Blood Transfusion

Partial s antigen expression in GP(B-A-B) hybrid glycophorins and the corresponding in vitro expression study

Ling Wei — 2025-05-14

Background - The s antigen expression is mainly determined by a single nucleotide polymorphism at c.143C (p.Thr48) on the exon 4 of GYPB gene. Several mutations on the GYPB gene have been reported to cause aberrant s antigen expression. GP.Mur has an extra 31-amino acid insertion encoded by the active compound GYP(B-A) exon 3, which closely locates at the upstream of p.Thr48. It has been reported to cause altered s antigen expression.

Materials and methods - Serologic testing and flow cytometry analysis were performed to detect s antigen expression on RBCs of GP(B-A-B) hybrid glycophorins, including GP.Mur, GP.Bun and GP.HF. Several mutant plasmids based on the different sites between GYPB and GYP(B-A-B) alleles were constructed and transfected into HEK293T cells for in vitro expression, to reveal the key amino acids for the aberrant s antigen expression.

Results - Serologic testing and flow cytometry assay showed the RBCs of GP.Mur homozygotes reacted positively with IgG anti-s (P3YAN3) but negatively with IgM anti-s (P3BER). Flow cytometry analysis also revealed half level of s antigen expressed on the RBCs of GP.Mur, GP.Bun and GP.HF heterozygotes with ss genotype compared to S-s+ controls when detected by IgM anti-s (P3BER). Furthermore, in vitro expression study showed that p.Asn45 is critical for the epitope expression of s antigen detected by IgM anti-s (P3BER).

Discussion - The results demonstrated partial s antigen expression on GP(B-A-B) RBCs. In addition to p.Thr48, p.Asn45 is also important for the epitope expression of s antigen detected by IgM anti-s (P3BER). To avoid false negative serologic typing, it is recommended to use several different clones of monoclonal anti-s for the correct s typing, especially in the regions with high frequency distribution of GP(B-A-B) hybrid glycophorins.

The impact of red cell storage age on transfused patients with sickle cell disease: protocol of a pilot randomized clinical trial to evaluate changes in inflammation and clinical transfusion efficacy

Matthew Karafin — 2025-02-04

Background - Despite fulfilling all requirements for donor blood units as defined by the FDA, a number of patients with sickle cell disease (SCD) are transfused with red blood cell (RBC) units that are near the end of their storage life, exposing them to the potentially adverse components of the red cell storage lesion. Due to their chronically inflamed state, patients with SCD may be particularly susceptible to these components. We present here a pilot study protocol for testing the impact of fresh vs older red cell units in chronically transfused adults with SCD.

Materials and methods - This is a randomized, prospective, clinical trial. We aimed to recruit forty chronically transfused adults or adolescents with SCD who receive regular RBC transfusions for their clinical care and randomize these patients to receive either units greater than or equal to 30 days, or units less than or equal to 10 days for 3 consecutive outpatient transfusion events.

Results - The primary endpoint is the metabolic differences identified between units transfused that are greater than or equal to 30 days, and those units less than or equal to 10 days. The secondary endpoint evaluates the change in blood monocyte activation at 2 hours after transfusion between the two groups. Lastly, weevaluate unit RBC efficacy via changes in
hemoglobin/day, hemoglobin A%/day, hospitalization rate, pain scores, and infections as documented via blood and urine cultures.

Discussion - This study promises to provide evidence as to whether metabolically older red cell units affect the quality and efficacy of chronic transfusion therapy for adults with SCD and has the potential to guide the need for future study on this important clinical issue.

Patient Blood Management in pediatric and adolescent bone marrow donors: results from an Italian survey

Claudia Del Fante — 2024-11-28

Background - Current national and international guidelines (Italian Bone Marrow Donor Registry [IBMDR], World Marrow Donor Association [WMDA] standards) provide an indication for preoperative autologous blood donation (PAD) only in adult family and volunteer non-family donors in anticipation of bone marrow (BM) hematopoietic stem cell (HSC) donation to avoid the use of homologous transfusions. In addition, there is no clear guidance from the relevant scientific societies regarding pediatric and adolescent donors.

Material and methods - To assess the actual use of PAD in pediatric (up to 14 years) and adolescent (aged 15-18 years) family donors in relation to BM HSC donation in the five years 2017-2021, a specific online questionnaire was administered to blood establishments and clinical units of pediatric transplantation programs responsible for BM HSC collection.

Results - Adherence to the project was 100% (18/18 centers). During the five-year period considered, 273 BM HSC donors (205 pediatric and 68 adolescent) were registered. Forty percent of the non-trait carrier donors who underwent PAD received iron therapy in preparation for BM HSC donation; only 4.8% of the pediatric and none of the adolescents had hemoglobin values below the age limit at donation. Finally, 66.4% of pediatric donors and 15.4% of non-trait carrier adolescent donors who did not undergo PAD received homologous transfusions during BM harvest.

Discussion - The present study highlights the highly heterogeneous criteria for the use of PAD (including calculating of the volume of whole blood collected) and the lack of a specific policy in preparation for BM HSC donation, either from non-trait carrier donors or those with sickle cell or thalassemia trait, both pediatric and adolescent.

Impact of leukoreduction on the metabolome of ovine packed red blood cells during refrigerated storage

Arianna Miglio — 2025-02-06

Background - Blood transfusion is a life-saving intervention for many species of veterinary interest, including sheep. Despite extensive research on the impact of refrigerated storage of packed red blood cells (pRBC) in humans, research on the quality of stored ovine blood is limited and storage guidelines are mostly informed by studies in humans. Human pRBC are currently stored without residual white blood cells, following selective removal of the leukocytes by filtration (leukoreduction). This process delays the onset and mitigates the progression of the storage lesion, a series of molecular changes that RBC undergo as a function of storage duration. However, leukoreduction of ovine pRBC is not routinely performed.

Materials and methods - Here we performed metabolomics analyses of
non-leukoreduced (nLR) and LR pRBC from six sheep. Units were stored under standard veterinary blood bank conditions (4°C) for up to 42 days and sterilely sampled weekly for metabolomics analyses of cells and supernatants.

Results - LR-pRBC showed significantly lower levels of mono-, di- and
tri-carboxylates in both the cellular and supernatant compartments, and slower accumulation of lactate and immunomodulatory succinate, fumarate and malate. The presence of residual white blood cells in the units accelerated the consumption of glucose from the media, with no increase in detectable high energy phosphate compounds (AMP). nLR showed a higher degree of purine breakdown and deamination products, (hypoxanthine, xanthine and allantoate). Elevated free fatty acids in nLR RBC are consistent with increased lipid peroxidation and lipolysis. Strong sex dimorphism was observed across all samples, independently of storage duration or leukoreduction.

Discussion - Leukoreduction of ovine pRBC delays the onset and mitigates the metabolic storage lesion to central energy and redox metabolism, while almost completely abrogating the accumulation of carboxylates in stored units.

The tandem CD33-CLL1 CAR-T as an approach to treat acute myeloid leukemia

Huiru Wang — 2024-08-06

Background - Acute myeloid leukemia (AML) is characterized by high heterogeneity, poor long-term survival, and a propensity for relapse. Exceptional efficacy in treating recurrent or refractory B-lymphoid malignancies has been demonstrated by Chimeric antigen receptor T cells (CAR-T cells). Given the therapeutic potential of targeting both CD33 and C-type lectin-like molecule-1 (CLL1) in AML, the development of a dual-targeting CD33-CLL1 CAR-T cells assumes significant importance.

Materials and methods - The expressions of CD33 and CLL-1 antigens in peripheral blood cells and bone marrow cells from AML patients was assessed. Subsequently, a Chimeric Antigen Receptor (CAR) incorporating a dual-specific single-chain variable fragment targeting CLL1 and CD33 (CD33-CLL1-CAR-T) was engineered. The anti-tumor efficacy and potential side effects of CD33-CLL1-CAR-T cells were comprehensively investigated in both in vitro and in vivo settings.

Results - The constructed tandem CD33-CLL1 CAR-T exhibited potent cytotoxicity against leukemia cell lines and human primary AML cells in vitro. Co-cultivation of AML blasts with CD33-CLL1-CAR-T cells resulted in effective proliferation and the secretion of substantial quantities of GM-CSF and IFN-γ. Importantly, the impact of CD33-CLL1-CAR-T cells on normal hematopoietic stem cells was minimal, ensuring safety in vivo mouse models. Notably, significant anti-leukemic activity was observed in the mouse model, with CD33-CLL1-CAR-T cells leading to tumor eradication and prolonged survival.

Discussion - The tandem CD33-CLL1 CAR-T cells not only efficiently eliminated AML blasts but also exhibited low cytotoxicity toward normal hematopoietic stem cells (HSCs). These findings underscore the potential clinical applicability of the tandem CD33-CLL1 CAR-T cells as an effective and safe treatment strategy for AML, representing a noteworthy advancement in the field of CAR-T cells therapy.

First use of cord blood platelet-rich plasma in the treatment of vulvar lichen sclerosus: a preliminary study towards a randomized controlled trial

Veronica Boero — 2024-12-17

Background - Although topical corticosteroids (TCS) represent first-line treatment for vulvar lichen sclerosus (VLS) and as such should be prescribed to all women at time of diagnosis, approximately 30% of patients do not experience complete symptom resolution following such treatment. TCS may not effectively improve vulvar trophism and elasticity, both of which are crucial for sexual function. Owing to its regenerative and healing properties, cord blood platelet-rich plasma (CB-PRP) may represent an efficacious supplementary therapy, to be administered following first line treatment with TCS. The primary aim of this study was to assess safety and tolerability of
CB-PRP in women with VLS.

Materials and methods - This is a pilot study which precedes a randomized controlled trial of CB-PRP vs placebo in women with VLS. Ten consecutive patients with VLS, who had previously undergone standard TCS-treatment, received three vulvar CB-PRP injections monthly. Follow-up was conducted three months after the last injection using vulvoscopy and validated questionnaires to evaluate safety and tolerability, as well as patient satisfaction, symptom improvement, sexual function, psychological well-being, quality of life, frequency of TCS application as a maintenance treatment, vulvar trophism and architectural modifications.

Results - No adverse clinical effects were observed. Five patients (50%) were either satisfied or very satisfied with the procedure, four (40%) were uncertain about their satisfaction with the treatment. One patient (10%) dropped out for personal reasons and was classified as unsatisfied according to an intention-to-treat analysis. At follow-up median numeric rating scale scores were significantly reduced for vulvar burning compared to baseline (p<0.05) there was a trend toward improvement in itching, dyspareunia, and dysuria. A significant improvement in sexual arousal and satisfaction was observed in all treated women (p<0.05).

Discussion - CB-PRP may be a promising treatment for VLS. It appears to be safe and improve symptoms and sexual function.

Platelet rich plasma for assisted reproduction: an overview of systematic reviews

Francesca Masiello — 2025-04-15

Background - In recent years, platelet-rich plasma (PRP) has been used to improve endometrial receptivity in infertile women undergoing assisted reproduction. This article aims to review the literature and critically appraise the available evidence regarding the efficacy of PRP for assisted reproduction.

Materials and methods - An overview of systematic reviews (SRs) (umbrella review) of PRP use for assisted reproduction. The methodological quality of the SRs was assessed using the AMSTAR-2 checklist; quality of the evidence from the trials included in each SR was appraised following the GRADE approach.

Results - Twenty-five SRs published between 2020 and 2024 were included in this overview. The SRs reported data from 306 overlapping reports based on 112 individual primary studies. The overlap among primary studies was slight across all reports included (CCA index 1.9%). With the AMSTAR 2 tool, the SRs showed only some non-critical weakness, but no critical flaws. Thus, confidence in the results of the SRs can be considered high or moderate. Clinical pregnancy rate was reported in 17 SRs, chemical pregnancy in 8, implantation rate in 8, and live birth rate in 16 SRs. Almost uniformly the SRs showed increased rates of positive outcomes in PRP recipients compared to controls (from very low to high certainty of evidence). Miscarriage rate, ectopic pregnancy and multiple pregnancy, reported in some of the SRs, resulted uncertainty about the effect of PRP infusion compared with no intervention on these outcomes (no statistically significant differences between groups).

Discussion - The evidence from SRs shows beneficial effects of PRP compared to controls in terms of clinical and chemical pregnancy rates, implantation rates and live birth rates. The level of evidence varied from very low to high (more commonly low or moderate) in relation to methodological flaws and clinical heterogeneity of primary studies included in the SRs.

Combination of Evans syndrome and COVID-19: a systematic review of reported cases

Suwen Li — 2025-02-06

Background - Evans syndrome is a rare autoimmune disease characterized by simultaneous or sequential primary immune thrombocytopenia and autoimmune hemolytic anemia. Despite the low incidence of Evans syndrome after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, its progression may threaten public health. This review offers an up-to-date summary of the works on the association between coronavirus disease 2019 (COVID-19) and Evans syndrome to explore the pathogenic mechanisms, epidemiological characteristics, clinical presentations, diagnostic markers, and treatment strategies.

Material and methods - We searched PubMed and Web of Science to identify articles that explored the relationship between COVID-19 and Evans syndrome. We collected and organized all reported cases of Evans syndrome following COVID-19 or SARS-CoV-2 vaccination over the past 4 years and also expanded the search to examine other cases of post-infection Evans syndrome.

Results - Thirteen cases were included with an average age of 42 years of whom 12 survived and one died. Two cases were associated with pregnancy and four with vaccination, two involved epileptic seizures, and three had a history of autoimmune disease.

Discussion - Patients with Evans syndrome and exposure to SARS-CoV-2 have a potential risk of bleeding. This risk should prompt close monitoring of bleeding biomarker dynamics and early initiation of hemostatic treatments, including platelet transfusion, corticosteroids, thrombopoietin receptor agonists, intravenous immunoglobulin and rituximab.

Hemostasis testing practices vary in patients with high bleeding risk undergoing therapeutic plasma exchange

Alexandre Soares Ferreira Junior — 2025-05-09

In high bleeding risk patients undergoing therapeutic plasma exchange (TPE), the optimal hemostasis testing strategy is not known. Therefore, in a cohort of patients with a major bleeding event prior to TPE, we sought to describe hemostasis testing practices. In a cross-sectional analysis of the Recipient Epidemiology and Donor Evaluation Study-III public use data files, we identified patients with bleeding prior to TPE initiation. Within two days prior to TPE initiation, we assessed performance of any of the following tests: Platelet count, International Normalized Ratio (INR), activated Partial Thromboplastin Time (aPTT), and Fibrinogen. Results were stratified by bleeding recurrence status (Yes vs No recurrence). Bleeding recurrence after TPE was defined as a major bleeding event occurring within two days of a TPE procedure. Among 310 patients meeting inclusion criteria, hemostasis tests were done in 298 (96%). For all patients, the most common hemostasis test was the platelet count (95%). The least common was fibrinogen (36%). Among the Yes recurrence (No.=121) and No recurrence (n=189) cohorts, the most common test combinations were Platelet + INR (68% and 77%, respectively) and Platelet + INR + aPTT (78% and 71%, respectively). Across both cohorts, all four hemostasis tests were assessed in 33% and 22%, respectively. In patients with bleeding prior to TPE, hemostasis tests are assessed inconsistently. The effectiveness of hemostasis testing in both identifying and mitigating bleeding risk should be assessed in future studies.

Identification of a novel ABO*O.01.01 allele with c.801G>T mutation in a Chinese A2 subtype individual

Chonghe Xu — 2024-11-29

The presence of ABO subgroup alleles and abnormal O alleles is often associated with inconsistent serologic findings in ABO blood group typing. ABO phenotypes were identified by standard serological agglutination tests and allele direct sequencing and haplotype sequencing analyses of the ABO gene were carried out. The serologic results showed the female case was preliminarily assigned as A2 subtype. The sequencing results showed the patient was heterozygous for a novel O allele (c.801G>T) and A205, which caused a p.Gly267Gly synonymous variant.