Unrelated Healthy Donors Research Articles

Shorter donor leukocyte telomere length is associated with poor peripheral blood stem cell mobilization induced by granulocyte colony-stimulating factor

Background/purposeInsufficient numbers of peripheral blood stem cells (PBSC) after granulocyte colony-stimulating factor (G-CSF) mobilization occurs in a significant proportion of PBSC collections, often from older age donors. Telomere length (TL) is often used as an indicator of an individual's biological age. This study aimed to investigate the relationship between donors' leukocyte TL and the outcome of G–CSF–induced PBSC mobilization in healthy unrelated donors. MethodsDonors’ leukocyte TLs and the outcome of G–CSF–induced PBSC mobilization, as assessed by pre-harvest CD34+ cell counts, were analyzed in 39 healthy PBSC donors. TL in a non-mobilized general population (n = 90) was included as a control group. G-CSF mobilization effect was categorized into three groups according to pre-harvest CD34+ cell count: poor (≤25/μL, PMD), intermediate (between 25 and 180/μL), and good (≥180/μl, GMD). ResultsLeukocyte TL of PBSC donors correlated well with pre-harvest CD34+ cell counts (r = 0.645, p < 0.001). Leukocyte TLs of PMDs (n = 8) were significantly shorter than those of GMDs (n = 9) and non-mobilization controls (p < 0.05). Moreover, all PMD TLs were below the 50th percentile, and 62.5% of PMDs had TLs below the 10th percentile of age-matched control participants. In contrast, no GMD TLs were below the 10th percentile; in fact, 33.3% (3/9) of them were above the 90th percentile. ConclusionOur results indicate that shorter donor leukocyte TL is associated with poor G–CSF–induced PBSC mobilization. TL, which represents a donor's biological age, could be a potential predictor for mobilization outcome.

Contribution of HLA class I (A, B, C) and HLA class II (DRB1, DQA1, DQB1) alleles and haplotypes in exploring ethnic origin of central Tunisians

BackgroundEstimation of HLA (Human leukocyte Antigen) alleles’ frequencies in populations is essential to explore their ethnic origin. Anthropologic studies of central Tunisian population were rarely reported. Then, in this work, we aimed to explore the origin of central Tunisian population using HLA alleles and haplotypes frequencies.MethodsHLA class I (A, B, C) and HLA class II (DRB1, DQA1, DQB1) loci genotyping of 272 healthy unrelated organ donors was performed by Polymerase Chain Reaction-Sequence Specific Oligonucleotide (PCR-SSO). We compared central Tunisians with other populations (Arabs, Berbers, Mediterraneans, Europeans, Africans, etc.) using alleles and haplotypes frequencies, genetic distances, Neighbour-Joining dendrogram and correspondence analysis.ResultsAmong the 19 HLA A alleles, the 26 HLA B alleles, the 13 HLA C alleles, the 15 HLA DRB1 alleles, the 6 HLA DQA1 alleles and the 5 HLA DQB1 alleles identified in the studied population, HLA A*02 (22.8%), HLA B*50 (13.1%), HLA C*06 (21.8%), HLA DRB1*07 (17.8%), HLA DQA1*01 (32.1%) and HLA DQB1*03 (31.6%) were the most frequent alleles. The extended haplotypes HLA A*02-B*50-C*06-DRB1*07-DQA1*02-DQB1*02 (1.97%) was the most frequent HLA six-loci haplotype.ConclusionCentral Tunisians were very close to other Tunisian populations, to Iberians and North Africans. They were rather distant from sub-Saharan populations and eastern Mediterraneans especially Arabs although the strong cultural and religious impact of Arabs in this population.

286. The Effect of Fecal Microbiota Transplantation on Decolonization of Multidrug-resistant Organisms in the gut

Abstract Background Gut colonization of multidrug-resistant organisms (MDROs), such as carbapenemase-producing Enterobacteriaceae (CPE) and vancomycin-resistant Enterococci (VRE), precedes clinical infections. It increases the risk of patient-to-patient transmission of MDROs as well as clinical infections such as bacteremia. Therefore, decolonization of MDROs in the gut by restoring colonization resistance through fecal microbiota transplantation (FMT) could be an effective infection control strategy for MDROs. Methods From March 2019 to November 2022, FMT for CPE and VRE decolonization in the gut was conducted at Hallym University Chuncheon Sacred Heart Hospital. Frozen or capsulized stool (MicroBiotix FMT Delivery Microbiota Preparation) from unrelated pre-screened healthy donor was used for each FMT. The rates of and the time to successful decolonization, defined as 3 consecutive negative results from rectal swab culture repeated with 3-day intervals after the initial FMT, were investigated in CPE and VRE rectal colonizers, respectively. Results FMT was performed in 60 CPE rectal carriers (KPC=55, KPC &amp; NDM=2, KPC &amp; IMP=1, NDM=1, NDM &amp; OXA=1) and in 15 VRE rectal carriers (E. faecium=14, E. faecalis=1). Among patients with follow-up data after initial FMT, decolonization success rates for CPE and VRE rectal carriers were 44.8% (26/58) and 66.7% (10/15) at 1 month, 77.1% (37/48) and 92.9% (13/14) at 3 months, 95.5% (42/44) and 100% (14/14) at 6 months, respectively. Median and mean times to decolonization after initial FMT were 30 and 43 days in CPE rectal colonizers and 14 and 29 days in VRE rectal colonizers, respectively. FMT results for decolonization of MDROs in the gut of Chuncheon Sacred Heart Hospital (ClinicalTrials.gov number, NCT04583098) Conclusion FMT as a microbiome restoration therapy is an effective method to control CPE and VRE colonization (ClinicalTrials.gov number, NCT04583098). Disclosures All Authors: No reported disclosures

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.

Efficacy and Safety of Fecal Microbiota Transplantation in the Treatment of Chronic Steroid-Refractory Graft-Versus-Host Disease after Hematopoietic Stem Cell Transplantation

Efficacy and safety of fecal microbiota transplantation in the treatment of chronic steroid-refractory graft-versus-host disease after hematopoietic stem cell transplantation Introduction The majority of patients who undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) with acute gut graft-versus-host disease has lost the diversity of gastrointestinal microbiota and associated to adverse clinical outcome. Despite glucocorticoids treatment has been considered as the first-line therapy regardless acute or chronic GVHD, the curative efficiency and durable response remains unsatisfactory. Several studies suggesting that fecal microbiota transplantation (FMT) from healthy donor effectively alleviated gut steroid-refractory acute GVHD (SR-aGVHD) by restoring diversity of gastrointestinal microbiota, resistance to potential pathogens, and transforming essential materials to metabolites for immunomodulating donor T cell which improved clinical outcomes in allo-HSCT. However, it is still unclear whether FMT therapy have similar potential efficacy in treating steroid-refractory chronic GVHD (SR-cGVHD). Here, we report the results of a pilot study from our single center using FMT from unrelated healthy donor to treat patients with SR-cGVHD. Method 12 patients aged 16-60 years with SR-cGVHD after allo-HSCT were enrolled. A total of 31 FMTs were performed via colonoscopy in 12 patients with SR-GvHD over three consecutive months. Adverse events with reasonable suspicion for being associated with FMT were monitored after treatment for 28 days. Partial response was defined as a decrease at least one stage in severity of GVHD or able to taper steroid dose. No response was defined as unable to taper steroid dose or need to increase or add immunosuppressor for controlling GVHD progression. CR was defined as the complete resolution of GVHD manifestations in all organs. To characterize the impact of the FMT on the gut microbiota, stool samples from recipients were collected the day before FMT treatment and 4 weeks after FMT treatment then carried out 16s rRNA sequencing. Result The overall response rate (ORR) in the patients with SR-cGvHD reached 50% (6/12), including CR in 0/0 (0%) cases and PR in 6/12 (50%). Most common adverse events were 3 abdominal bloating (9.68%), 2 diarrhea cases (6.45%) and 1 fever (3.23%). No serious adverse events were observed. The diversity of gastrointestinal microbiota remarkable decreased during SR-GVHD and it recovered after FMT (Figure B). After FMT treatment, similar α diversity of gastrointestinal microbiota in PR and NR patients was observed, whereas β-diversity analysis showed significant differences in the composition and abundance of the bacterial colonization between groups (Figure C and D). Conclusion The present study indicates that FMT as a new potential therapy in the treatment of SR-cGvHD is safe, well tolerated, and effective. Disclosures No relevant conflicts of interest to declare.

Association of &lt;I&gt;TNFAIP3&lt;/I&gt; (rs10499194) and &lt;I&gt;TNF-α&lt;/I&gt; (rs1800629) gene polymorphisms with susceptibility to systemic lupus erythematosus with juvenile onset and its clinical phenotypes in the Russian pediatric population

Numerous recent studies have shown that TNFAIP3 and TNF-α gene polymorphisms are associated with susceptibility to certain autoimmune and inflammatory diseases, including systemic lupus erythematosus (SLE), systemic scleroderma, rheumatoid arthritis, psoriasis, etc. However, the results of studies on associations between these polymorphisms and the risk of developing SLE in children are ambiguous and few in number.Objective: to test the hypothesis of a possible association between the rs10499194 polymorphism of the TNFA1P3 gene and the rs1800629 polymorphism of the TNF-α gene with susceptibility to juvenile SLE (jSLE) and its clinical phenotypes in the Russian pediatric population.Material and methods. Both polymorphisms were studied by allele-specific real-time polymerase chain reaction in 63 children (15 boys and 48 girls) with a confirmed diagnosis of jSLE, whose mean age was 12.3±3.2 years (3–17 years), and the mean duration of the disease was 4.1±2.4 years. Data on the frequency of genotypes and alleles of the corresponding TNFA1P3 and TNF-α gene polymorphisms in 309 healthy unrelated blood donors over the age of 18 years (20–45 years) were used as controls.Results and discussion. The study showed that the frequency of the rs10499194T mutant allele of the TNFA1P3 gene in patients with jSLE was significantly lower compared to the control (20.6 and 30.7%; p=0.023), and its carriage slightly reduced the risk of developing SLE (odds ratio, OR 0.58; 95% confidence interval, CI 0.32–1.05, p=0.053). The frequency of the rs1800629A mutant allele of the TNF-α gene was slightly higher in jSLE compared with controls (38.1 and 26.2%, respectively; p=0.056), and its carriage slightly increased the risk of developing SLE (OR 1.73; 95% CI 0.93–3.16; p=0.056). An analysis of the frequency distribution of the rs10499194 genotypes in groups of patients with and without arthritis revealed significant differences (p=0.003). Carrying genotypes with the mutant T allele (CT+TT genotypes) in jSLE significantly reduced the risk of developing of arthritis (p=0.003). At the same time, the risk of arthritis in carriers of at least one C allele was 3.76 times higher than in carriers of the other allele (p=0.006). No relationship was found between the rs1800629 TNF-α gene polymorphism and the clinical phenotypes of jSLE.Conclusion. The rs10499194T mutant allele statistically significant reduces the risk of arthritis development as one of the clinical manifestations of jSLE, and the rs1800629A mutant allele of the TNF-α gene is associated with a tendency to increase the risk of jSLE.

The recurrent pathogenic Pro890Leu substitution in CLTC causes a generalized defect in synaptic transmission in Caenorhabditis elegans.

De novo CLTC mutations underlie a spectrum of early-onset neurodevelopmental phenotypes having developmental delay/intellectual disability (ID), epilepsy, and movement disorders (MD) as major clinical features. CLTC encodes the widely expressed heavy polypeptide of clathrin, a major component of the coated vesicles mediating endocytosis, intracellular trafficking, and synaptic vesicle recycling. The underlying pathogenic mechanism is largely unknown. Here, we assessed the functional impact of the recurrent c.2669C > T (p.P890L) substitution, which is associated with a relatively mild ID/MD phenotype. Primary fibroblasts endogenously expressing the mutated protein show reduced transferrin uptake compared to fibroblast lines obtained from three unrelated healthy donors, suggesting defective clathrin-mediated endocytosis. In vitro studies also reveal a block in cell cycle transition from G0/G1 to the S phase in patient's cells compared to control cells. To demonstrate the causative role of the p.P890L substitution, the pathogenic missense change was introduced at the orthologous position of the Caenorhabditis elegans gene, chc-1 (p.P892L), via CRISPR/Cas9. The resulting homozygous gene-edited strain displays resistance to aldicarb and hypersensitivity to PTZ, indicating defective release of acetylcholine and GABA by ventral cord motor neurons. Consistently, mutant animals show synaptic vesicle depletion at the sublateral nerve cords, and slightly defective dopamine signaling, highlighting a generalized deficit in synaptic transmission. This defective release of neurotransmitters is associated with their secondary accumulation at the presynaptic membrane. Automated analysis of C. elegans locomotion indicates that chc-1 mutants move slower than their isogenic controls and display defective synaptic plasticity. Phenotypic profiling of chc-1 (+/P892L) heterozygous animals and transgenic overexpression experiments document a mild dominant-negative behavior for the mutant allele. Finally, a more severe phenotype resembling that of chc-1 null mutants is observed in animals harboring the c.3146 T > C substitution (p.L1049P), homologs of the pathogenic c.3140 T > C (p.L1047P) change associated with a severe epileptic phenotype. Overall, our findings provide novel insights into disease mechanisms and genotype-phenotype correlations of CLTC-related disorders.

Development of a calculated panel reactive antibody calculator for the United Arab Emirates: a proof of concept study

Calculated panel reactive antibody (CPRA) is used to help increase sensitized patient’s access to transplantation. United Arab Emirates (UAE) has a diverse resident population hence we developed a UAE–CPRA calculator based on HLA antigen frequencies of the different ethnic groups that represent the UAE population. HLA antigen frequencies at serological split antigen level for HLA-A, -B, -C, -DRB1 and -DQB1 of 1002 healthy unrelated donors were performed. We subsequently compared the performance of the UAE CPRA calculator with the Organ Procurement and Transplantation Network (OPTN) and the Canadian CPRA calculators in 110 Kidney Transplant waitlist patients from January 2016 to December 2018. Lin’s concordance correlation coefficient showed a moderate agreement between the UAE and OPTN calculator (Rc = 0.949, 95% CI 0.929–0.963) and the UAE and Canadian calculators (Rc = 0.952, 95% CI 0.932–0.965). While there continued to be a moderate agreement (Rc = 0.937, UAE versus OPTN calculator) in the lower sensitized group, a poor agreement (Rc = 0.555, UAE versus OPTN calculator) was observed in the higher sensitized group. In this study, we provide a template for countries to develop their own population-specific CPRA calculator. Implementation of the CPRA algorithm based on HLA frequencies of the multi-ethnic UAE population will be more fitting to increase access to transplantation and improve transplant outcomes. Our study demonstrates that the CPRA calculators developed using the data from the western population had poor correlation in our higher sensitized patients disadvantaging them in potential organ allocations systems. We plan to further refine this calculator by using high resolution HLA typing to address the problem of a genetically diverse population.

Genotyping Approach to Predict Coa and Cob Antigens in Thai Blood Donor Populations.

Coa and Cob antigens of the Colton (CO) blood group system are implicated in acute and delayed hemolytic transfusion reactions (HTRs). Owing to the inadequate supply of specific antiserum, data on CO phenotypes remain limited. This study aimed to develop genotyping methods to predict Coa and Cob antigens and to estimate transfusion-induced alloimmunization risks in three Thai blood donor populations. The study included 2451 blood samples from unrelated healthy Thai blood donors obtained from central, northern, and southern Thailand. DNA sequencing was used to determine the CO*A and CO*B alleles. In-house PCR with sequence-specific primers (PCR-SSP) and high-resolution melting curve (HRM) assays were performed and genotyping results were compared using DNA sequencing. CO*A and CO*B allele frequencies among Thais were determined using PCR-SSP and their frequencies were compared with other populations. The risks of Coa and Cob transfusion-induced alloimmunization among Thai donor populations were calculated. The validated genotyping results by PCR-SSP and HRM assays agreed with DNA sequencing. The CO*A/CO*A was the most common (100.0, 100.0, and 99.3%), followed by CO*A/CO*B (0.0, 0.0, and 0.7%) among central, northern and southern Thais. Homozygous CO*B/CO*B was not found. The CO*A and CO*B allele frequencies among central Thais significantly differed compared among southern Thais (p < 0.01) but not among northern Thais. Those allele frequencies among Thais were similar to those of Taiwanese, Chinese and Malay-Malaysian populations but not to South Asian, Southeast Asian, Korean, Japanese, Filipino, French Basque, and Maltese populations (p < 0.01). A higher risk of anti-Cob production rather than anti-Coa production was particularly noted in the southern Thai population. This study constitutes the first to determine CO*A and CO*B genotypes using PCR-SSP and HRM assays among Thais and this finding would be beneficial in predicting alloimmunization risk and providing safe transfusions among Thais.

Lysosomal alterations and decreased electrophysiological activity in CLN3 disease patient-derived cortical neurons.

CLN3 disease is a lysosomal storage disorder associated with fatal neurodegeneration that is caused by mutations in CLN3, with most affected individuals carrying at least one allele with a 966 bp deletion. Using CRISPR/Cas9, we corrected the 966 bp deletion mutation in human induced pluripotent stem cells (iPSCs) of a compound heterozygous patient (CLN3 Δ 966 bp and E295K). We differentiated these isogenic iPSCs, and iPSCs from an unrelated healthy control donor, to neurons and identified disease-related changes relating to protein synthesis, trafficking and degradation, and in neuronal activity, which were not apparent in CLN3-corrected or healthy control neurons. CLN3 neurons showed numerous membrane-bound vacuoles containing diverse storage material and hyperglycosylation of the lysosomal LAMP1 protein. Proteomic analysis showed increase in lysosomal-related proteins and many ribosomal subunit proteins in CLN3 neurons, accompanied by downregulation of proteins related to axon guidance and endocytosis. CLN3 neurons also had lower electrophysical activity as recorded using microelectrode arrays. These data implicate inter-related pathways in protein homeostasis and neurite arborization as contributing to CLN3 disease, and which could be potential targets for therapy.

Clinical Pharmacology and Determinants of Response to UCART19, an Allogeneic Anti-CD19 CAR-T Cell Product, in Adult B-cell Acute Lymphoblastic Leukemia.

UCART19 is an "off-the-shelf" genome-edited anti-CD19 chimeric antigen receptor (CAR)-T cell product, manufactured from unrelated healthy donor cells. UCART19 was administered to 25 adult patients with relapsed or refractory (R/R) B-cell acute lymphoblastic leukemia (B-ALL) in the CALM trial. All patients underwent lymphodepletion with fludarabine and cyclophosphamide ± alemtuzumab and received one of three ascending doses of UCART19. Given the allogeneic nature of UCART19, we analyzed the impact of lymphodepletion, HLA disparities, and host immune system reconstitution on its kinetics, along with other factors known to affect autologous CAR-T cell clinical pharmacology. Responder patients (12/25) had higher UCART19 expansion (C max) and exposure (AUCTlast) than nonresponders (13/25), as measured by transgene levels in peripheral blood. The persistence of CAR+ T cells did not exceed 28 days in 10/25 patients and lasted beyond 42 days in 4/25. No significant correlation was found between UCART19 kinetics and administered cell dose, patient and product characteristics or HLA disparities. However, the number of prior lines of therapy and absence of alemtuzumab negatively impacted UCART19 expansion and persistence. Alemtuzumab exposure positively affected IL7 and UCART19 kinetics, while negatively correlating with host T lymphocyte AUC0-28. UCART19 expansion is a driver of response in adult patients with R/R B-ALL. These results shed light on the factors associated with UCART19 kinetics, which remain highly affected by the impact of alemtuzumab on IL7 and host-versus-graft rejection. First description of the clinical pharmacology of a genome-edited allogeneic anti-CD19 CAR-T cell product showing the crucial role of an alemtuzumab-based regimen in sustaining UCART19 expansion and persistence through increased IL7 availability and decreased host T lymphocyte population.

Identification of cystic fibrosis transmembrane conductance regulator as a prognostic marker for juvenile myelomonocytic leukemia via the whole-genome bisulfite sequencing of monozygotic twins and data mining.

BackgroundLinked deoxyribonucleic acid (DNA) hypermethylation investigations of promoter methylation levels of candidate genes may help to increase the progressiveness and mortality rates of juvenile myelomonocytic leukemia (JMML), which is a unique myelodysplastic/myeloproliferative neoplasm caused by excessive monocyte and granulocyte proliferation in infancy/early childhood. However, the roles of hypermethylation in this malignant disease are uncertain.MethodsBone marrow samples from a JMML patient and peripheral blood samples from a healthy monozygotic twin and an unrelated healthy donor were collected with the informed consent of the participant’s parents. Whole-genome bisulfite sequencing (WGBS) was then performed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to analyze specific differentially methylated region (DMG) related genes. The target genes were screened with Cytoscape and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), which are gene/protein interaction databases. A data mining platform was used to examine the expression level data of the healthy control and JMML patient tissues in Gene Expression Omnibus data sets, and a survival analysis was performed for all the JMML patients.ResultsThe STRING analysis revealed that the red node [i.e., the cystic fibrosis transmembrane conductance regulator (CFTR)] was the gene of interest. The gene-expression microarray data set analysis suggested that the CFTR expression levels did not differ significantly between the JMML patients and healthy controls (P=0.81). A statistically significant difference was observed in the CFTR promoter methylation level but not in the CFTR gene body methylation level. The overall survival analysis demonstrated that a high level of CFTR expression was associated with a worse survival rate in patients with JMML (P=0.039).ConclusionsCFTR promoter hypermethylation may be a novel biomarker for the diagnosis, monitoring of disease progression, and prognosis of JMML.

Early fecal microbiome transfer after donor defecation determines response in patients with moderate to severe ulcerative colitis.

Fecal microbiome transfer (FMT) targeting gut microbiome dysbiosis is an emerging therapy for ulcerative colitis (UC). There is however no consensus on protocols for performing FMT in UC, especially in relation to time after donor feces defecation. This is a single-center retrospective analysis of patients with moderate-severe UC (total Mayo Clinic score ≥6 and endoscopic Mayo Clinic subscore of ≥2) treated with FMT between September 2017 and December 2019 at Dayanand Medical College and Hospital, Ludhiana,India. Fresh fecal samples from unrelated healthy voluntary donors were administered through colonoscopy at weeks 0, 2, 6, 10, 14, 18, and 22. Time interval between donor feces defecation and FMT procedure was recorded for each FMT session and the mean time of seven sessions was designated aika. Impact of aika on clinical response and safety of FMT was evaluated. During the study period, 123 adult patients (mean age 33.75±11.97 years, 61.8% [n=76] males) with moderate-severe UC (mean total Mayo Clinic and endoscopic Mayo Clinic scores 7.49±1.60 and 2.50±0.50, respectively) were treated with FMT. The mean aika was 2.29±0.75 h. The aika was smaller in patients who responded to FMT as compared to non-responders (2.13±0.75h vs. 2.71±0.76 h, p=0.0002) as well as in patients achieving clinical remission (2.15±0.76 h vs. 2.42±0.76 h, p=0.05). There was no significant impact of aika on adverse effects except forthe incidence of borborygmi after FMT, which was higher in patients with aika ≤2 h. Early FMT after donor feces defecation favorably impacts the clinical response rates in patients with active UC.

ABO, secretor, and Lewis carbohydrate histo-blood groups are associated with autoimmune neutropenia of early childhood in Danish patients.

BackgroundAutoimmune neutropenia of early childhood (AIN) is caused by autoantibodies directed against antigens on the neutrophil membrane. The ABO, secretor, and Lewis histo‐blood group systems control the expression of carbohydrate antigens and have previously been linked to autoimmune diseases. We aimed to investigate the association between genotypes and the risk of AIN in Danish patients.Study Design and MethodsOne hundred fifty‐four antibody‐positive AIN patients were included. Controls (n = 400) were healthy unrelated Danish blood donors. Molecular determination of ABO, secretor (FUT2), and Lewis (FUT3) genotypes were determined using real‐time polymerase chain reaction (qPCR) or Sanger sequencing to infer the prevalence of Lewis antigens (Lea and Leb) and secretor (SeSe or Sese) or nonsecretor (sese) phenotypes.ResultsBlood type O was more common in controls (46.8%) than in AIN patients (36.4%) (OR = 0.65; p = 0.028). Secretors of H Leb antigens were less frequent among AIN patients (25.2%) than controls (35.0%) (OR = 0.62; p = 0.037).DiscussionABO blood group antigens and the secretion of these antigens are associated with a diagnosis of AIN. The mechanism underlying the association between autoimmunity and interaction among ABO, secretor, and Lewis genotypes has not yet been elucidated, but several studies indicate a connection to the gut microbiota.

Multi-Donor Fecal Microbial Transplantation for Critically Ill Patients: Rationale and Standard Operating Procedure

Patients in the intensive care unit often lose a considerable fraction of their gut microbiome due to exposure to broad-spectrum antibiotics and other reasons. Dysbiosis often results in prolonged diarrhea and increase occurrence of multi-drug resistant pathogens in the colon with clinical consequences not yet well understood. Restoring the microbiome by fecal microbial transplantation (FMT) is a plausible therapeutic possibility, so far only documented in case reports and case series using very heterogeneous methodologies. Before FMT with critically ill patients can be tested in randomized controlled trials, there is a burning need to describe a standardized operating procedure (SOP) of the whole process, respecting the specifics of the critically ill population, such as the risk of the disrupted intestinal barrier and time-critical nature of the procedure. We describe the SOP that has been developed for experimental use in critically ill patients by a multidisciplinary team of intensivists, gastroenterologists, and microbiologists based on feedback from regulatory authority (State Institute for Drug Control of the Czech Republic). The hallmarks of these SOPs are multi-donor freshly frozen transplants guaranteed for 2 months consisting of seven aliquots from seven unrelated healthy donors and administered by a rectal tube. In this paper we discuss the rationale for this SOP and the process of its development in detail and release the full proposed SOP is in the form of an online appendix.

RHCE*E and RHCE*e genotype incompatibility in a southern Thai Muslim population.

CONTEXT:The formation of red cell alloantibodies resulting from both transfusion and pregnancy can cause adverse effects from allogeneic blood transfusions. Alloanti-E is commonly detected among Thai and Asian populations.AIMS:This study aimed to determine RHCE*E and RHCE*e genotype incompatibility in a southern Thai Muslim population and to compare it with those previously reported for other populations.SUBJECTS AND METHODS:Nine hundred and twenty-seven DNA samples obtained from 427 unrelated healthy blood donors from southern Thai Muslims and 500 samples from Central Thais were included. Samples were genotyped for RHCE*E and RHCE*e using an in-house polymerase chain reaction with the sequence-specific primer technique.RESULTS:Significant differences were found when we compared the allele frequencies of the RHCE*E and RHCE*e between southern Thai Muslims and Central Thais: RHCE*E 0.162 versus 0.197 and RHCE*e 0.838 versus 0.803 and also found in Chinese, American native, Japanese, Korean, Alaskan native, Hawaiian, South Asian, Brazilian Japanese-descendant, and Malay Malaysian populations (P < 0.05). In addition, the E/e incompatibilities among southern Thai Muslims and Central Thais were 24.23% and 26.71%, respectively.CONCLUSIONS:This study was the first to determine the RHCE*E and RHCE*e genotype incompatibility among southern Thai Muslims, enabling the estimation of their potential alloimmunization risk. These data could be useful to provide safe blood transfusions across ethnic populations.

Generation of human induced pluripotent stem cell lines from four unrelated healthy control donors carrying European genetic background

Four human induced pluripotent stem cell (hiPSC) lines have been generated from healthy control European donors, and validated. This resource represents a useful tool for stem cell-based research, as references for developmental studies and disease modeling linked to any type of human tissue and organ, in an ethnical-, sex- and age-matched context. They providea reliable in-vitro model for single cell- and tissue-based investigations, and are also a valuable tool for genome editing-based studies.

A Pilot Study of MG4101, Allogeneic Natural Killer Cell, in Patients with Relapsed or Refractory Acute Myeloid Leukemia

Background: Prognosis of relapsed or refractory acute myleoid leukemia (AML) is dismal. Several studies demonstrated that allogeneic natural killer (NK) cells have potent anti-tumor effect in AML. In this study, we aimed to investigate the safety and efficacy of allogeneic, ex vivo expanded killer-cell immunoglobulin-like receptor (KIR) B haplotype NK cells (MG4101) in refractory or relapsed AML and to elucidate the association between immunologic characteristics and clinical response.Methods: Patients were scheduled to receive MG4101 for a total of 2 cycles for 28 days in each cycle. After lymphodepletion with fludarabine and cyclophosphamide, MG4101 derived from unrelated healthy donors was administered intravenously for 3 days during the first 3 weeks of the treatment cycle followed by IL-2 for 3 days. Throughout the entire treatment schedule, it was aimed to deliver a total of 2.0 - 5.0 x 10 9 cells to each patient based on actual body weight. NK cell receptor ligand (NKRL) in AML blast and KIR typing of donors and recipients were evaluated and analyzed with the clinical response of the patients.Results: Between Apr 2018 to Feb 2020, a total of 11 patients (male, n=5) with refractory or relapsed AML patients were conesecutively enrolled. 5 patients (45.5%) were intermediate risk group, and 6 patients (54.5%) were adverse risk group. Three patients had antecedent hematologic malignancies (atypical chronic myeloid leukemia [n=1], myelodysplastic syndrome [n=2]). Of 7 evaluable patients, 2 patients (28.6%) showed partial response (PR) and 2 patients (28.6%) showed stable disease (SD). Median overall survival of entire patients was 3.4 months (95% confidence interval, 2.5 - 4.3 months): 3.7 months in responders (PR or SD) and 1.0 months in nonresponders. There was only 1 grade 4 infusion-related hypersensitivity reaction, which resolved with supportive care, and there was no cytokine release syndrome, or any sign of graft-versus-host disease associated with MG4101 treatment.NKRL expression varied widely among the patients treated with MG4101. In patients with favorable clinical outcome, ULBP3, activating NKRL for NKG2D receptor, was highly expressed or increased with treatment. PD-L1 expression, in contrast, was maintained relatively low until the end of the treatment. Besides, activating KIR and its cognate ligand interaction were associated with treatment response. Among the responders including PR and SD, the sum of activating KIR tended to be higher than non-responders.Conclusions: MG4101 in refractory or relapsed AML was well tolerated without significant treatment-related toxicity. NKRL expression in AML blasts was heterogeneous, and treatment response with MG4101 was also highly variable. In our patient cohort, patients with high activating KIR responded favorably to MG4101 treatment. These findings suggests further investigation of MG4101 in patients in relapsed or refractory AML and highlights the importance of immunologic mechanism in treating myeloid neoplasm. [Display omitted] DisclosuresKoh: Pfizer: Consultancy; Jassen: Honoraria; AstraZeneca: Honoraria; Novartis: Honoraria; GSK: Honoraria; Roche: Honoraria; Takeda: Honoraria. Kim: Seoul National University Hospital: Current Employment.

The relation between clinical and objective gait scores in clubfoot patients with and without a relapse

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with complex genetic inheritance. Many reports have provided evidence that signal transducer and activator of transcription-4 (STAT4) may participate in the pathogenesis of SLE.The aim was to investigate the clinical significance and possible association of STAT4 (G/T) genetic polymorphism and the susceptibility to SLE in a cohort of Egyptian female patients.Sixty-five Egyptian SLE female patients and 100 age and sex-matched unrelated female healthy blood donors who served as controls, were included in the study. STAT4 genotyping was performed by real time PCR-allelic discrimination technique.STAT4 genotyping in patients revealed that 63.1% had GG, 32.3% GT and 6.15% wild (TT) genotype. There was a non-significant difference in the distribution of STAT4 genotypes between patients and controls. Vasculitis, photosensitivity and lupus nephritis were significantly increased in patients with the homomutant (GG and TT) compared to heteromutant (GT) genotype (p = 0.01, p = 0.04 and p < 0.01 respectively). Patients with a TT genotype had a significantly consumed C3 and C4 levels and higher anti-dsDNA positivity compared to those with GG and GT genotypes. Promoter polymorphism tended to be higher in juvenile-onset SLE cases.STAT4 (G/T) polymorphism was not associated with an increased risk of SLE in Egyptian females. However, vasculitis, photosensitivity and renal involvement were significantly higher among patients harboring the homomutant genotypes. Genetic polymorphism may be an important determinant affecting disease progression and is associated with DNA positivity and younger age of onset.

Shorter Interdonation Interval Contributes to Lower Cell Counts in Subsequent Stem Cell Donations

Approximately 7% of unrelated hematopoietic stem cell donors are asked to donate stem cells a subsequent time to the same or a different recipient. Recent studies have shown that donation-related symptoms for second donations are similar to those for the first donation. Little is known about differences in stem cell mobilization and yields for subsequent peripheral blood stem cell (PBSC) and bone marrow (BM) collections. We hypothesized that CD34+ cell yields and total nucleated cell (TNC) concentrations for subsequent PBSC or BM donations are lower than those at the first donation. We also evaluated the factors influencing stem cell yields in healthy unrelated second-time donors. Data were gathered from the Center for International Blood and Marrow Transplant Research database on 513 PBSC and 43 BM donors who donated a second time between 2006 and 2017 through the National Marrow Donor Program. Among the second-time PBSC donors, we found significantly lower preapheresis peripheral blood CD34+ cell counts (68.6×106/L versus 73.9×106/L; P=.03), and collection yields (556×106 versus 608×106; P=.02) at the second donation compared to the first. This decrease at the subsequent donation was associated with a shorter interdonation interval, lower body mass index (BMI), and a lower total G-CSF dose. In most instances, suboptimal mobilizers at their first donation donated suboptimal numbers of stem cells at their subsequent donations. Among repeat BM donors, the TNC concentration was lower at the second donation. The small size of this group precluded additional analysis. Overall, when considering repeat donations, increasing the interdonation intervals and evaluating for BMI changes should be considered to optimize stem cell yields. Some of these parameters may be improved by increasing G-CSF dose in PBSC donors within permissible limits.