break-off, with BAI being the prevailing cause (5,463/8,759; 62.4%). An automated centre- and procedure-specific AE evaluation according to the latest IHN standard and AGHV pre-settings is available within a few minutes. Conclusions: An on-line electronic platform for comprehensive assessment and centre-specific automated evaluation of AEs in haemaphereses has been developed and proved to be stable and safe over a period of 4 years.

Abnormal transcranial Doppler velocities (TCD) indicate an increased risk of stroke in patients with sickle cell anemia (SCA) and require regular blood transfusions. Hematopoietic stem cell transplantation (HSCT) is under discussion as an alternative to chronic transfusion in these patients. This retrospective analysis includes 9 patients with SCA undergoing HSCT at a single center in Germany. Special focus was given to the neurologic follow-up and to the results of TCD studies. High risk of stroke or previous stroke was an HSCT-indication in 8 of 9 patients, although most patients had more than one indication for HSCT. TCD was normalized in all 5 patients after HSCT in whom this test was available. None of the patients developed a stroke after HSCT. No further strokes occurred even in patients that experienced recurrent strokes during chronic transfusion before HSCT. 2 of the 9 patients received a 10/10 HLA-matched unrelated donor graft, the others matched related grafts. All patients were alive, free of SCA symptoms and transfusion-independent with stable chimerism 3–11 years after HSCT. Pulmonary function tests normalized in 1 patient with severe sickle cell lung disease. HSCT is able to prevent stroke in patients with SCA. Its perspectives and limitations should be discussed early during the treatment of a patient with complicated SCA.

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 and aims. The infusion of enriched CMV-specific donor T-cells appears to be a suitable alternative for the treatment of drug resistant CMV reactivation or de novo infection after both solid organ and hematopoietic stem cell transplantation. Antiviral lymphocytes can be selected from apheresis products using the CliniMACS Cytokine-Capture-System® either with the well-established CliniMACS® Plus (Plus) device or with its more versatile successor CliniMACS Prodigy® (Prodigy). Methods. Manufacturing of CMV-specific T-cells was carried out with the Prodigy and Plus in parallel starting with 0.8-1*109 leukocytes collected by lymphapheresis (n=3) and using the MACS GMP PepTivator® HCMVpp65 for antigenic re-stimulation. Target and non-target cells were quantified by a newly developed single-platform assessment and gating strategy using positive (CD3/CD4/CD8/CD45/IFN-gamma), negative (CD14/CD19/CD56), and dead cell (7-AAD) discriminators. Results. Both devices produced largely similar results for target cell viabilities: 37.2%-52.2% (Prodigy) vs. 51.1%-62.1% (Plus) CD45+/7-AAD- cells. Absolute numbers of isolated target cells were 0.1-3.8*106 viable IFN-gamma+ CD3+ cells. The corresponding proportions of IFN-gamma+ CD3+ cells ranged between 19.2% and 95.1% among total CD3+ cells and represented recoveries of 41.9%-87.6%. Within two parallel processes predominantly IFN-gamma+ CD3+CD8+ cytotoxic T-cells were enriched compared to one process that yielded a higher amount of IFN-gamma+ CD3+CD4+ helper T lymphocytes. T-cell purity was higher for the Prodigies products that displayed a lower content of contaminating IFN-gamma- T-cells (3.6%-20.8%) compared to the Plus products (19.9%-80.0%). Conclusions. The manufacturing process on the Prodigy saved both process and hands-on time due to its higher process integration and ability for unattended operation. Although the usage of both instruments yielded comparable results, the lower content of residual IFN-gamma- T-cells in the target fractions produced with the Prodigy may allow for a higher dosage of CMV-specific donor T-cells without increasing the risk for graft versus host disease.

BACKGROUND: The objective was to investigate potential risks for apheresis donors associated with a triple-plateletpheresis (TP) program. STUDY DESIGN AND METHODS: Eleven hemapheresis centers randomly assigned 411 repeat donors (ratio, 1:1.2) to either double plateletpheresis (DP; 185 donors) or TP (226 donors) with a platelet (PLT) target content of at least 5.0 × 1011 PLTs/DP and at least 7.5 × 1011 PLTs/TP. The primary endpoint was procedure-related postapheresis PLT count of at least 150 × 109/L (probability, ≥98%). Secondary endpoints were apheresis characteristics and donor adverse reactions. RESULTS: In 6 of 1133 DPs (0.5%) in 4 of 185 donors (2.2%) and in 20 of 1020 TPs (2.0%) in 14 of 226 donors (6.2%), postapheresis PLT counts were below 150 × 109/L. There were marginal but significant differences in collection efficiency (DP, 69.2 ± 9.1%; TP, 70.9 ± 9.0%; p ≤ 0.0001) and collection rate (DP, 10.4 × 109 ± 2.3 × 109 PLTs/min; TP, 10.8 × 109 ± 2.3 × 109 PLTs/min; p ≤ 0.005). The PLT yields were 5.9 × 1011 ± 0.8 × 1011 PLTs for DP and 8.3 × 1011 ± 0.9 × 1011 PLT for TP (p ≤ 0.0001) at processing times of 59 ± 13 minutes (DP) versus 80 ± 16 minutes (TP; p ≤ 0.0001). Significant PLT recruitment (1.10 ± 0.14 vs. 1.20 ± 0.23; p

The reactivation of the human cytomegalovirus (CMV) can be prevented or controlled by the adoptive transfer of ex vivo expanded donor-derived CMV-specific T lymphocytes. Several methods for expansion and adoptive transfer of CMV-specific T cells have been developed using either defined CMV peptides or peptide pools for antigen-specific T-cell stimulation. The majority of studies have focused on the lower matrix protein (pp65) and the immediate-early protein-1 (IE-1) of CMV as immunodominant targets. We investigated the behavior of secretory CMVpp65 (sCMVpp65) with respect to its capacity to stimulate pp65-specific T cells independently of human lymphocyte antigen (HLA) type and even in donors unresponsive to the immunodominant HLA-A*0201-restricted CMVpp65495-503 peptide. To facilitate the eukaryotic expression and isolation procedures, we constructed an HLA-A*0201/CMVpp65 fusion protein that is secreted into the supernatant of human embryonic kidney 293 (HEK293) cells. CMV-specific CD4 and CD8 T cells generated by culturing unfractionated peripheral blood mononuclear cells in the presence of recombinant sCMVpp65 did not differ in function with regard to cytotoxicity and interferon-gamma (IFN-gamma) production compared with cytotoxic T cells induced using the well-studied HLA-A*0201-restricted CMVpp65495-503 peptide. We demonstrated that polyclonal CMV-specific T cells could be generated from CMV-seropositive individuals expressing HLA alleles for which no immunogenic epitopes have been identified so far. The production of recombinant sCMVpp65 can easily be adapted to good manufacturing practice conditions and can be used to generate large numbers of immunogenic pathogen-derived proteins for therapeutic applications.

BACKGROUND: Delayed donor red blood cell chimerism (DRCC), pure red blood cell aplasia (PRCA), and autoimmune hemolytic anemia (AIHA) are poorly documented complications after hematopoietic cell transplantation (HCT). The clinical variable “prolonged isolated red blood cell transfusion requirement” (PRTR) was evaluated as a trigger for an extended diagnostic workup. STUDY DESIGN AND METHODS: PRTR was defined as the need for red blood cell (RBC) transfusions beyond Day 60 after HCT. We analyzed 487 patients transplanted between 2000 and 2006. Median age was 37 years (range, 0-70 years). Peripheral blood stem cells (n = 344), marrow (n = 138), and cord blood (n = 5) from 278 unrelated and 209 family donors were used. RESULTS: Univariate analysis identified age (incidence of 18.3% among elderly patients, 10.5% in adults, and 2.0% among children [p = 0.002]), ABO incompatibility (16.4% after major incompatible, 2.9% after minor incompatible, and 9.4% after ABO-compatible transplantations [p = 0.003]), conditioning (15.2% after reduced-intensity regimens vs. 7.3% after myeloablative conditioning; p = 0.006), donor type (13.2% after HLA-matched unrelated, 13.6% after mismatched unrelated, 5.7% after matched related, and 0.0% after mismatched related grafts; p = 0.026), and acute graft-versus-host disease (aGVHD; 7.1% with aGVHD vs. 12.5% without aGVHD; p = 0.046) as predisposing factors. In multivariate analysis minor ABO incompatibility (odds ratio [OR] = 0.2, p = 0.01), younger age (OR = 0.1, p = 0.02), and matched related HCT (OR = 0.4, p = 0.02) remained independent protective factors. CONCLUSIONS: PRTR could serve as a trigger for a standardized screening for DRCC, PRCA, and AIHA after HCT.

We compare the actual with the potential donor exposure and possible infection rates in the Hanover Medical School (MHH) platelet (PLT) transfusion recipients if the current MHH standard of apheresis PLT concentrate (A-PC) supply would be replaced by a pooled PLT concentrate (P-PC) transfusion regimen.The electronic records of the MHH Institute of Transfusion Medicine and the MHH Department of Medical Controlling were evaluated to assess the development of PLT needs and supply at MHH from 2003-2006. For 2006, we evaluated all PLT transfusion recipients with respect to their overall transfusion needs, classified them for low and high PLT transfusion needs, and related them to the diagnostic groups that underlie their PLT demands. We assumed a P-PC preparation procedure using 4 whole blood-derived buffy coats for all calculations for potential donor exposure. To predict the possible infection rates of an unrecognized viral infection with low prevalence in the general population to A-PC or to P-PC recipients and the influence of neutralizing agent specific antibodies (NAB), we established a mathematical contamination/infection model based on the current PLT transfusion mode and data about GBV-C virus infection among Hanover blood donors.From 2003 to 2006, the 1,300-1,400 persons comprising MHH apheresis donor pool covered a 36% increase in PC transfusions. The exclusive use of P-PCs instead of A-PC would require a total of 36,240-49,276 whole blood donations to meet MHH demands, corresponding to a more than 1 log step increase in donor exposure. For individual hematological patients, the change to P-PCs would imply an 80-125%, for individual surgical patients a 40-50% higher donor exposure. Our infection model revealed an approximately 4 times higher infection.A change to P-PC would imply a more than one log step higher donor exposure, and an unrecognized infection with a prevalence around 1% leads to an up to 4 times higher infection rate. A general change in the PC transfusion policy that favors P-PCs is dangerous and must be avoided.Ziel der Studie war die Erfassung der tatsächlichen im Vergleich zur potentiellen Spenderexposition sowie der möglichen Infektionsraten bei Empfängern von Thrombozytenkonzentraten (TK), falls der gegenwärtige Transfusionsstandard an der Medizinischen Hochschule Hannover (MHH) – die Patientenversorgung mit Apheresekonzentraten (A-TK) – von einer Transfusionsstrategie mit gepoolten Thrombozytenkonzentraten (P-TK) abgelöst würde.Wir haben die elektronischen Datenbanken des MHH-Instituts für Transfusionsmedizin sowie der MHH-Abteilung für Medizinkontrolling bezüglich der Entwicklung des Thrombozytenbedarfs und der Thrombozytenversorgung für die Jahre 2003–2006 ausgewertet. Bezogen auf das Jahr 2006 wurden alle MHH-Thrombozytenempfänger hinsichtlich des Gesamttransfusionsbedarfs, niedrigen bzw. hohen Thrombozytenbedarfs bzw. auf die dem Thrombozytenbedarf zugrunde liegenden Erkrankungen untersucht. Auf der Basis von 4 Buffy Coats aus Vollblutkonserven als Ausgangsmaterial für die Herstellung eines P-TK berechneten wir die potentielle Spenderexposition für die MHH-TK-Empfänger. Wir entwickelten ein mathematisches Modell, um mögliche Infektionsraten bei einer unerkannten viralen Infektion mit niedriger Prävalenz in der Allgemeinbevölkerung bei A-TK-oder bei P-TK-Empfängern unter Einschluss neutralisierender virusspezifischer Antikörper vorherzusagen. Dieses Modell basiert auf der gegenwärtigen Transfusionspraxis für TK in Hannover und bekannten Daten zu GBV-C-Virusinfektionen bei Hannoveraner Blutspendern.Der 1,300–1,400 Personen umfassende MHH-Apheresespenderpool stellte für die Jahre 2003 bis 2006 sicher, dass der 36%ige Anstieg der Thrombozytentransfusionen bewältigt werden konnte. Die ausschlieβliche Anwendung von P-TK anstelle von A-TK hätte 36.240–49.276 Vollblutspender erfordert, was einem Anstieg der Spenderexposition um mehr als eine Logstufe entsprochen hätte. Individuelle Thrombozytenempfänger mit niedrigem, moderatem, hohem und sehr hohem A-TK-Bedarf hätten eine um 42, 67, 84 und 109% höhere Spenderexposition gehabt. Für individuelle hämatologische Patienten hätte der Wechsel des Transfusionsregimes zu P-TK eine um 80–125%, für individuelle chirurgische Patienten eine um 40–50% höhere Spenderexposition zur Folge gehabt. Unser Rechenmodell zur Abschätzung von Infektionsfolgen zeigt eine zirka viermal höhere Infektionsrate.Ein Wechsel der Transfusionsstrategie auf Pool-TK würde mit einer um mehr als eine Log-Stufe erhöhten Spenderexpositition einhergehen, und eine unerkannte Infektion mit einer Prävalenz von zirka 1% führt zu einer um den Faktor 4 erhöhten Infektionsrate bei Pool-TK-Empfängern. Ein genereller Wechsel in der Transfusionsstrategie für Thrombozytenkonzentrate ist daher als gefährlich anzusehen und muss unbedingt vermieden werden.

Intracoronary application of BM-derived cells for the treatment of acute myocardial infarction (AMI) is currently being studied intensively. Simultaneously, strict legal requirements surround the production of cells for clinical studies. Thus good manufacturing practice (GMP)-compliant collection and preparation of BM for patients with AMI was established by the Cytonet group.As well as fulfillment of standard GMP requirements, including a manufacturing license, validation of the preparation process and the final product was performed. Whole blood (n=6) and BM (n=3) validation samples were processed under GMP conditions by gelafundin or hydroxyethylstarch sedimentation in order to reduce erythrocytes/platelets and volume and to achieve specifications defined in advance. Special attention was paid to the free potassium (6 mmol/L), some rheologically relevant cellular characteristics (hematocrit0.45, platelets450 x 10(6)/mL) and the sterility of the final product.The data were reviewed and GMP compliance was confirmed by the German authorities (Paul-Ehrlich Institute). Forty-five BM cell preparations for clinical use were carried out following the validated methodology and standards. Additionally three selections of CD34+ BM cells for infusion were performed. All specification limits were met. Discussion In conclusion, preparation of BM cells for intracoronary application is feasible under GMP conditions. As the results of sterility testing may not be available at the time of intracoronary application, the highest possible standards to avoid bacterial and other contaminations have to be applied. The increased expense of the GMP-compliant process can be justified by higher safety for patients and better control of the final product.

Clinically the diagnosis of an autoimmune hemolytic anemia (AIHA) often starts with a patient with an unclear anemia, while in the laboratory the first hint is a blood sample with a positive direct antiglobulin test (DAT). Both anemia and positive DAT are non-specific findings that raise many questions. These include a wide range of laboratory characteristics such as immunoglobulin (sub-)class of the red blood cell (RBC) autoantibodies (Au-ab), presence of warm or cold Au-ab, thermal range of cold Au-ab, capability of Au-ab to activate complement, specificity of warm or cold Au-ab, and masking of additional RBC alloantibodies by Au-ab. Moreover, the clinical findings have to be scrutinized critically: Was there a sudden onset or a gradual beginning of hemolytic anemia? Is the hemolysis life-threatening? Does the patient suffer from a persisting systemic autoimmune or tumor disease, in particular a hematological malignancy? Has the patient been transfused within a certain time span before the first occurrence of the anemia symptoms? Was there a recent infection? Does the patient take drugs, newly or regularly? If a clinician is confronted with a newly diagnosed AIHA, each of these questions has to be answered to distinguish ‘idiopathic AIHA’ as a disorder of its own from ‘secondary AIHA’ as a symptom of a detectable underlying disease (e.g. chronic lymphocytic leukemia, systemic lupus erythematosus, Mycoplasma pneumoniae infection). This classification is important for choosing the appropriate medical treatment of AIHA. In idiopathic AIHA that is present in about 45% of the AIHA cases [1] immunosuppressive drugs (corticosteroids and others) are most frequently applied to cope with severe hemolysis [2]. In secondary AIHA the ‘drug of choice’ is governed by the underlying disease and may range from ‘wait and see’ in self-limiting cases of infection-related AIHA, over immunosuppressive drugs in cases with an overt systemic autoimmune disorder to chemotherapy in patients with lymphoproliferative malignancies [2,3]. Regardless of the clinical background, many AIHA patients have to be transfused, at least temporarily, as none of the available drugs are able to stop severe hemolysis immediately [4]. In view of these facts, each newly diagnosed ‘AIHA’ may become a challenging experience for the doctor who is responsible for the patient for several reasons: i) There is no close relationship between the laboratory findings and the clinical symptoms of the patient. ii) Hypoxic anemia may require urgent transfusion. iii) A deliberate search for an underlying disease often leads to the somewhat dissatisfactory ‘idiopathic’ AIHA. Moreover, none of the administered drugs - neither corticosteroids that are in routine use for decades nor newly introduced monoclonal antibodies such as rituximab - are always effective, and their beneficial immediate or long-term effects are not safely predictable [5,6]. The present issue of Transfusion Medicine And Hemotherapy is an attempt to summarize and review theories, ideas, and medical expert knowledge on AIHA. John Freedman [7] and Wilma Barcellini [8] present the history of AIHA and discuss the current concepts that contribute to RBC destruction. Barcellini reports on the roles of cytokines and cytotoxicity effector cells (CD8+ T cells, natural killer cells, and activated macrophages) in the AIHA disease process, thereby explaining the great clinical heterogeneity of AIHA ranging from a fully compensated (‘uneventless') clinical course to rapidly evolving severe life-threatening hemolysis. The contributions of Sigbjorn Berentsen [9] and Thilo Bartolmas et al. [10] deal with the role of the complement system in the development of AIHA and describe the main mediators of complement-driven hemolysis, in particular in cold Au-ab-associated AIHA. Charles H. Packman [11] gives an overview on the clinical manifestations of AIHA while Beate Mayer et al. [12] present more than 70 cases with drug-induced hemolysis, mainly as a result of diclofenac, piperacillin and cephalosporin ingestion. Abdulgabar Salama [13] reviews pitfalls that may hamper the diagnose ‘AIHA’ and summarizes his professional experience with the medical treatment of primary (‘idiopathic') AIHA over a period of 30 years. The latter contribution might provoke intense discussions among transfusion medicine specialists as the author attenuates the therapeutic enthusiasm with regard to newer drugs such as rituximab as the new ‘gold standard’ for medical AIHA treatment. When looking at the contents of this issue, you might wonder why one name seems to be nearly omnipresent in this special topic. It is not the task of this editorial to fully appreciate the tremendous scientific work of Abdulgabar Salama in the field of immunohematology over the last 30 years. However, as obvious for every specialist in transfusion medicine, Abdulgabar Salama has not only significantly influenced the field but has clearly dominated (together with his Californian colleague George Garratty) the diagnosis and the pathogenic background of drug-induced AIHA. His most important contribution is the discovery that many drugs supposed to be involved in severe AIHA are detectable in laboratory investigations only when their degradation products, either in serum/plasma or urine, are available (e.g. nomifensine [7], cephalosporin antibiotics such as ceftriaxone or cefotaxime [8,9], or the non-steroid antirheumatic diclofenac [10], just to mention some frequently used medications). Another similarly important contribution to the understanding of the pathogenicity of AIHA is his observation that transfusion itself is able to induce Au-ab formation, often in coexistence with transfusion-associated alloantibody formation, thereby complicating a delayed serologic/hemolytic transfusion reaction [11]. These are only a few reasons why we are very glad to have allured such an exposed representative of AIHA research to significantly contribute to this special topic.