Donor Samples Research Articles

Identification of near full-length human pegivirus type 2 (HPgV-2) genomes in blood donor samples co-infected with hepatitis C virus (HCV).

Human pegivirus (HPgV) identified from an HCV-infected plasma sample through nanopore metagenomics. The analysis revealed a nearly complete HPgV-2 genome. Phylogenetic analysis confirmed its classification within the HPgV-2 genotype, providing insights into viral co-infection dynamics. The metagenomics approach advances the understanding of viral diversity and supports the development of accurate diagnostics.

Prospective analysis of biomarkers associated with successful faecal microbiota transplantation in recurrent Clostridioides difficile Infection.

Faecal microbiota transplantation (FMT) is an established treatment for recurrent Clostridioides difficile infection (R-CDI). This study aimed to identify calprotectin and microbiome characteristics as potential biomarkers of FMT success. We conducted a prospective study of patients who underwent oral FMT (single dose of 4-5 capsules) for R-CDI (January 2018 to December 2022). Samples were collected at three time points: at CDI diagnosis, within 24hours prior to FMT administration, and 30days post-FMT. Calprotectin levels were assessed and the V4 region of the 16S rRNA gene was sequenced to analyse the microbiota composition. Sequencing data analysis and statistical analysis were performed using MOTHUR and R. Ninety-seven patients underwent FMT (totalling 105 procedures). A total of 221 samples were processed, including 21 donor samples, 24 capsule contents, and 176 patient faecal samples (39 at diagnosis, 63 pre-FMT, and 74 post-FMT). FMT achieved an overall success rate of 85.1% (86/101 cases). The abundance of Bacteroides, Ruminococcus, Megamonas, and certain Prevotella operational taxonomic units (OTUs) was significantly higher in capsules associated with 100% success compared to less effective capsules. FMT engraftment was observed in 95% of patients with favourable outcomes versus 62% of those with recurrences (p = 0.006). Additionally, a negative correlation was found between calprotectin levels and specific microbial genera, suggesting an association with successful outcomes. This study highlights differences in the evolution of faecal microbiota, bacterial engraftment, and inflammation markers (e.g., calprotectin) between patients with varying FMT outcomes. Potential biomarkers for successful FMT were identified, providing valuable insights for optimizing FMT strategies.

Evaluation of immunoglobulin M and immunoglobulin G titers of ABO blood group system in blood donors at a blood center of a tertiary care hospital

Abstract BACKGROUND AND OBJECTIVES: ABO antibodies have implications in platelet transfusions, solid organ, and bone marrow transplants. The study aimed at measuring the immunoglobulin G (IgG) and immunoglobulin M (IgM) ABO antibody titers in A, B, and O blood group blood donors and correlating the titers with age, gender, and blood group of donors. SETTINGS AND DESIGN: A prospective observational study was performed over 3 years at a tertiary care blood center. MATERIALS AND METHODS: Equal number of A, B, and O group blood donor samples chosen by random sampling were evaluated for titers by doubling dilution using tube method. IgM titers were determined at room temperature. IgG titers were tested at 37°C with anti-IgG antiglobulin reagent after inactivating IgM antibodies by dithiothreitol. The titers were correlated with age, gender, and blood group. STATISTICAL ANALYSIS: Mann–Whitney U-test, Kruskal–Wallis test, and Bonferroni correction were employed for various correlations using IBM SPSS v26 (Statistical Package for the Social Sciences) software. A two-tailed P < 0.05 was considered statistically significant. RESULTS: A total of 870 samples (290 of each of A, B, and O blood groups) were tested for IgG and IgM antibody titers. IgG anti-A and anti-B titers were significantly higher in blood group O as compared to blood groups B and A. Anti-B IgM titers in group A were significantly higher than anti-A IgM titer in B. The antibody titer showed decreasing trend with age. There was no correlation with gender. CONCLUSIONS: The study will help hospitals and blood centers in formulating policies regarding ABO antibody titer estimation in platelet donors and renal transplant recipients in the context of out-of ABO group platelet transfusions and ABO-incompatible renal transplants, respectively.

DOP020 Baseline Microbiota and Differences in Donor-Recipient Richness as Determinants of Fecal Microbiota Transplantation Efficacy in Ulcerative Colitis

Abstract Background Rigorous donor preselection, strict anaerobic processing, and repeated faecal microbiota transplantation (FMT) administration did not improve outcomes for induction of clinical remission in the RESTORE-UC trial1. We studied the luminal, and mucosal microbiota composition as well as gene expression throughout the RESTORE-UC study, to further understand the results and instruct future FMT trial design. Methods The RESTORE-UC trial was a multi-centric double-blind, sham-controlled randomized trial. Patients with moderate to severe UC (total Mayo 4-10) were randomly allocated to receive 4 anaerobic-prepared allogenic or autologous donor FMT. Mucosal biopsies were collected for RNA- and 16S sequencing at week 0 and 8. Quantitative microbiota profiling was performed on donor samples, and in patients at weeks 0, 1, 2, 3, 4, 8, 12, 26, and 52 (n=756). Clinical success (steroid free clinical remission) and -response (≥3 points or ≥50% reduction in combined Mayo subscores for rectal bleeding and stool frequency) were evaluated at week 8. Metagenomic success (week 8) was defined as restoration of eubiosis (Bacteroides 2 (B2) to non-B2 enterotype) and failure was considered with subtypes being no transition, non-B2 to other non-B2 transition, lead dysbiosis (non-B2 to B2), and maintenance of dysbiosis (B2 to B2). Results Baseline beta diversity (metric for overall microbial similarity) was significantly associated with future clinical and metagenomic success (resp. p=0.035; p=0.003). Similar trends were observed at week 8 (resp. p=0.056; p=0.001). Restoration of eubiosis was characterized by a rapid increase in bacterial richness after the first FMT administration, which was maintained up to 12 months after intervention (Fig A). A greater difference (Fig B) in donor-recipient richness determined the metagenomic success (p=0.06) and clinical response (p=0.01), while this was not observed for clinical success (p=0.16). Metagenomic success was accompanied by increased engraftment after FMT (Fig C, p<0.01). Independently from clinical success, patients with dysbiotic faecal communities at baseline had distinctive mucosal communities in their intestinal biopsies (p<0.01), which were significantly changed in samples where eubiosis was restored (p=0.012), but not in those where clinical success was observed (p=0.34). However, this could be explained by the low overall response rate. No specific transcriptomic profiles in mucosal biopsies at baseline were reminiscent of metagenomic (p=0.24), nor clinical success (p=0.22). Conclusion Patient and donors should be rigorously selected in future FMT trials, and selection should be based on microbial composition and by obtaining a great difference in richness between receptor and donor. References 1 Caenepeel*, Deleu*, Vazquez* et al., 2024

P0270 Standardisation of Rapid TDM Assay with WHO International Reference Material for Anti-Infliximab Antibodies

Abstract Background Point-of-care (POC) tests play a crucial role in therapeutic drug monitoring (TDM) for Inflammatory bowel disease (IBD) patients undergoing anti-TNF therapy. Infliximab (IFX), the first biologic used in IBD treatment, is a chimeric anti-TNF-alpha blocker. However, its composition can trigger an immune response in patients, leading to the production of anti-drug antibodies (ADAs). Consequently, regular monitoring for ADAs is essential during IFX therapy. Various assays are available to measure anti-IFX antibody levels, mainly based on ELISA technology. The recent introduction of an international reference material (IRM) for infliximab ADAs allows enhanced measurement traceability and enables harmonised standardisation and improved comparability across different assays. Furthermore, it is mandatory to standardise with an existing IRM for compliance with the in vitro diagnostic regulation (IVDR). This study focused on standardising the lateral flow based Quantum Blue® Anti-Infliximab assay (LF-ADIF) using the IRM and comparing its performance with the current standardisation material. Additionally, a method comparison to established ELISA assays was carried out. Methods The lateral flow assay was calibrated with both the currently used standardisation material and the new IRM. Blant-Altman analysis was performed to evaluate the comparability of both standardisation materials. To establish the cut-off, 105 healthy donor samples were measured. This cut-off will be used for discrimination of positive and negative results of the rapid test. Furthermore, a set of 30 patient samples was analysed for method comparison with established ELISA assays. Results The Blant-Altman analysis revealed that a IRM based calibration is feasible and IRM concentrations <100 IU/mL can be detected. A technical cut-off was established allowing the discrimination of positive and negative patient samples. Total agreement between the lateral flow assay and the ELISA assays was demonstrated in the method comparison. Conclusion To the best of our knowledge the here presented assay is the first IRM based and IVDR ready lateral flow assay for anti-infliximab. The total agreement of patient sample results between the Quantum Blue® Anti-Infliximab assay and commercially available ELISA assays revealed good comparability. IRM standardisation across all anti-IFX assays would further improve comparability between them.

Optimizing mouse metatranscriptome profiling by selective removal of redundant nucleic acid sequences.

Sequencing total RNA from microbiome samples is seriously impaired by the overwhelming proportion of rRNA to mRNA content. As much as 99% of sequencing reads can be assigned to the rRNA content, thus removal of these abundant transcripts is critical to MetaT analysis. The use of Ribo Zero Plus rRNA depletion probes designed for human gut microbiomes proved to be less effective and more inconsistent across mouse cecal donor samples, a common experimental system for microbiome studies. In the present work, we have extended and refined a taxonomically-neutral probe design method for mouse cecal content. The additional probes were carefully chosen to limit the number needed for effective depletion to reduce both the cost and risk of introducing bias to MetaT analysis. Our results demonstrate this method as efficient and consistent for rRNA removal in mouse cecal samples, thus providing a significant increase in the number of mRNA-rich sequencing reads for MetaT analysis.

The effects of age and other individual factors on radiation induced ESR signals from fingernails

Biodosimetry is crucial for assessing ionizing radiation exposure to guide medical responses. Electron spin resonance (ESR) spectroscopy using fingernails can be effectively used for both occupational and public dose assessments in radiological accidents because of their accessibility and ability to retain stable radiation-induced free radicals. However, despite two decades of research, challenges remain in achieving accurate fingernail dosimetry, mainly owing to the variation in ESR signals among individuals. The purpose of this study was to explore inter-individual differences in ESR signals in fingernails to improve the accuracy and reliability of extremity dosimetry. Fingernail samples were collected from 15 participants (age: 11–64 years), irradiated with X-rays (160 kV, 6.3 mA) at 0, 5, 10, and 20 Gy, and measured using ESR spectroscopy. The effects of individual factors, such as age, sex, health condition, and lifestyle, on radiation-induced ESR signals (RIS) were investigated. Younger participants exhibited stronger RIS intensities and a more linear dose–response relationship. The RIS intensity in female samples tended to be higher than that in male samples. Interestingly, the fingernals of middle-aged donors who regularly took vitamin supplements showed significantly higher ESR signal intensities than those of similar-age donors who did not take supplements. Notable reductions in RIS intensity during storage in a freezer were observed only in older donor samples irradiated at higher doses. These findings underscores the importance of considering age and other individual factors in the calibration for fingernail dosimetry.

Harnessing gut microbial communities to unravel microbiome functions.

The gut microbiome impacts human health in direct and indirect ways. While many associations have been discovered between specific microbiome compositions and diseases, establishing causality, understanding the underlying mechanisms, and developing successful microbiome-based therapies require novel experimental approaches. In this opinion, we discuss how in vitro cultivation of diverse communities enables systematic investigation of the individual and collective functions of gut microbes. Up to now, the field has relied mostly on simple, bottom-up assembled synthetic communities or more complex, undefined stool-derived communities. Although powerful for dissecting interactions and mapping causal effects, these communities suffer either from ignoring the complexity, diversity, coevolution, and dynamics of natural communities or from lack of control of community composition. These limitations can be overcome in the future by establishing personalized culture collections from stool samples of different donors and assembling personalized communities to investigate native interactions and ecological relationships in a controlled manner.

Potential Regulation of ARID1A by miR-129-5p and miR-3613-3p and Their Prognostic Value in Gastric Cancer.

Gastric cancer (GC) remains the most common malignant tumor of the gastrointestinal tract and one of the leading causes of cancer-related deaths worldwide. Non-coding RNAs (ncRNAs), including microRNAs (miRNAs), are involved in the pathogenesis and progression of GC and, therefore, may be potential diagnostic and prognostic biomarkers. Our work was aimed at investigating the predicted regulation of ARID1A by miR-129-5p and miR-3613-3p and the clinical value of their aberrant expression in GC. The study included tumor and adjacent non-tumor tissues from 110 GC patients, 38 sectional normal gastric tissue samples, as well as 65 plasma samples of GC patients and 49 plasma samples of healthy donors. Expression levels of ARID1A and both miRNAs were quantified by reverse transcription-polymerase chain reaction (RT-PCR). We have identified significant associations of their expression with the clinical and pathological characteristics of GC patients both in tissues and plasma. To validate predicted target pairs miR-129-5p/ARID1A and miR-3613-3p/ARID1A, in vitro experiments on cancer cell lines were conducted. The obtained results suggest a complex role of ARID1A, miR-129-5p and miR-3613-3p in GC and potential regulation of ARID1A expression by both miRNAs.

Quantifying the effect of human interindividual kinetic differences on the relative potency value for riddelliine N-oxide at low dose levels by a new approach methodology.

Pyrrolizidine alkaloids N-oxides (PA-N-oxides) are predominant in plants and herbal foods, and are converted to pyrrolizidine alkaloids (PAs) upon consumption, leading to toxicity. The effect of interindividual kinetic differences on the relative potency values of PA-N-oxides compared to their PAs (REPPANO to PA) was studied, with riddelliine N-oxide (RIDO) and riddelliine (RID) as model compounds. In vitro kinetic data measured in incubations with 30 fecal and 25 liver S9 donor samples showed high variation across individuals, where the interindividual variability was captured with Bayesian multilevel regression. The distributions of influential PBK model parameters were used as input for physiologically based kinetic (PBK) modeling combined with Monte Carlo (MC) simulations to calculate the probability distribution of REPRIDO to RID values. At low internal dose levels, interindividual differences were shown to be a factor that influences the REPRIDO to RID value while neither dose nor endpoint used plays a role. The distribution of the REPRIDO to RID value ranged from 0.71 to 0.97 (95th percentile) with a mean value of 0.87. The approach described enables determination of interindividual REPPANO to PA values at low dose levels, which are not accessible in in vivo experiments quantifying the REP value.

SnRNA-seq stratifies multiple sclerosis patients into distinct white matter glial responses.

Poor understanding of the cellular and molecular basis of clinical and genetic heterogeneity in progressive multiple sclerosis (MS) has hindered the search for new effective therapies. To address this gap, we analyzed 632,000 single-nucleus RNA sequencing profiles from 156 brain tissue samples of MS and control donors to examine inter- and intra-donor heterogeneity. We found distinct cell type-specific gene expression changes between MS gray and white matter, highlighting clear pathology differences. MS lesion subtypes had different cellular compositions but surprisingly similar cell-type gene expression patterns both within and across patients, suggesting global changes. Most gene expression variability was instead explained by patient effects, allowing us to stratify patients and describe the different pathological processes occurring between patient subgroups. Future mapping of these brain molecular profiles with blood and/or CSF profiles from living MS patients will allow precision medicine approaches anchored in patient-specific pathological processes.

The Umbra and the Penumbra: Longitudinal Effects of Geographic Atrophy in AMD on the Outer Choroid by Imaging Analysis and Histopathological Correlation.

To quantitate regional changes in the outer choroidal vessels in patients with geographic atrophy (GA) in AMD and to correlate with a histopathological donor sample. We analyzed 35 participants with GA for in vivo analysis and 1 participant with subject for histopathological analysis. Participants underwent three structural optical coherence tomography scans spaced 6 months apart over 1 year. Quantitative measurements of the outer choroidal vessels were performed in three regions: the GA region, a 150-µm-wide border surrounding the GA, and the area beyond the border. Histopathological analysis was performed using europaeus agglutinin lectin-stained choroidal flat-mount images with the focal planes of both the choriocapillaris and the outer choroidal vessels. In the GA region, the median vessel area was 3390 µm2 (interquartile range [IQR], 2821 µm2) at baseline, 3139 µm2 (IQR, 2888 µm2) at the 6-month visit and 2888 µm2 (IQR, 2617 µm2) at the 12 month visit (P < 0.001). Our cohort was divided into two subgroups based on RPE atrophy development at the GA border at the 6-month visit. This analysis showed that significant choroidal shrinking occurred only in eyes where the GA border progressed to GA at the 6-month follow-up. Histopathological analysis also demonstrated loss of the outer choroidal vessels, which were predominant in the GA region. The outer choroidal vessels seem to decrease with time within the area of GA. The outer vessels decrease over time in eyes where the GA progresses. This finding may suggest that these vessels are under a very tight paracrine control.

2.3 – 00078 Detection of HIV-1 antisense transcripts in donor samples before and during ART

2.3 – 00078 Detection of HIV-1 antisense transcripts in donor samples before and during ART

Serological investigations on penicillin‐induced antibodies in the Thai population

AbstractObjectivesThe study aimed to assess the conditions for coating RBCs with penicillin and examine the anti‐penicillin reactions of random Thai patients' sera against penicillin‐coated RBCs and normal sera from Thai donors testing in the presence of the drug.BackgroundPenicillin‐induced immunologic haemolytic anaemia (IHA) is reportedly related to possessing antipenicillin antibodies, immunoglobulin G (IgG), which has been identified in testing penicillin‐coated red blood cells (RBCs). In addition, low titre penicillin antibodies, often IgM, are detected in donors by testing in the presence of a solution of the penicillin.Materials and MethodsPenicillin‐coated RBCs were produced, and antipenicillin was tested against those penicillin‐coated RBCs amongst random Thai patients who had strongly positive direct antiglobulin (≥3+). Additionally, sera from Thai blood donors were tested in the presence of the penicillin. These relationships were determined by comparing the numbers of penicillin‐antibody positive patients with their diagnosis, sex, age and blood type.ResultsPenicillin requires a high pH to optimally adhere to RBCs that showed validated reactions with controls. Enrolment of 304 random patients, of whom 17 (5.59%) had positive antipenicillin tests using penicillin‐coated RBCs. Of the 246 donor samples, 3 (1.22%) displayed positive reactivities in the presence of soluble penicillin. Furthermore, no association was discovered between the patient's characteristics and antipenicillin positivity.ConclusionsThis is the first study to develop and report on the low percentage of patients' and donors' sera without IHA. Investigating suspected cases of penicillin‐induced IHA requires following our suggested method to identify clinically significant antipenicillin.

Urgent Alert: Potential Risk of Dengue Infection Transmission Through Blood Transfusion in Iran

Dengue infection is an emerging public health issue in Iran, with about 149 confirmed newly infected cases. It can be transmitted by the bite of infected Aedes mosquitoes and even nosocomial routes. Due to the rapid replication and geographical spread of the mosquito, there is a potential risk of increased infected individuals. Given the possibility of the transmission of dengue infection through transfusion, it is important to implement policies to improve blood safety. Proper donor selection by utilizing appropriate blood donor questionnaires and performing general physical examinations, along with performing sensitive diagnostic tests on blood donor samples, utilizing pathogen reduction techniques, and implementing lookback programs, can be effective in reducing the risk of transfusion-transmitted dengue virus (TT-DENV).

Isolation and analysis of residual leucocytes from leucoreduced red blood cell units.

Leucoreduction is used to remove donor leucocytes during red blood cell (RBC) manufacture. However, not all are removed, and long-term survival of donor leucocytes, termed transfusion-associated microchimerism (TAM), has been shown to occur in some patients following RBC transfusion. The mechanism of TAM occurrence is unknown. One hypothesis is that viable donor haematopoietic cells remain within RBC units that could engraft. However, the analysis of cells remaining within leucoreduced RBC units has been minimal. This study aimed to isolate and analyse any residual leucocytes recovered from leucoreduced RBC units. Leucoreduced RBC units were analysed on Day 1 (n = 4) and Day 42 (n = 4) post collection. Residual leucocytes were isolated using the EasySep™ RBC Depletion Reagent. Cell type analysis was conducted by flow cytometry using a leucocount reagent, a viability marker (7-amino-actinomycin D [7AAD]) and specific antibodies to CD45 and CD34. A representative 'pre-filter' sample was also obtained at the time of whole-blood donation to ensure expected cell counts across the donor samples. Analysis of the pre-filter sample showed that CD45+/CD34+ cells accounted for 0.02%-0.07% of all leucocytes. Up to 253,850 residual leucocytes were isolated across both storage timepoints, and of these, up to 48 cells were CD45+/CD34+/7AAD-. Viable CD45+/CD34+ cells were isolated from leucoreduced RBC units, indicating the potential for donor progenitor cells to be present during transfusion. Further characterization of these residual cells is required to explain how TAM may occur in some patients following RBC transfusion.

Barcoding of viable peripheral blood mononuclear cells with selenium and tellurium isotopes for mass cytometry experiments.

Barcoding viable cells combined with pooled sample staining is an effective technique that eliminates batch effects from serial cell staining and facilitates uninterrupted data acquisition. We describe three novel and isotopically pure selenium-containing compounds (SeMals) that are useful cellular labeling tools. The maleimide-functionalized selenophenes (76SeMal, 77SeMal, and 78SeMal) covalently react with cellular sulfhydryl groups and uniquely label cell samples. The SeMal reagents label viable and paraformaldehyde-fixed peripheral blood mononuclear cells (PBMC), are well resolved by the mass cytometer, and have little spill into adjacent channels. They appear non-toxic to viable cells at working concentrations. We used SeMal reagents in combination with four isotopically pure tellurium maleimide reagents (124TeMal, 126TeMal, 128TeMal, and 130TeMal) to label 21 individual PBMC samples with unique combinations of selenium and tellurium isotopes (seven donors with three replicates using a 7 isotope pick 2 combinatorial schema). The individually barcoded samples were pooled, stained with an antibody cocktail as a pool, and acquired on the mass cytometer as a single suspension. The single-cell data were de-barcoded into separate sample-specific files after data acquisition, enabling an uninterrupted instrument run. Each donor sample retained its unique phenotypic profile with excellent replicate reproducibility. Unlike current live cell barcoding methods, this approach does not require antibodies to surface markers, allowing for the labeling of all cells regardless of surface antigen expression. Additionally, since selenium and tellurium isotopes are not currently utilized in CyTOF antibody panels, this method expands barcoding options and frees up commonly used isotopes for more detailed cell profiling.

Efficient Neutralizing Antibodies Induction by Human Parvovirus B19 Epitope-Presenting Protein Nanoparticles.

Human parvovirus B19 (B19V) causes fetal hydrops in pregnant women. Despite the significant impact of this virus, effective vaccines remain unclear. In this study, we successfully engineered B19V protein nanoparticles by fusing the N-terminal receptor-binding domain corresponding to 5-80 amino acids of VP1 with two distinct types of self-assembling protein nanoparticles. Gel filtration and electron microscopic analysis confirmed the spherical assembly of the antigen-fused nanoparticles. The purified nanoparticles are efficiently bound to the surface of UT7/Epo-S1 cells, which are semi-permissive hosts for B19V infection. Immunization of BALB/c mice with VP1u 5-80 nanoparticles elicited a robust production of B19V-specific IgG antibodies compared to single VP1u 5-80 peptides. Moreover, a neutralization assay using B19V derived from a blood donor sample revealed that antibodies from mice immunized with VP1u 5-80 nanoparticles exhibited stronger infection-neutralizing activity. These findings suggest that nanoparticle formation plays a crucial role in enhancing the immunogenicity of the B19V VP1u 5-80 amino acid peptide and that these nanoparticles could serve as promising vaccine candidates, effectively inducing immunity against B19V.

IMMU-57. METHYL-B-CYCLODEXTRIN PARTIALLY REVERSES THE IMMUNOSUPPRESSIVE EFFECTS OF GLIOBLASTOMA DERIVED EXTRACELLULAR VESICLES ON LYMPHOCYTES

Abstract Extracellular vesicles (EVs) derived from glioblastoma (GBM) have been demonstrated in multiple studies to contribute to tumorigenesis, invasion, and immunosuppression. Our prior results demonstrate that EVs can induce the differentiation of myeloid-derived suppressor cells (MDSCs), which in turn impair T cell activation and proliferation in vitro. Methyl-B-Cyclodextrin (MBCD), a cholesterol depleting agent, can interfere with lipid rafts, which are cholesterol rich microdomains on the cellular membrane that act as a docking mechanism for EV uptake. Herein we demonstrate that MBCD can inhibit EV uptake and partially reverse the immunosuppressive effects of EVs on both monocytes and T cells. METHODS EVs were isolated using stepwise ultracentrifugation from GBM cultures. GBM derived EVs were then added to extracted monocytes from donor samples with or without MBCD for 72 hours under hypoxic conditions. The EV exposed monocytes were then co-cultured with T lymphocytes and cell proliferation was measured via flow cytometry. RESULTS Exposure to GBM-derived EVs induced MDSC differentiation in monocytes by at least 50% while the addition of MBCD reduced MDSC levels similar to that of the control group. Moreover, MBCD exposure resulted in increased T cell proliferation in the presence of EV treated monocytes compared to untreated samples. CONCLUSION MBCD can be used as a therapeutic agent to reverse MDSC induction by GBM derived EVs and can partially rescue T cell activation and proliferation.

A prospective study to evaluate the clinical specificity of the cobas® MPX test kit for screening for HIV RNA, HCV RNA, and HBV DNA in blood donation samples using the cobas® 6800 system in HBV endemic areas

BackgroundNucleic acid testing (NAT) is widely used for screening blood donors for infectious diseases to enhance transfusion safety. Roche's advanced cobas® MPX assay detects human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) using the cobas® 6800/5800 Systems, based on real-time PCR technology, providing improved sensitivity. This study aims to evaluate the clinical sensitivity and specificity of the cobas® MPX assay and its effectiveness in identifying infected donors in HBV endemic areas, particularly those with occult HBV infection (OBI). Materials and methodsA total of 12,067 donor samples from the Dalian Blood Center (DLBC, northern China) were tested for HIV, HCV, and HBV using both the cobas® MPX assay on the cobas® 6800 system and the previous generation cobas® TaqScreen MPX test v2.0 on the cobas s 201 system as the reference method. Testing was conducted using individual-donation testing (IDT) and primary pool of six donations (PP6), following the manufacturer's instructions and the operational procedures of the instruments. Samples with inconsistent results underwent repeated confirmation tests. ResultsCobas® MPX demonstrated 100.00 % overall percent agreement (95 % CI, 99.22 %-100.00 %) for IDT and 99.89 % (95 % CI, 99.82 %-99.95 %) for PP6. Kappa coefficients were 1.0 for IDT and 0.76 for PP6. Cobas® MPX specificity was 100.00 % (95 % CI, 99.22 %-100.00 %) for IDT and 99.99 % (95 % CI, 99.94 %-100.00 %) for PP6. Sensitivity was 100.00 % (95 % CI, 2.50 %-100.00 %) for IDT and 86.67 % (95 % CI, 68.36 %-95.64 %) for PP6. A total of 12 HBV NAT-yield cases were detected by cobas® MPX. ConclusionCobas® MPX demonstrated outstanding sensitivity and specificity in screening HIV, HCV, and HBV in routine blood donations, particularly enhancing occult HBV detection in endemic regions.