Yara Strehler, N. Lachmann, M. Niemann, F. Halleck, K. Budde, Axel Pruß
Introduction: Eurotransplant established the acceptable mismatch (AM) program to facilitate timely kidney transplantations of highly sensitized patients, but long-term granular clinical and immunological outcomes regarding overall graft survival and de novo DSA (dnDSA) formation are still intensively researched. The right choice of induction therapy in patients with differing immunological risk is not conclusively determined, as well as the impact of human leukocyte antigen (HLA) epitope matching on dnDSA formation. Methods: This monocentric, retrospective study analyzed 94 patients transplanted within the AM program between 2000 and 2019 compared to case-control matched cohorts of non- (PRA 0–5%; PRA-0) and intermediately sensitized (PRA 6–84%; PRA-6/84) patients transplanted through Eurotransplant Kidney Allocation System. Results: Estimated 10-year overall graft survival between the PRA-0 and AM cohorts was similar, whereas PRA-6/84 was significantly disadvantageous compared to PRA-0. Estimated 10-year incidence of antibody-mediated rejection rates was significantly lower in the PRA-0 group compared to AM and PRA-6/84 groups. Compared to the AM group, estimated incidence of de novo donor-specific antibody (dnDSA) was significantly lower in PRA-0 patients, with no differences between the AM and PRA-6/84 cohorts. The PRA-6/84 cohort was the only subgroup in which interleukin-2 receptor antagonist (IL2RA) induction was associated with longer overall graft survival, patient survival, and graft survival compared to depleting induction (ATG or OKT3). Broad HLA-A, -B, -DR mismatches (mmABDR) and HLA epitope mismatches determined by Eplets and PIRCHE-II were predictive for dnDSA formation in the total cohort, and the AM subgroup. Discussion: The high efforts expended on AM patients are justified to allow timely organ transplantation with acceptable risk profile and non-inferior outcomes. IL2RA induction in intermediately sensitized patients is associated with superior overall graft survival, patient survival, and graft survival compared to ATG/OKT3 induction, without negative effects on rejection episodes or dnDSA formation. In silico epitope matching might further help reduce dnDSA formation, particularly in high-risk AM patients.
导言：欧洲肾移植协会（Eurotransplant）制定了可接受的错配（AM）计划，以促进高度致敏患者及时进行肾移植，但有关总体移植存活率和新生DSA（dnDSA）形成的长期临床和免疫学结果仍在深入研究之中。对于免疫学风险不同的患者，诱导治疗的正确选择以及人类白细胞抗原（HLA）表位匹配对 dnDSA 形成的影响尚无定论。方法：这项单中心回顾性研究分析了 2000 年至 2019 年间在 AM 计划内移植的 94 例患者，并与通过欧洲肾移植分配系统移植的非致敏（PRA 0-5%；PRA-0）和中间致敏（PRA 6-84%；PRA-6/84）患者的病例对照匹配队列进行了比较。结果：PRA-0组和AM组的估计10年总体移植物存活率相似，而PRA-6/84组与PRA-0组相比明显处于劣势。与 AM 组和 PRA-6/84 组相比，PRA-0 组的估计 10 年抗体介导排斥发生率明显较低。与 AM 组相比，PRA-0 患者的新供体特异性抗体（dnDSA）估计发生率明显较低，而 AM 组和 PRA-6/84 组之间没有差异。与去势诱导（ATG 或 OKT3）相比，PRA-6/84 组是唯一一个白细胞介素-2 受体拮抗剂（IL2RA）诱导与较长的总移植物存活率、患者存活率和移植物存活率相关的亚组。通过Eplets和PIRCHE-II确定的广义HLA-A、-B、-DR错配（mmABDR）和HLA表位错配可预测整个队列和AM亚组中dnDSA的形成。讨论：为了能在风险可接受的情况下及时进行器官移植，并获得非劣效的治疗效果，对AM患者花费大量精力是合理的。与 ATG/OKT3 诱导相比，IL2RA 诱导中间致敏患者的总体移植物存活率、患者存活率和移植物存活率更高，且对排斥反应发作或 dnDSA 的形成没有负面影响。硅表位匹配可能会进一步帮助减少 dnDSA 的形成，尤其是在高风险 AM 患者中。
{"title":"Positive Long-Term Outcome of Kidney Allocation via Acceptable Mismatch Program in Highly Sensitized Patients","authors":"Yara Strehler, N. Lachmann, M. Niemann, F. Halleck, K. Budde, Axel Pruß","doi":"10.1159/000536533","DOIUrl":"https://doi.org/10.1159/000536533","url":null,"abstract":"Introduction: Eurotransplant established the acceptable mismatch (AM) program to facilitate timely kidney transplantations of highly sensitized patients, but long-term granular clinical and immunological outcomes regarding overall graft survival and de novo DSA (dnDSA) formation are still intensively researched. The right choice of induction therapy in patients with differing immunological risk is not conclusively determined, as well as the impact of human leukocyte antigen (HLA) epitope matching on dnDSA formation. Methods: This monocentric, retrospective study analyzed 94 patients transplanted within the AM program between 2000 and 2019 compared to case-control matched cohorts of non- (PRA 0–5%; PRA-0) and intermediately sensitized (PRA 6–84%; PRA-6/84) patients transplanted through Eurotransplant Kidney Allocation System. Results: Estimated 10-year overall graft survival between the PRA-0 and AM cohorts was similar, whereas PRA-6/84 was significantly disadvantageous compared to PRA-0. Estimated 10-year incidence of antibody-mediated rejection rates was significantly lower in the PRA-0 group compared to AM and PRA-6/84 groups. Compared to the AM group, estimated incidence of de novo donor-specific antibody (dnDSA) was significantly lower in PRA-0 patients, with no differences between the AM and PRA-6/84 cohorts. The PRA-6/84 cohort was the only subgroup in which interleukin-2 receptor antagonist (IL2RA) induction was associated with longer overall graft survival, patient survival, and graft survival compared to depleting induction (ATG or OKT3). Broad HLA-A, -B, -DR mismatches (mmABDR) and HLA epitope mismatches determined by Eplets and PIRCHE-II were predictive for dnDSA formation in the total cohort, and the AM subgroup. Discussion: The high efforts expended on AM patients are justified to allow timely organ transplantation with acceptable risk profile and non-inferior outcomes. IL2RA induction in intermediately sensitized patients is associated with superior overall graft survival, patient survival, and graft survival compared to ATG/OKT3 induction, without negative effects on rejection episodes or dnDSA formation. In silico epitope matching might further help reduce dnDSA formation, particularly in high-risk AM patients.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140417515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
U. Gravemann, W. Handke, Torsten J. Schulze, Axel Seltsam
Introduction: Bacterial contamination of blood products presumably occurs mainly during blood collection, starting from low initial concentrations of 10–100 colony-forming units (CFUs) per bag. As little is known about bacterial growth behavior and distribution in stored whole blood (WB) and WB-derived blood products, this study aims to provide data on this subject. Methods: WB units were inoculated with transfusion-relevant bacterial species (Acinetobacter baumannii, Bacillus cereus, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Pseudomonas fluorescens, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus dysgalactiae, Streptococcus pyogenes, Yersinia enterocolitica; n = 12 for each species), stored for 22–24 h at room temperature, and then centrifuged for separation into plasma, red blood cells (RBCs), and buffy coats (BCs). The latter were pooled with 3 random donor BCs and one unit of PAS-E each to yield plasma-reduced platelet concentrates (PCs). Samples for bacterial colony counting were collected after WB storage and immediately after blood component production. Sterility testing in PCs (n = 12 for each species) was performed by bacterial culture after 7 days of storage. Results: Bacterial growth in WB varied remarkably between donations and species. Streptococcus species produced the highest titers in WB, whereas Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas fluorescens did not multiply. Centrifugation resulted in preferential accumulation of bacteria in BCs, with titers of up to 3.5 × 103 CFU/mL in BCs and up to ≤0.9 × 103 CFU/mL in BC-derived PCs. Overall, 72/144 PCs (50%) tested positive for bacteria after storage. Sterility test results were species-dependent, ranging from 12 of 12 PCs tested positive for Streptococcus pyogenes to 1 of 12 PCs positive for Escherichia coli. Bacterial contamination of RBC and plasma units was much less common and was associated with higher initial bacterial counts in the parent WB units. Conclusions: Bacterial growth in WB is species-dependent and varies greatly between donations. Preferential accumulation of bacteria in BCs during manufacturing is a critical determinant of the contamination risk of BC-derived pooled PCs.
导言：血液制品中的细菌污染可能主要发生在采血过程中，最初的浓度较低，每袋只有 10-100 个菌落形成单位 (CFU)。由于人们对储存的全血（WB）和全血衍生血液制品中细菌的生长行为和分布知之甚少，本研究旨在提供这方面的数据。研究方法给 WB 单位接种与输血相关的细菌种类（鲍曼不动杆菌、蜡样芽孢杆菌、大肠埃希菌、肺炎克雷伯菌、单核细胞增生李斯特菌、荧光假单胞菌、肉毒杆菌、金黄色葡萄球菌、表皮葡萄球菌、痢疾链球菌、化脓性链球菌、小肠结肠炎耶尔森菌；n=12），在室温下保存 22-24 小时，然后离心分离成血浆、红细胞（RBC）和缓冲衣（BC）。后者与 3 个随机供体 BC 和各一个单位的 PAS-E 混合，得到血浆还原血小板浓缩物（PC）。细菌菌落计数样本在 WB 储存后和血液成分生产后立即采集。储存 7 天后，通过细菌培养对 PC（每个品种 12 个）进行无菌检测。结果不同捐赠和不同种类的 WB 中细菌生长情况差异显著。链球菌在 WB 中的滴度最高，而金黄色葡萄球菌、表皮葡萄球菌、大肠杆菌和荧光假单胞菌则不繁殖。离心后，细菌优先在 BCs 中聚集，BCs 中的滴度高达 3.5 × 103 CFU/mL，而 BC 衍生的 PCs 中的滴度则≤0.9 × 103 CFU/mL。总体而言，72/144 个 PC（50%）在储存后细菌检测呈阳性。无菌检测结果与菌种有关，化脓性链球菌阳性的 PC 有 12 例，大肠杆菌阳性的 PC 只有 1 例。红细胞和血浆单位受到细菌污染的情况要少得多，而且与母体 WB 单位中较高的初始细菌计数有关。结论白细胞计数器中细菌的生长与物种有关，不同捐赠之间差异很大。在生产过程中，细菌在 BC 中的优先积累是决定 BC 衍生的集合 PC 污染风险的关键因素。
{"title":"Growth and Distribution of Bacteria in Contaminated Whole Blood and Derived Blood Components","authors":"U. Gravemann, W. Handke, Torsten J. Schulze, Axel Seltsam","doi":"10.1159/000536242","DOIUrl":"https://doi.org/10.1159/000536242","url":null,"abstract":"Introduction: Bacterial contamination of blood products presumably occurs mainly during blood collection, starting from low initial concentrations of 10–100 colony-forming units (CFUs) per bag. As little is known about bacterial growth behavior and distribution in stored whole blood (WB) and WB-derived blood products, this study aims to provide data on this subject. Methods: WB units were inoculated with transfusion-relevant bacterial species (Acinetobacter baumannii, Bacillus cereus, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Pseudomonas fluorescens, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus dysgalactiae, Streptococcus pyogenes, Yersinia enterocolitica; n = 12 for each species), stored for 22–24 h at room temperature, and then centrifuged for separation into plasma, red blood cells (RBCs), and buffy coats (BCs). The latter were pooled with 3 random donor BCs and one unit of PAS-E each to yield plasma-reduced platelet concentrates (PCs). Samples for bacterial colony counting were collected after WB storage and immediately after blood component production. Sterility testing in PCs (n = 12 for each species) was performed by bacterial culture after 7 days of storage. Results: Bacterial growth in WB varied remarkably between donations and species. Streptococcus species produced the highest titers in WB, whereas Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas fluorescens did not multiply. Centrifugation resulted in preferential accumulation of bacteria in BCs, with titers of up to 3.5 × 103 CFU/mL in BCs and up to ≤0.9 × 103 CFU/mL in BC-derived PCs. Overall, 72/144 PCs (50%) tested positive for bacteria after storage. Sterility test results were species-dependent, ranging from 12 of 12 PCs tested positive for Streptococcus pyogenes to 1 of 12 PCs positive for Escherichia coli. Bacterial contamination of RBC and plasma units was much less common and was associated with higher initial bacterial counts in the parent WB units. Conclusions: Bacterial growth in WB is species-dependent and varies greatly between donations. Preferential accumulation of bacteria in BCs during manufacturing is a critical determinant of the contamination risk of BC-derived pooled PCs.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140444257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lina S. Silva-Bermúdez, D. Heidenreich, Stefan A. Klein, Patrick Wuchter, Harald Klüter, Sabine Kayser
Introduction: Major ABO-incompatible allogeneic hematopoietic stem cell transplantation (allo-HCT) is a common practice and represents a challenging transfusion scenario. Prolonged thrombocytopenia with increased platelet transfusion needs is one of its reported adverse effects, and this has been linked to the persistence of recipient anti-donor isoagglutinins. Case Presentation: A 55-year-old male patient, O Rh(D)-positive, with chronic myelomonocytic leukemia underwent major incompatible allo-HCT from a A Rh(D)-negative donor. He presented with prolonged thrombocytopenia and multiple transfusion reactions after A Rh(D)-negative platelet transfusions. Considering the outcomes of numerous examinations, we tested the anti-A1 titers, finding a significant persistence of anti-donor isoagglutinins. We limited platelet transfusions to blood group O Rh(D)-negative donors, which significantly decreased the requirement for platelet transfusions. In addition, the transfusion reactions ceased. Conclusion: In case of transfusion reactions against platelet products in major ABO-incompatible allo-HCT patients, isoagglutinin monitoring should be considered and a change in the platelet transfusion protocol may be beneficial in patients presenting high isotiters against recipient’s blood type.
导言：ABO不相容异基因造血干细胞移植（allo-HCT）是一种常见的治疗方法，也是一种具有挑战性的输血方案。据报道，血小板减少时间延长、血小板输注需求增加是其不良反应之一，这与受体抗供体异凝集素的持续存在有关。病例介绍：一名 55 岁的男性患者，O 型 Rh（D）阳性，患有慢性粒单核细胞白血病，接受了来自 A 型 Rh（D）阴性供体的主要不相容异体造血干细胞移植。在输注 A 型 Rh(D) 阴性血小板后，他出现了长时间血小板减少和多次输血反应。考虑到多次检查的结果，我们检测了抗 A1 滴度，发现抗供体异凝集素显著持续存在。我们将血小板输注限制在血型为 O 型 Rh（D）阴性的献血者身上，这大大降低了血小板输注的需求量。此外，输血反应也停止了。结论如果主要 ABO 血型不相容的异体肝移植患者出现血小板产品输血反应，应考虑进行等凝集素监测，改变血小板输注方案可能对出现受体血型高等凝集素的患者有益。
{"title":"Prolonged Thrombocytopenia and Severe Transfusion Reaction after ABO-Incompatible Allogeneic Hematopoietic Stem Cell Transplantation in a Patient with Chronic Myelomonocytic Leukemia","authors":"Lina S. Silva-Bermúdez, D. Heidenreich, Stefan A. Klein, Patrick Wuchter, Harald Klüter, Sabine Kayser","doi":"10.1159/000534272","DOIUrl":"https://doi.org/10.1159/000534272","url":null,"abstract":"Introduction: Major ABO-incompatible allogeneic hematopoietic stem cell transplantation (allo-HCT) is a common practice and represents a challenging transfusion scenario. Prolonged thrombocytopenia with increased platelet transfusion needs is one of its reported adverse effects, and this has been linked to the persistence of recipient anti-donor isoagglutinins. Case Presentation: A 55-year-old male patient, O Rh(D)-positive, with chronic myelomonocytic leukemia underwent major incompatible allo-HCT from a A Rh(D)-negative donor. He presented with prolonged thrombocytopenia and multiple transfusion reactions after A Rh(D)-negative platelet transfusions. Considering the outcomes of numerous examinations, we tested the anti-A1 titers, finding a significant persistence of anti-donor isoagglutinins. We limited platelet transfusions to blood group O Rh(D)-negative donors, which significantly decreased the requirement for platelet transfusions. In addition, the transfusion reactions ceased. Conclusion: In case of transfusion reactions against platelet products in major ABO-incompatible allo-HCT patients, isoagglutinin monitoring should be considered and a change in the platelet transfusion protocol may be beneficial in patients presenting high isotiters against recipient’s blood type.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139442847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
MD – Peter Schlenke, PhD – Peter Bugert, MD Beate Mayer, MD Axel Pruß, MD Franz F. Wagner, MD Patrick Wuchter, PhD – Jason Acker, MD Gregor Bein, MD – Reinhard Burger, Robert Koch, PhD Toni Cathomen, PhD Jens Dreier, MD Hermann Eichler, MD – Andreas Humpe, MD Harald Klüter, MD – Jens Meier, MD – Rainer Moog, German Red, PhD – Bristol Andrew D. Mumford, CardioVascular, PhD – Thierry Peyrard, MD – Erwin Strasser, PhD – Pieter F. van der Meer, MD – Mark H. Yazer, E. Strasser, J. Piñeyroa, J. Cid, M. Lozano, P. Schlenke, von Heymann, Berlin Lier, H. Cologne, C. Rosenthal, L. Kaufner, Berlin, P.F.W. Strengers, Amsterdam, J. Cottrell, A. Al Sanani, I. Ogu, D. Chaffin, WV Huntington, D’Alessandro, P. A. Bugert, Research Articles, Meta-Analysis Qin, X. G. Han
{"title":"Contents Vol. 50, 2023","authors":"MD – Peter Schlenke, PhD – Peter Bugert, MD Beate Mayer, MD Axel Pruß, MD Franz F. Wagner, MD Patrick Wuchter, PhD – Jason Acker, MD Gregor Bein, MD – Reinhard Burger, Robert Koch, PhD Toni Cathomen, PhD Jens Dreier, MD Hermann Eichler, MD – Andreas Humpe, MD Harald Klüter, MD – Jens Meier, MD – Rainer Moog, German Red, PhD – Bristol Andrew D. Mumford, CardioVascular, PhD – Thierry Peyrard, MD – Erwin Strasser, PhD – Pieter F. van der Meer, MD – Mark H. Yazer, E. Strasser, J. Piñeyroa, J. Cid, M. Lozano, P. Schlenke, von Heymann, Berlin Lier, H. Cologne, C. Rosenthal, L. Kaufner, Berlin, P.F.W. Strengers, Amsterdam, J. Cottrell, A. Al Sanani, I. Ogu, D. Chaffin, WV Huntington, D’Alessandro, P. A. Bugert, Research Articles, Meta-Analysis Qin, X. G. Han","doi":"10.1159/000535411","DOIUrl":"https://doi.org/10.1159/000535411","url":null,"abstract":"","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139012756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Thomas Frietsch, Maria B. Rondinelli, Jerrold H. Levy
{"title":"Congratulation, Appraisal, and Comment on the 25 Years Anniversary of Serious Hazards of Blood Transfusion","authors":"Thomas Frietsch, Maria B. Rondinelli, Jerrold H. Levy","doi":"10.1159/000532049","DOIUrl":"https://doi.org/10.1159/000532049","url":null,"abstract":"","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139233540","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: Transfusion of platelets is a life-saving medical strategy used worldwide to treat patients with thrombocytopenia as well as platelet function disorders. Summary: Until the end of 1960s, platelets were stored in the cold because of their superior hemostatic functionality. Cold storage of platelets was then abandoned due to better posttransfusion recovery and survival of room temperature (RT)-stored platelets, demonstrated by radioactive labeling studies. Based on these findings, RT became the standard condition to store platelets for clinical applications. Evidence shows that RT storage increases the risk of septic transfusion reactions associated with bacterial contamination. Therefore, the storage time is currently limited to 4–7 days, according to the national guidelines, causing a constant challenge to cover the clinical request. Despite the enormous efforts made to optimize storage conditions of platelets, the quality and efficacy of platelets still decrease during the short storage time at RT. In this context, during the last years, cold storage has seen a renaissance due to the better hemostatic functionality, reduced risk of bacterial contamination, and potentially longer storage time. Key Messages: In this review, we will focus on the impact of cold storage on the in vitro platelet functions as promising alternative storage temperature for future medical applications.
{"title":"In vitro Hemostatic Functions of Cold-Stored Platelets","authors":"J. Kirschall, G. Uzun, T. Bakchoul, I. Marini","doi":"10.1159/000533735","DOIUrl":"https://doi.org/10.1159/000533735","url":null,"abstract":"Background: Transfusion of platelets is a life-saving medical strategy used worldwide to treat patients with thrombocytopenia as well as platelet function disorders. Summary: Until the end of 1960s, platelets were stored in the cold because of their superior hemostatic functionality. Cold storage of platelets was then abandoned due to better posttransfusion recovery and survival of room temperature (RT)-stored platelets, demonstrated by radioactive labeling studies. Based on these findings, RT became the standard condition to store platelets for clinical applications. Evidence shows that RT storage increases the risk of septic transfusion reactions associated with bacterial contamination. Therefore, the storage time is currently limited to 4–7 days, according to the national guidelines, causing a constant challenge to cover the clinical request. Despite the enormous efforts made to optimize storage conditions of platelets, the quality and efficacy of platelets still decrease during the short storage time at RT. In this context, during the last years, cold storage has seen a renaissance due to the better hemostatic functionality, reduced risk of bacterial contamination, and potentially longer storage time. Key Messages: In this review, we will focus on the impact of cold storage on the in vitro platelet functions as promising alternative storage temperature for future medical applications.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139267885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jiaxuan Yang, Aijing Li, Minghao Li, Shulin Ruan, Luyi Ye
Introduction: The Vel– phenotype is a rare blood group, and it is challenging for identifying this phenotype due to limited available reagents. Moreover, there are relatively few studies on genomic editing of erythroid antigens and generation of knockout (KO) cell lines at present. Methods: To identify the high-efficiency small-guiding RNA (sgRNA) sequence, candidate sgRNAs were transfected into HEK 293T cells and analyzed using Sanger sequencing. Following this, the high-efficiency sgRNA was transfected into K562 cells using lentivirus transduction to generate KO Vel blood group gene cells. The expression of the Vel protein was detected using Western blot on single-cell clones. Additionally, flow cytometry was used to detect the erythroid markers CD235a and CD71. Hemoglobin quantification and Giemsa staining were also performed to evaluate the erythroid differentiation of KO clones induced by hemin. Results: The high-efficiency sgRNA was successfully obtained and used for CRISPR-Cas9 editing in K562 cells. After limiting dilution and screening, two KO clones had either deleted 2 or 4 bases and showed no expression of the Vel protein. In the hemin-induced KO clone, there was a significant difference in erythroid marker and hemoglobin quantification compared to untreated cells. The morphological changes were also observed for the hemin-induced KO clone. Conclusion: In this study, a highly efficient sgRNA was screened out and used to generate Vel erythroid antigen KO single-cell clones in K562 cells. The edited cells could then be induced to undergo erythroid differentiation with the use of hemin.
{"title":"CRISPR/Cas9-Editing K562 Cell Line as a Potential Tool in Transfusion Applications: Knockout of Vel Antigen Gene","authors":"Jiaxuan Yang, Aijing Li, Minghao Li, Shulin Ruan, Luyi Ye","doi":"10.1159/000534012","DOIUrl":"https://doi.org/10.1159/000534012","url":null,"abstract":"<b><i>Introduction:</i></b> The Vel– phenotype is a rare blood group, and it is challenging for identifying this phenotype due to limited available reagents. Moreover, there are relatively few studies on genomic editing of erythroid antigens and generation of knockout (KO) cell lines at present. <b><i>Methods:</i></b> To identify the high-efficiency small-guiding RNA (sgRNA) sequence, candidate sgRNAs were transfected into HEK 293T cells and analyzed using Sanger sequencing. Following this, the high-efficiency sgRNA was transfected into K562 cells using lentivirus transduction to generate KO Vel blood group gene cells. The expression of the Vel protein was detected using Western blot on single-cell clones. Additionally, flow cytometry was used to detect the erythroid markers CD235a and CD71. Hemoglobin quantification and Giemsa staining were also performed to evaluate the erythroid differentiation of KO clones induced by hemin. <b><i>Results:</i></b> The high-efficiency sgRNA was successfully obtained and used for CRISPR-Cas9 editing in K562 cells. After limiting dilution and screening, two KO clones had either deleted 2 or 4 bases and showed no expression of the Vel protein. In the hemin-induced KO clone, there was a significant difference in erythroid marker and hemoglobin quantification compared to untreated cells. The morphological changes were also observed for the hemin-induced KO clone. <b><i>Conclusion:</i></b> In this study, a highly efficient sgRNA was screened out and used to generate Vel erythroid antigen KO single-cell clones in K562 cells. The edited cells could then be induced to undergo erythroid differentiation with the use of hemin.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135876462","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Patrick Meybohm, Lotta Hof, Suma Choorapoikayil, Kai Zacharowski
{"title":"Patient Blood Management: We Still Have Work to Do","authors":"Patrick Meybohm, Lotta Hof, Suma Choorapoikayil, Kai Zacharowski","doi":"10.1159/000534087","DOIUrl":"https://doi.org/10.1159/000534087","url":null,"abstract":"","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134973216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Eungjun Yoon, Tae Yeul Kim, Hyungsuk Kim, Duck Cho
Introduction: Evorpacept is a CD47-blocking agent currently being developed for the treatment of various cancers. Interference by evorpacept in pretransfusion compatibility testing has been reported at limited plasma concentrations. Although various mitigation strategies have been proposed, none are practical. This in vitro study assessed evorpacept-induced interference at extended concentrations and investigated the capability of a novel mitigation agent, Evo-NR. Methods: Antibody screening tests were performed on evorpacept-spiked plasma with (anti-E and anti-Jka) or without alloantibodies at evorpacept concentrations up to 2,000 μg/mL using manual gel cards and automated analyzers. Evorpacept-coated red blood cells (RBCs) (rr [ce/ce], Fy[a+b−], S−s+) were tested by direct antiglobulin testing (DAT) and antigen typing using anti-Fyb and anti-S reagents at indirect antiglobulin testing (IAT) phase. Evo-NR was used to resolve the interference in plasma and RBC samples. Flow cytometry was used to assess the mitigation effects. Results: Evorpacept-spiked plasma showed panreactive interference in antibody screening tests using manual gel cards (2+ to 3+) and automated analyzers (4+). A carryover effect was also observed in the automated analyzers. The use of a 3- to 6-fold molar excess of Evo-NR effectively resolved the interference in the plasma and enabled accurate alloantibody identification. Although the reduction in evorpacept binding to RBCs was identified via flow cytometry, Evo-NR was incapable of resolving the serologic interference observed in DAT and antigen typing at IAT phase. Discussion: Evorpacept showed constant panreactivity and a carryover effect at high concentrations. Evo-NR successfully resolved the interference in the plasma samples and could be considered a practical and efficient mitigation solution. Implementation of Evo-NR has the potential to support RBC transfusion for patients undergoing evorpacept treatment.
{"title":"Evorpacept-Induced Interference and Application of a Novel Mitigation Agent, Evo-NR, in Pretransfusion Testing","authors":"Eungjun Yoon, Tae Yeul Kim, Hyungsuk Kim, Duck Cho","doi":"10.1159/000534273","DOIUrl":"https://doi.org/10.1159/000534273","url":null,"abstract":"<b><i>Introduction:</i></b> Evorpacept is a CD47-blocking agent currently being developed for the treatment of various cancers. Interference by evorpacept in pretransfusion compatibility testing has been reported at limited plasma concentrations. Although various mitigation strategies have been proposed, none are practical. This in vitro study assessed evorpacept-induced interference at extended concentrations and investigated the capability of a novel mitigation agent, Evo-NR. <b><i>Methods:</i></b> Antibody screening tests were performed on evorpacept-spiked plasma with (anti-E and anti-Jk<sup>a</sup>) or without alloantibodies at evorpacept concentrations up to 2,000 μg/mL using manual gel cards and automated analyzers. Evorpacept-coated red blood cells (RBCs) (rr [ce/ce], Fy[a+b−], S−s+) were tested by direct antiglobulin testing (DAT) and antigen typing using anti-Fy<sup>b</sup> and anti-S reagents at indirect antiglobulin testing (IAT) phase. Evo-NR was used to resolve the interference in plasma and RBC samples. Flow cytometry was used to assess the mitigation effects. <b><i>Results:</i></b> Evorpacept-spiked plasma showed panreactive interference in antibody screening tests using manual gel cards (2+ to 3+) and automated analyzers (4+). A carryover effect was also observed in the automated analyzers. The use of a 3- to 6-fold molar excess of Evo-NR effectively resolved the interference in the plasma and enabled accurate alloantibody identification. Although the reduction in evorpacept binding to RBCs was identified via flow cytometry, Evo-NR was incapable of resolving the serologic interference observed in DAT and antigen typing at IAT phase. <b><i>Discussion:</i></b> Evorpacept showed constant panreactivity and a carryover effect at high concentrations. Evo-NR successfully resolved the interference in the plasma samples and could be considered a practical and efficient mitigation solution. Implementation of Evo-NR has the potential to support RBC transfusion for patients undergoing evorpacept treatment.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135219224","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Johannes-Moritz von Behren, Jan Wesche, Andreas Greinacher, Konstanze Aurich
Introduction: Before being implemented in daily clinical routine, new production strategies for platelet concentrates (PCs) must be validated for their efficacy. Besides in vitro testing, the establishment of new methods requires the labeling of platelets for in vivo studies of platelets’ survival and recovery. Indocyanine green (ICG) is a Food and Drug Administration-approved near-infrared (NIR) fluorescent dye for diagnostic use in vivo, suitable for non-radioactive direct cell labeling of platelets. Methods: Platelets from PCs in storage solutions with different plasma concentrations were labeled with ICG up to concentrations of 200 μm. Whole blood (WB) was used as an ex vivo matrix to monitor the labeling stability of ICG-labeled platelets. The impact of labeling processes was assessed by the quantification of CD62P expression and PAC-1 binding as platelet function markers. Platelet aggregation was analyzed by light transmission aggregometry. ICG-labeling efficiency and stability of platelets were determined by flow cytometry. Results: Platelets from PCs could be successfully labeled with 10 μm ICG after 1 and 4 days of storage. The best labeling efficiency of 99.8% ± 0.1% (immediately after labeling) and 81% ± 6.2% (after 24 h incubation with WB) was achieved by plasma replacement by 100% platelet additive solution for the labeling process. Since the washing process slightly impaired platelet function, ICG labeling itself did not affect platelets. Immediately after the ICG-labeling process, plasma was re-added, resulting in a recovered platelet function. Conclusion: We developed a Good Manufacturing Practice compatible protocol for ICG fluorescent platelet labeling suitable for survival and recovery studies in vivo as a non-radioactive labeling alternative.
{"title":"Indocyanine Green-Labeled Platelets for Survival and Recovery Studies","authors":"Johannes-Moritz von Behren, Jan Wesche, Andreas Greinacher, Konstanze Aurich","doi":"10.1159/000533623","DOIUrl":"https://doi.org/10.1159/000533623","url":null,"abstract":"<b><i>Introduction:</i></b> Before being implemented in daily clinical routine, new production strategies for platelet concentrates (PCs) must be validated for their efficacy. Besides in vitro testing, the establishment of new methods requires the labeling of platelets for in vivo studies of platelets’ survival and recovery. Indocyanine green (ICG) is a Food and Drug Administration-approved near-infrared (NIR) fluorescent dye for diagnostic use in vivo, suitable for non-radioactive direct cell labeling of platelets. <b><i>Methods:</i></b> Platelets from PCs in storage solutions with different plasma concentrations were labeled with ICG up to concentrations of 200 μ<sc>m</sc>. Whole blood (WB) was used as an ex vivo matrix to monitor the labeling stability of ICG-labeled platelets. The impact of labeling processes was assessed by the quantification of CD62P expression and PAC-1 binding as platelet function markers. Platelet aggregation was analyzed by light transmission aggregometry. ICG-labeling efficiency and stability of platelets were determined by flow cytometry. <b><i>Results:</i></b> Platelets from PCs could be successfully labeled with 10 μ<sc>m</sc> ICG after 1 and 4 days of storage. The best labeling efficiency of 99.8% ± 0.1% (immediately after labeling) and 81% ± 6.2% (after 24 h incubation with WB) was achieved by plasma replacement by 100% platelet additive solution for the labeling process. Since the washing process slightly impaired platelet function, ICG labeling itself did not affect platelets. Immediately after the ICG-labeling process, plasma was re-added, resulting in a recovered platelet function. <b><i>Conclusion:</i></b> We developed a Good Manufacturing Practice compatible protocol for ICG fluorescent platelet labeling suitable for survival and recovery studies in vivo as a non-radioactive labeling alternative.","PeriodicalId":23252,"journal":{"name":"Transfusion Medicine and Hemotherapy","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136078834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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