Random Donor Research Articles

Two Concurrent Cases of NAIT in Pregnant Sisters

Abstract Neonatal alloimmune thrombocytopenia (NAIT) is caused by maternal antibodies directed against fetal platelet antigens inherited from the father. Approximately 80% of cases of NAIT are attributed to antibodies against human platelet antigen (HPA)-1a and less commonly to antibodies against HPA-5b. We present two cases of possible NAIT attributed to anti-HPA-5b that were identified in sisters requiring concurrent management. Patient #1 is a 32-year-old G9P4044 woman with one pregnancy complicated by intrauterine growth retardation and partial placental abruption, another pregnancy complicated by fetal anomaly (intracerebral hemorrhage) status post termination of pregnancy at 23 weeks’ gestation and three miscarriages (one with chromosomal abnormality). The patient is homozygous for HPA-1a and HPA-5a, and the partner is homozygous for HPA-5b. HPA-5b antibodies and HLA antibodies were identified in the patient’s serum. Deemed at high risk for subsequent NAIT, she was started on intravenous immunoglobulin twice weekly and steroids during her second trimester until delivery. The patient underwent weekly fetal sonograms during pregnancy which were normal. The patient’s platelet counts were stable. The patient refused caesarean delivery. At 38 weeks’ gestation, the patient gave birth to a healthy female via normal spontaneous vaginal delivery. HPA-5b negative platelets were procured from our blood supplier, but neither the mother nor neonate required transfusion. The neonate showed no signs of bleeding and had normal platelet counts. Ultrasound imaging of the head was unremarkable. The patient and neonate were discharged home. Patient #2 is a 33-year-old G13P6156 woman with a prior pregnancy complicated by stillbirth at 32 weeks’ gestation. Considering her personal and family history, HPA testing was performed and HPA-5b antibodies were identified in the patient’s serum. The patient’s partner is heterozygous for HPA-5b. Crossmatched platelets were procured from our blood supplier. The patient refused caesarean delivery. The patient gave birth at 37 weeks’ gestation to a healthy male via normal spontaneous vaginal delivery. The fetus had mild thrombocytopenia (107 x 103/uL), with no bleeding or bruising identified. Neither the mother nor neonate required transfusion. The patient and neonate were discharged home. Ultrasound imaging of the head was unremarkable, and his thrombocytopenia resolved. While some studies postulate that anti-HPA-5b identification may be coincidental rather contributory to the disease process, these cases re-emphasize the importance of obtaining pregnancy, neonatal and family history to best risk stratify patients and determine management from both the clinical and transfusion medicine perspectives. If transfusion is needed, transfusion options include HPA negative platelets, crossmatched platelets and random donor platelets depending on clinical urgency and platelet availability.

Maternal Immune Thrombocytopenic Purpura Leading to Severe Neonatal Autoimmune Thrombocytopenia: Report of Two Cases

Introduction: In neonatal intensive care units, neonatal thrombocytopenia is one of the common hematological abnormality seen. Neonatal autoimmune thrombocytopenia should be considered in any neonate who is born to a known case of immune thrombocytopenia purpura (ITP) mother, with early onset thrombocytopenia without any signs of sepsis. Neonatal ITP is a condition of autoantibody mediated platelet destruction. Case details: Two neonates with thrombocytopenia, born to mothers with ITP are described in this report. Lowest platelet count noted was 7000 cells/cmm in one of the neonate. Both neonates received intra venous immunoglobulin (IVIG) while one neonate had persistent and severe thrombocytopenia requiring multiple random donor platelet (RDP) transfusions followed by oral steroid as well. Conclusion: Neonatal thrombocytopenia associated with maternal ITP need close monitoring, early sampling and diagnosis to prevent any possible complications and warrant early initiation of treatment.

Categorization of the Inevitable Causes of Blood and Components Wastage Detected in South Indian Urban Blood Centre

Background: Since there is no artificial substitute for human blood, it plays a vital role in the patient management in this era of modern medicine. Therefore the donated blood from a healthy donor is precious and must not be wasted without proper utilization. Blood wastage is a universal phenomenon which needs a careful approach with strict guidelines. The study aims at categorizing the various inevitable causes of wastage with an attempt to determine ways of preventing the loss due to non-utilization. Materials and Methods: A retrospective – cross sectional study was conducted in a newly started Stand-alone Blood Centre in South Indian Urban city between June, 2022 and June 2024.All the components including Whole Blood (WB), Packed Red Blood Cell Concentrate (PRBC), Platelet concentrate (PC) and Fresh Frozen Plasma (FFP) were included in the study. The reasons for blood and component wastage was noted down from the various Registers of the Blood Centre and all the data was analyzed using Microsoft Excel Spread Sheet. Results: A total of 2456 units of blood were collected. 5284 components were prepared from these which includes 42 units of Whole Blood, 2397 Packed Red Blood cells(PRBC), 2393 Fresh frozen Plasma(FFP), 452 Platelet Concentrate(PC).Overall discard rate was found to be 11.90%. PC discard rate was highest (49.34%) followed by WBU (19.04%), PRBC (13.01%) AND FFP (3.59%). The commonest cause for the discarding was expiry of the shelf life of blood bags (74.72%) followed by damaged bags (9.69%) and TTI’s (6.76%). Conclusion: Blood and its components wastage is an inevitable reality which needs to be approached as upmost priority. Zero wastage of blood and components may be targeted by recommendations like appropriate training of the blood bank staff, keeping negative group donors on reserve, strictly following FIFO policy while dispensing blood, whole blood unit collection may be minimized, encouraging Single Donor Platelets (SDP) by apheresis instead of Random Donor Platelets (RDP) preparation, Blood Utilization Committee to be made mandatory and permission for reciprocal sharing of blood components among blood banks.

A rare case of hemolytic disease-associated thrombocytopenia mimicking alloimmune thrombocytopenia: Salvaging a precious baby

Abstract: Thrombocytopenia associated with hemolytic disease of fetus and newborn (HDFN) is usually mild-to-moderate severity. Neonatal alloimmune thrombocytopenia (NAIT) is one of the leading causes of severe thrombocytopenia in neonates. We describe a rare case of very severe thrombocytopenia (5 g/L) with bleeding manifestations associated with HDFN mimicking NAIT. The baby was a preterm female, born to a 34-year-old multiparous mother with a bad obstetric history and multiple alloantibodies (anti-D, anti-C, anti-E, and anti-Jka). She was managed with phototherapy, exchange transfusion with phenotype-matched packed red blood cells, random donor platelets and intravenous immunoglobulin as platelet crossmatching could not provide compatible platelets. NAIT was ruled out with human platelet antigen (HPA) genotyping of the baby, the mother, and the father. It was concluded that the very severe thrombocytopenia might be due to HDFN after ruling out the other causes. HDFN can rarely cause very severe thrombocytopenia. However, when a newborn presents with severe/very severe thrombocytopenia, NAIT is to be ruled out with HPA genotyping, even in resource-limited settings, to plan out subsequent pregnancies.

Thromboelastography-Guided versus Standard-of-Care or On-Demand Platelet Transfusion in Patients with Cirrhosis and Thrombocytopenia Undergoing Procedures: A Randomized Controlled Trial

PurposeTo determine the rate of platelet transfusion in patients with cirrhosis and severe thrombocytopenia (platelet counts <50 × 109/L) undergoing high-risk invasive procedures when prescribed by thromboelastography (TEG) compared with empirical and on-demand transfusion strategies. Materials and MethodsThis was a single-center, single-blinded, randomized controlled trial. Patients with cirrhosis and severe thrombocytopenia undergoing high-risk invasive procedures were randomized into 3 groups: TEG group, transfusions based on TEG parameters; standard of care (SOC) group, 3 units of random donor platelets before procedure; and on-demand group, transfusions based on procedural adverse events/clinician’s discretion. The primary outcome was periprocedural platelet transfusion in each arm. ResultsEighty-seven patients were randomized (29 in each group) with no significant differences in demographics/coagulation profile/procedures. The median platelet count was 33 × 109/L (interquartile range, 26–43 × 109/L). Percutaneous liver biopsy was the most common procedure (46, 52.9%). Significantly lower number of patients in the TEG group received platelets (4 cases, 13.8%; 95% CI, 3.9–31.7) compared with SOC group (100%; 95% CI, 88.1–100; P < .001). Four patients in the on-demand group received platelets (13.8%; 95% CI, 3.9–31.7). Minor (World Health Organization [WHO] Grade 2) procedure-related bleeding occurred in 3 (10%; 95% CI, 2.2–27.4) patients in the TEG-guided transfusion group compared with 1 (3.4%; 95% CI, 0.1–17.8) patient each in the SOC and on-demand groups (P = .43), although the study was not powered for comparison of bleeding rates. No bleeding-related mortality was observed in any of the 3 groups. ConclusionsTEG-prescribed transfusion reduced prophylactic transfusions in patients with cirrhosis and severe thrombocytopenia undergoing high-risk invasive procedures. The study was not powered for comparison of bleeding rates.

Frequency of Blood Donor Deferrals in Urban Population among Random Voluntary Donors and Evaluation of Gross Differences in Various Study Groups

Introduction: The most important aspect of the blood donor selection criteria is the selection of Healthy donors for donating blood to safeguard the health of the donor as well as the patient. Based on the selection criteria donors are considered either fit or unfit (deferred) to donate blood. Aim: Primary aim of the study is to analyze and evaluate the various causes of donor deferrals in comparison to different regional studies with special emphasis on temporary donor deferrals, in order to retain them as potential future donors. Materials &amp; Methods: A retrospective study was conducted with available data of all donors registered between May 2022 and Dec 2023. Donor deferral data was analyzed for various causes of deferral patterns. Results: A total of 2003 donors registered for voluntary blood donation out of which 244(80.79%) were accepted for donation and 302(15.07 %.) were deferred for various reasons. Total number of temporary deferrals were 244(80.79%) and permanent deferrals were 58(19.20%). Discussion: The overall deferral rate in this study was 15.48%. Low Hemoglobin (36.09%) was the commonest cause for temporary deferral and the commonest age group which had maximum deferrals was 18---25 yrs. Hypertension was the main cause of permanent deferral. Conclusion: The incidence of donor deferral can be reduced by furnishing the appropriate information related to blood donations and educating the general public and blood donors through awareness programs. Follow up of the donors must be encouraged to remove the negative feeling in deferred donors to make them potential future donors.

Chemical and Microbiological Changes of Expired Platelet Concentrate.

Platelets are a commonly used blood component to prevent or treat bleeding in patients with thrombocytopenia or platelet dysfunction. They are stored at room temperature (22-24°C) for five days unless specific measures are taken to extend the shelf life to seven days or more. After five days, this study evaluated platelet units' biochemical changes and bacterial growth. Platelet concentrate was collected from 30 random donors: 8 females and 22 males. The collected samples were then placed on an agitator at room temperature and tested for their pH, protein content, and glucose levels using Roche Combur 100 Test® Strips. The Haemonetics eBDS™ System was used for bacterial detection. The measurements were taken on day five as the control and then repeated on days 7, 9, and 11 to observe any changes. On days 5 and 7, all parameters remained unchanged. However, glucose levels significantly changed (p=<0.0001) on days 9 and 11. Regarding pH, a significant change was observed on day 9 (p=0.033) and day 11 (p=0.0002). There were no significant changes in all parameters on days 5 and 7. However, glucose was substantially changed (p=<0.0001) on days 9 and 11. For pH, there was a significant change in pH on day 9 (p=0.033) and day 11 (p=0.0002). Our study found that platelet concentrate extension is possible for up to seven days. However, further studies are needed to evaluate platelet function during expiry time and to assess the stability of platelet morphology and function.

Evidence that CD36 is expressed on red blood cells and constitutes a novel blood group system of clinical importance.

Polymorphic molecules expressed on the surface of certain blood cells are traditionally categorized as blood groups and human platelet or neutrophil antigens. CD36 is widely considered a platelet antigen (Naka) and anti-CD36 can cause foetal/neonatal alloimmune thrombocytopenia (FNAIT) in CD36-negative pregnant women. CD36 is used as a marker of differentiation in early erythroid culture. During the experimental culture of CD34+ cells from random blood donors, we observed that one individual lacked CD36. We sought to investigate this observation further and determine if CD36 fulfils the International Society of Blood Transfusion criteria for becoming a blood group. Surface markers were monitored by flow cytometry on developing cells during the erythroid culture of CD34+ cells. Genetic and flow cytometric analyses on peripheral blood cells were performed. Proteomic datasets were analysed, and clinical case reports involving anti-CD36 and foetal anaemia were scrutinized. Sequencing of CD36-cDNA identified homozygosity for c.1133G>T/p.Gly378Val in the CD36-negative donor. The minor allele frequency of rs146027667:T is 0.1% globally and results in abolished CD36 expression. CD36 has been considered absent from mature red blood cells (RBCs); however, we detected CD36 expression on RBCs and reticulocytes from 20 blood donors. By mining reticulocyte and RBC datasets, we found evidence for CD36-derived peptides enriched in the membrane fractions. Finally, our literature review revealed severe cases of foetal anaemia attributed to anti-CD36. Based on these findings, we conclude that CD36 fulfils the criteria for becoming a new blood group system and that anti-CD36 is implicated not only in FNAIT but also foetal anaemia.

Fully genotyping and screening of clinically important blood-group antigens by MALDI TOF mass spectrometry.

Several molecular biology methods are available for high-throughput blood typing. In this study, we aimed to build a high-throughput blood-group genetic screening system for high-frequency blood-group antigen-negative rare-blood groups in donors and patients. The amplification primers for all blood-type gene fragments involving the selected alleles were designed for detection. Single-base extend primers were also designed based on specific loci. DNA fragments were detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS) for the last nucleotide identification of amplification products in the extend step. The accuracy was verified by known samples. Thirty-six random samples were detected by serological tests and sequencing to verify the system stability. After verification, according to the collected known rare-blood-type samples, all the alleles designed to be detected matched with the validated single-nucleotide polymorphisms. The verification tests showed that all genotyping results of the random samples were in accordance with the findings of serotyping and sequencing. Then, 1258 random donor samples were screened by the built typing system after the verification. Three Fy(a-) and four s- were screened out in 1258 random blood samples. The multiple polymerase chain reaction-based MS detection system can be used in rare-blood-type screening with good accuracy and stability.

Growth and Distribution of Bacteria in Contaminated Whole Blood and Derived Blood Components

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.

Impact of Premedication Prior to Platelet Transfusion on Platelet Increment Among Hemato-Oncological Patients

Introduction: Platelet transfusions have reduced the incidence of major hemorrhagic complications among hemato-oncological patients, however they are associated with potential complications, including transfusion reactions. The efficacy of premedication with steroids and/or antihistamines prior to platelet transfusion in preventing transfusion reactions has been studied. However, to the best of our knowledge, the effect of premedication on platelet increment has not been previously addressed. We aimed to assess the impact of premedication on platelet increment after platelet transfusion. Methods: We conducted a prospective study among adult patients admitted to the hemato-oncology and bone marrow transplantation (BMT) units in a tertiary medical center. Enrolled patients, who received a platelet transfusion during their admission, had a complete blood count taken 1-hour post platelet transfusion completion. The decision whether to administer premedication and the specific medications were left to the discretion of the attending physician. Single donor platelets or 5 pooled random donor platelets were provided by the blood bank interchangeably according to inventory level. The primary outcome was platelet recovery, defined as the difference between the 1-hour post- and pre- transfusion platelet counts. Adequate platelet recovery was considered &amp;gt;10,000/µL. Secondary outcome was platelet survival, defined as the difference between platelet count 18- to 24-hours post- and pre- transfusion. Rates of post-transfusion reactions and premedication complications were recorded. Association of other clinical variables with outcomes of interest was evaluated and variables significantly associated with adequate platelet recovery response (p&amp;lt;0.05) were entered into a multivariable analysis. Results: Overall, 68 patients were enrolled, with a median age of 58 years (interquartile range [IQR] 45-67) and 43 (63%) were males. The most common diagnoses were AML (n=24, 35%) and multiple myeloma (n=23, 34%). Most patients (72%) were admitted for a BMT (autologous 47%, allogeneic 25%). Outcomes were recorded in response to 217 platelet transfusions, with a median number of platelet transfusion per patient of 4 (IQR 3-5). Premedication was given prior to 64 transfusions (29%), consisting of steroids in 17 (27%), antihistamines in 4 (6%), or both in 43 (67%) platelet transfusions. In 153 (71%) platelet transfusions, no premedication was given. Patients who received premedication were younger, more often had a diagnosis of acute leukemia and were mostly admitted for non-transplant admissions. ( Table 1). Median platelet recovery and survival for all platelet transfusions were 15,000/µL (IQR 10,0000-22,500/µL) and 8,000/µL (IQR 4000-14,000/µL), respectively. There was no significant difference in median platelet recovery and survival between patients who received premedication and those who did not (15,000/µL in both cohorts, p=0.36 and 6,500 vs. 9,000/µL, p=0.69, respectively). The number of platelet transfusions leading to adequate platelet recovery was 150/213 (70.4%) overall, with no significant difference between patients who received premedication and those who did not [45/64 (70.3%) vs.105/153 (70.5%), p=1.0]. Female gender was associated with a higher rate of adequate platelet response in univariate analysis (81.7% vs. 63.4%, p=0.006), while splenomegaly was associated with a trend towards lower rates of adequate platelet recovery (55.2% vs. 71.4%, p=0.086). None of these variables was associated with adequate platelet recovery in multivariate analysis. Febrile non-hemolytic transfusion reaction (FNHTR) was significantly more common after platelet transfusions without premedication compared to platelet transfusions with premedication (7.8% vs. 0%, respectively, p=0.02). Among patients who received premedication, only one patient developed grade 2 asymptomatic hyperglycemia. Conclusions: To the best of our knowledge, this is the first prospective study to assess the impact of premedication prior to platelet transfusion on platelet recovery and survival. We did not observe an effect of premedication on various parameters of platelet increment, while use of premedication did reduce FNHTR. Our results suggest that premedication with steroids and/or antihistamines can be given prior to platelet transfusion without compromising platelet increment.

Evaluation of Blood Transfusion-Related Reactions of Blood Recipients in Hamadan Besat Hospital Patients during 2020- 2022

Background: Blood transfusion has some complications known as transfusion-related reactions. Accurate identification and recording of these reactions is more important for their management and prevention. This study aimed to evaluate blood transfusion-related reactions in blood recipients in patients in Hamadan Besat hospital during 2020-2022. Methods: In this cross-sectional study, the data of 400 patients who had complications after receiving blood and blood components were collected in terms of hemolytic and non-hemolytic blood transfusion reactions. Data were collected from patients’ medical records and standard blood transfusion checklists and analyzed by SPSS version 26 software using the chi-square test and Fisher’s exact test. Results: Red blood cell (RBC), 70.25%, random donor platelet (RDP), 10.3%, and fresh frozen plasma (FFP), 9.5%, were the most transfused blood components. Blood transfusion reactions were allergic reactions (53.5%), febrile non-hemolytic transfusion reaction (FNHTR) (24%), dyspnea (10.25%), hemolytic reactions (HR) (4.5%), hypotension (4.25%), transfusion-associated cardiovascular overload (TACO) (1.75%), and other reactions (2.75%). A significant relationship was also observed between receiving components containing RBC, including whole blood (WB0), RBC concentrate, and leukoreduced RBC, and the incidence of FNHTR (P=0.001). Conclusion: Based on the findings of the present study, allergic reaction and FNHTR were the most common complications after blood transfusions. Blood transfusion-related reactions could be predicted, managed, and possibly prevented by considering the relationship between the type of reaction and the type of transfused blood components.

Phase I non-randomized clinical trial of allogeneic natural killer cells infusion in acute myeloid leukemia patients

IntroductionA new type of immune cell transplantation called allogeneic NK cell infusion is proposed as a potential universal off-the-shelf cell product for adoptive immune cell therapy in hematologic malignancies.DesignA multicentral phase I non-randomized clinical trial was conducted to assess the safety, feasibility, and potential efficacy of adoptively infused NK cells in patients with refractory/relapsed AML. We evaluated the feasibility of the trial by considering cell production, patient selection, and treatment protocol.MethodAllogeneic NK cells were produced from random healthy unrelated donors; 10 patients were selected according to the inclusion criteria and were included in two groups in case of NK cell dose escalation. Two cell infusions were given, spaced 7 days apart, following a lymphodepletion conditioning regimen of fludarabin-endoxan administered 7 days before the first infusion. The intervention safety was scored using Common Terminology Criteria for Adverse Events (CTCAE) based on variations in vital signs due to cell infusion. NK cell chimerism, tumor burden, and duration of relapse were considered to be components of efficacy. The pilot feasibility evaluation was checked using the CONSORT platform.ResultsThe NK cell infusion procedure was well tolerated, and no grade 2–5 toxicities related (possible or probable) to PB-NK cell infusion were observed. Four patients developed grade 1 transient chills, headaches, vomiting, and bone pain following each PB-NK cell infusion that were not required hospitalization. One of these patients (p01) died because of severe acute respiratory syndrome. Of 9 evaluable patients, 6 (66.6%) showed stable disease (SD) and 3 (33.3%) presented progressive disease (PD). Of 6 SD patients, 2 (p08 and p09) remained alive in SD and 3 patients (p04, p05 and p07) converted to PD at 9 months after infusion of NK cells, and 1 (p03) was not evaluable due to follow-up loss. No patient achieved complete remission.ConclusionThe study demonstrated the feasibility and safety of adoptive transfer of random healthy unrelated donor PB-NK cells in refractory/relapsed AML patients and supports continued study in phase II clinical trials in relapsed/refractory AML patients.

An Evaluation of Random Donor Platelets Prepared from Overnight Stored Buffy Coat Method: A Better Alternative to the 6-h Restriction.

Preparing maximum platelets from all daily blood donations is essential to maintain adequate stock of platelets that may be feasible by storing buffy coat overnight. The overnight stored buffy coat method is prevalent in Europe and was not studied much in India. Therefore, a comparative study was planned to compare RDPs prepared from fresh and overnight stored buffy coat methods.In this study total of eighty RDPs were prepared. Forty RDPs were prepared by each method by matching similar platelet counts in whole blood in pairs and avoiding other bias. Quality analysis was done in various hematological, biochemical, metabolic, and activation parameters and with National guideline.RDPs prepared by the overnight stored buffy coat method had statistically higher mean platelet counts, lesser WBC contamination, lesser glucose concentration, higher lactate concentration, lesser pH, and higher MPV & PDW than RDPs prepared by the fresh buffy coat method. P selectin levels were lesser in the overnight stored category. The platelet yield in RDPs (extraction from the whole blood) was more in the overnight stored category. Compared with the national guideline, 87.5% of units passed the quality criteria in the fresh category compared to 100% of units in the overnight category.RDPs prepared from overnight stored buffy coats not only had better quality parameters but also were in a less activated state. Because of better quality and feasibility in blood centers, the overnight stored buffy coat method may also be used along with the traditional preparation method.

Cytomegalovirus-Seronegative Plus Leukoreduced Blood Products Are Still Superior to Only Leukoreduced Products in Preventing Transfusion-Transmitted Cytomegalovirus Infection

Introduction Cytomegalovirus (CMV) is one of the most important pathogens causing complications after hematopoietic stem cell transplantation (HSCT). Transfusion-transmitted CMV infection (TT-CMV) in CMV-seronegative patients is a major issue. Leukoreduced (LR) blood products were unavailable before 2004. However, they have gradually become widespread, and all blood products in Japan have been LR since 2007. This study aimed to assess the incidence of TT-CMV in CMV-seronegative HSCT recipients undergoing autologous HSCT (auto-HSCT) or allogeneic HSCT (allo-HSCT) from CMV-seronegative stem cell donors, who received transfusions with “CMV-seronegative products” from CMV-seronegative donors or “random donor products” from donors not tested for anti-CMV antibodies during the periods of availability and unavailability of LR products. Methods We retrospectively evaluated 3590 CMV-seronegative recipients registered in the TRUMP database of the Japanese Society for Transplantation and Cellular Therapy from 1999 to 2018. The recipients undergoing HSCT were categorized into three periods: Period A (when LR products were unavailable), 1999-2003 (n = 136); Period B (when there were both non-LR and LR products), 2004-2006 (n = 200); and Period C (when only LR products were available), 2007-2018 (n = 3254). TT-CMV was defined by the presence of CMV antigenemia and/or CMV diseases. This study was approved by the Ethics Committee of Kobe University Hospital and was conducted in accordance with the principles of the Declaration of Helsinki. Results The median age of the patients at HSCT was 40 years (range, 0-79 years). Among auto-HSCT patients (n = 1628) transfused with “CMV-negative products”(CN group) and “random donor products” (RD group), the incidences of TT-CMV in the CN and RD groups were 0% (0/4) and 3.1% (1/32) during Period A (P = 1.0), 22.2% (2/9) and 10.6% (7/66) during Period B (P = 0.29), and 3.8% (4/105) and 5.0% (71/1412) during Period C (P = 0.81), respectively. Among the allo-HSCT patients (n = 1962), the incidences of TT-CMV in the CN and RD groups were 10.0% (3/30) and 27.1% (19/70) during Period A (P = 0.07), 17.9% (5/28) and 29.9% (29/97) during Period B (P = 0.24), and 11.6% (41/355) and 28.0% (387/1382) during Period C (P &amp;lt; 0.001), respectively. During Period C, 99 patients in the CN group always underwent transfusion with leukocyte reduction filters, of which 11 (11.1%) were diagnosed with TT-CMV. In contrast, 202 patients in the CN group always underwent transfusion without the filters, of which 26 (12.9%) were diagnosed with TT-CMV (P = 0.78). In the RD group, 494 patients always underwent transfusion with leukocyte reduction filters, of which 124 (25.1%) were diagnosed with TT-CMV. On the other hand, 807 patients in the RD group always underwent transfusion without the filters, of which 245 (30.4%) were diagnosed with TT-CMV (P = 0.10). Multivariate analysis revealed that random donor products (hazard ratio [HR]: 3.11, 95% confidence interval [CI]: 2.13-4.52; P &amp;lt; 0.001) and age ≥16 years (HR: 2.90, 95% CI: 2.06-4.09; P &amp;lt; 0.001) were associated with an increased risk of TT-CMV in allo-HSCT. No significant difference was observed in the incidence of CMV diseases between the CN group (3.7%; 13/355) and the RD group (3.8%; 52/1382) (P = 1.00). The major CMV diseases included CMV colitis (n = 5) and pneumonia (n = 2) in the CN group, and CMV colitis (n = 30), pneumonia (n = 9), and retinitis (n = 4) in the RD group. Interestingly, among the patients with TT-CMV during Period C, 27/41 patients (65.9%) in the CN group and 333/387 patients (86.1%) in the RD group received early intervention with anti-CMV agents (P = 0.002). The limitation included that complement-fixation test for anti-CMV antibody was used in Japan Marrow Donor Program in this cohort. Conclusion This is the largest study demonstrating that CMV-seronegative plus LR products are still superior to only LR products from random donors in terms of TT-CMV prevention among allo-HSCT CMV-seronegative recipients. However, no significant difference was observed in the incidence of CMV diseases, possibly because of early intervention with anti-CMV agents.

Impact of Non-Immune Factors on Platelet Transfusion Responses in an Allo-Immunized Cohort

Background: Patients with hematologic malignancies and/or hematopoietic stem cell transplantation (HSCT) recipients often require platelet transfusion support for extended periods. This can result in platelet transfusion refractoriness (PTR) from HLA alloimmunization in up to 30% of individuals. These medically complex patients may also have splenomegaly, fevers and other factors which contribute to PTR. Consequently, they may demonstrate variability in immediate platelet recovery (at 0-2 hours post-transfusion), and in subsequent platelet consumption (at 18-24 hours) despite receiving HLA-selected platelet products. We sought to examine patient and product related factors which impacted platelet transfusion response in a cohort of HLA-alloimmunized platelet transfusion refractory patients. Methods: From April 2021 to June 2023, HLA-alloimmunized (PRA &amp;gt;50%) subjects who required three or more HLA selected platelet units were identified at the Fred Hutchinson Cancer Center/University of Washington. Data were obtained on patient demographics, diagnoses, Panel Reactive Antibodies (PRA %) and factors shown to be associated with poor platelet responses (spleen size, disseminated intravascular coagulation (DIC), bleeding, selected medications, history of hematopoietic stem cell transplant (HSCT) and/or graft versus host disease (GVHD), ABO compatibility, and product age). Platelet corrected count increments (CCI) were calculated for each transfusion. Non-HLA factors potentially contributing to CCIs at 0-2 and 18-24 hours post-transfusion were compared among HLA-compatible (either 4/4 HLA matched without HLA antigen mismatches, or with permissive mismatches (defined as cumulative a mean fluorescent intensity (MFI) &amp;lt;10,000 on the Luminex donor specific HLA antibody testing platform)) and HLA incompatible transfusions (cumulative MFI &amp;gt;10,000 or random donor apheresis products). Results: Of the 420 patients screened, 75 had sufficient records for inclusion. These subjects received 2729 platelet transfusions, an average of 36 transfusions per patient over 5.2 (± 4.1) months. Post transfusion platelet counts were available for 1528 transfusions. All platelet products were leukoreduced, irradiated, single donor apheresis products. Patients were predominantly Caucasian (70.7%; n=53/75), female (82.7%; n=62/75), aged 57 (±14.9) years. The primary diagnosis was Acute myelogenous leukemia or myeloproliferative disorder in most cases (88%; n=66/75). 58.7% (n=44/75) of patients were HSCT recipients (3 auto-; 41 allo-HSCT). Average PRA among patients was 72.4%±22.2%. 25 patients had splenomegaly. Among HLA-compatible transfusions, average CCIs at 0-2 and 18-24 hours were 17,088 and 6,666. In HLA-mismatched/random donor transfusions, average CCIs at 0-2 and 18-24 hours were 11,489 and 2,786; HLA compatible products increased CCIs by 5,599 and 3880 at 0-2 and 18-24 hours. In both groups splenomegaly had the most consistent association with poor CCI at both time intervals (Figures A and B). Conclusions: In this cohort of individuals with HLA-alloimmunization mediated PTR, the administration of HLA-compatible products offset the impact of other detrimental factors generating adequate CCIs at 0-2 and 18-24 hours. Among patients with splenomegaly, CCIs were lower despite HLA compatible product administration. In the same individuals, HLA-incompatible/random donor products negatively impacted CCIs overall and the negative effect of other concomitant non-immune/product-related factors was additive.Our data highlights the need to account for non-HLA related conditions while evaluating CCI response following each HLA-selected product transfusion, and to further explore any interactions among such factors. Next steps include conducting a multivariate analysis and developing a predictive model for platelet transfusion response in HLA-alloimmunized recipients who have concomitant non-immune or product related risk factors.

Donor-Specific Antibody Mean Fluorescence Intensity Threshold Predicts Platelet Transfusion Response in HLA-Alloimmunized Patients

Introduction:Patients with platelet transfusion refractoriness (PTR) due to HLA-alloimmunization respond to HLA-matched platelets. However, 4/4 HLA-matched platelets or platelets without donor-recipient antigenic mismatches are rarely available. Further, in severe HLA-alloimmunization, it is difficult to find donor products which do not contain antigens to which the recipient has developed antibodies ( i.e. antibody specificity prediction, ASP method). Hence, studies have evaluated platelet transfusion responsiveness in the presence of ≥1 donor specific antibodies (DSAs) to widen donor pools, a strategy termed permissive mismatching (PERMM). Using established semi-quantitative techniques for HLA-antibody detection, we sought to identify a cumulative DSA threshold which predicted platelet corrected count increment (CCI) failure despite PERMM. Methods:We retrospectively reviewed platelet transfusions among patients with HLA-alloimmunization and suspected PTR at the University of Washington/Fred Hutchinson Cancer Center from 2021-2023. HLA Class I antibodies were detected using a solid-phase single antigen bead assay (One Lambda) on the Luminex platform with individual antibody strengths measured by the mean fluorescence intensity (MFI). The calculated panel reactive antibodies (cPRA) were defined as the proportion of donors against whom recipients had antibodies. Transfusions were categorized as HLA matched, PERMM (DSA MFI&amp;gt;0), or random donor platelets (HLA-type unknown). CCI was calculated as described by Cohn ASH 2020. Results: Of 420 patients suspected to have PTR, 75 had cPRA &amp;gt; 50% and received ≥3 HLA-selected platelet transfusions. Of the 2726 transfusions these patients received, 1198 were excluded due to unavailability of post-transfusion platelet counts within 2 hours or due to multiple transfusions occurring between platelet counts. Ultimately, 1528 transfusions were analyzed (Panel A). In 1031 PERMM transfusions, the cumulative MFI was a stronger predictor of CCI at 2 hours than cPRA or the number of mismatched DSA (coefficients of determination or R2 of 7.3%, 2%, and 0.4%, respectively). We then examined the spectrum of cumulative MFIs to identify a threshold MFI predicting transfusion success/failure among the PERMM products. PERMM platelet transfusions in recipients with DSA MFIs of up to 5000 generated CCIs similar to HLA-matched platelets. However, CCI was significantly different between MFI &amp;lt;10,000 (median CCI 17,783) and MFI ≥10,000 (median CCI 3753) (Panel B). We then stratified data by splenomegaly, a significant non-immunologic cause of PTR (23.2% of transfusions, R2 15.8% for CCI prediction). Of the transfusions not affected by splenomegaly, 9.6% did not generate an adequate platelet response (defined as a CCI &amp;gt;5000) across all MFIs. This failure rate increased to 33.3%, 40.8%, and 52.8% among transfusions in recipients with MFIs &amp;gt;5000, &amp;gt;7500, and &amp;gt;10,000, respectively. After multiple testing correction, MFI of several individual DSAs (A1, A2, A24, A68, B8, B37, B39, B44, B51, B57, and B62) were also found to be significant predictors of CCI using linear regression ( n ranging 32-193, R2 ranging 6-33%). Conclusions: This is the largest study to date evaluating optimal MFI thresholds of DSAs for permissive HLA mismatching in the context of platelet transfusions. Cumulative DSA MFIs demonstrated a dose response effect with lower MFIs generating better CCIs. In our patient cohort overall, and when stratified by splenomegaly, we identified a threshold for cumulative DSA MFIs of 10,000, under which &amp;gt;50% of transfusions resulted in an adequate CCI. This MFI threshold (&amp;gt;10,000) has been implicated in HLA mismatched solid organ transplant rejections (Kallon, Transf. Med. 2020) as well as in a pooled analysis of smaller HLA-selected platelet transfusion studies conducted by our group (unpublished). If incorporated into a transfusion strategy, this MFI threshold may perform better than units chosen based on the number of mismatched DSAs alone or random donor platelets and widen the donor pool. Identifying the impact of specific HLA antibodies on CCIs needs further study. Studies are also underway to correlate cumulative DSA MFIs with complement activation (C1q levels) as a possible mechanistic explanation for the differences noted across the MFI threshold of 10,000 in our study.

Visual art on psychological anxiety in film and television animation scene design

BackgroundThe negative emotions of anxiety can cause a decrease in the body’s immune system, and endocrine disorders, and lead to cardiovascular diseases. Therefore, improving psychological anxiety is of great necessity in the current social context.Subjects and MethodsTo explore the impact of visual art on psychological anxiety in the design of film and television animation scenes, 286 random platelet donors were selected and divided into a control group and an observation group. The observation group watched the selected film and television animations during platelet donation, while the control group did not watch the images. During the donation process, the anxiety scores of the two groups were recorded using the State Anxiety Inventory (SAI), and the heart rate and pain values of the two groups were recorded using a desktop sphygmomanometer and a linear visual analog scale (VAS).ResultsThere was no significant difference in physiological and psychological indicators among the control group, with an anxiety score of (42.02 ± 0.40) kPa, a heart rate of (72.46 ± 8.13) beats/min, and a pain score of (2.35 ± 0.66) points. The anxiety score of the observation group was (35.49 ± 7.67) kPa, the heart rate was (70.67 ± 8.46) beats/min, and the pain score was (1.98 ± 2.64) points.ConclusionsThe difference between the two groups was statistically significant (P&lt;0.05). Therefore, visual art in film and television animation scene design has a positive impact on psychological anxiety.

A pilot study of cord blood-derived natural killer cells as maintenance therapy after autologous hematopoietic stem cell transplantation.

Natural killer (NK) cell based immunotherapy is an emerging strategy in hematologic malignancies because allogeneic NK cells can provide potent antitumor immunity without inducing graft-versus-host disease. Thus, we expanded cord blood-derived NK (CB-NK) cells ex vivo from random (MHC mismatched and KIR mismatched) donors, and investigate the feasibility and efficacy of repeated infusions CB-NK cells as maintenance therapy after autologous hematopoietic stem cell transplantation (ASCT). Thirty-one patients with acute myeloid leukemia and high-risk lymphoma received ASCT and the adoptive CB-NK cell multiple infusions for maintenance therapy. Patients received a median dose of 5.98 × 107/kg (range, 1.87-17.69 × 107/kg) CB-NK cells and 23 patients completed four infusions, 8 patients received three infusions. Only mild infusion reactions occurred in 15.5% of 116 infusions. Compared to a contemporaneous cohort of 90 patients who did not receive NK cell therapy, the adoptive transfer of CB-NK cells as maintenance treatment showed a tendency of difference in decreasing the relapse rate between CB-NK group and control group (9.7% vs 24.4%). The patients who receiving NK cell infusions had a better PFS and OS than controls (4 year PFS, 84.4 ± 8.3% vs 73.5 ± 5.4%; and 4 year OS, 100% vs 78.1 ± 5.4%) . These findings demonstrate safety and validity of maintenance therapy using CB-NK cells multiple infusions after ASCT, and it is worthy of further clinical trial verification.

Decreased rate of blood donors with high ABO antibody titers in Japan and the underlying factors: Comparisons between 2010 and 2021

Background and ObjectivesPreviously (2007), it was reported that ABO antibody titers in Japanese blood donors had decreased significantly compared to 20 years before. Here we evaluated whether further decrease of antibody titers had occurred in recent years, and the potential factors associated with changes in antibody titers. Materials and MethodsSerum/plasma from random blood donors in 2010 and 2021 (2010: 3369, 2021: 5796 donors) was classified into low, middle, and high ABO antibody titers according to the reactivity of diluted serum/plasma (2.5-fold and 20-fold) by an automated microplate system. The rates of low/high titer in the two periods were compared. Logistic regression and age-gender-BMI subgroup analyses were conducted to identify the factors that contributed to changes in antibody titers. ResultsCompared to 2010, the rate of donors with high ABO antibody titers was decreased in 2021 for both anti-A and anti-B (anti-A, 2010: 23.8%, 2021: 19.3%; anti-B, 2010: 23.8%, 2021: 16.4%). In logistic regression analysis, age was found to significantly affect both anti-A and anti-B antibody titers (anti-A, adjusted odds ratio 0.36, 95% CI 0.31–0.41; anti-B, 0.42, 0.37–0.47), and BMI (0.82, 0.73–0.92) and other time-related factors (0.79, 0.71–0.88) significantly affect anti-B antibody titers. Subgroup analysis revealed decreased rate of high anti-B titers in the higher age group in 2021. ConclusionThe rate of high ABO antibody titers, especially high anti-B titers, was significantly decreased in 2021, and our results suggested an association with aging and obesity of blood donors as well as other time-related factors.