Donor Cells Research Articles

The health risk of social disadvantage is transplantable into a new host

Low socioeconomic status (SES) is a risk factor for mortality and immune dysfunction across a wide range of diseases, including cancer. However, cancer is distinct in the use of allogeneic hematopoietic cell transplantation (HCT) as a treatment for hematologic malignancies to transfer healthy hematopoietic cells from one person to another. This raises the question of whether social disadvantage of an HCT cell donor, as assessed by low SES, might impact the subsequent health outcomes of the HCT recipient. To evaluate the cellular transplantability of SES-associated health risk, we analyzed the health outcomes of 2,005 HCT recipients who were transplanted for hematologic malignancy at 125 United States transplant centers and tested whether their outcomes differed as a function of their cell donor's SES (controlling for other known HCT-related risk factors). Recipients transplanted with cells from donors in the lowest quartile of SES experienced a 9.7% reduction in overall survival (P = 0.001) and 6.6% increase in treatment-related mortality within 3 y (P = 0.008) compared to those transplanted from donors in the highest SES quartile. These results are consistent with previous research linking socioeconomic disadvantage to altered immune cell function and hematopoiesis, and they reveal an unanticipated persistence of those effects after cells are transferred into a new host environment. These SES-related disparities in health outcomes underscore the need to map the biological mechanisms involved in the social determinants of health and develop interventions to block those effects and enhance the health of both HCT donors and recipients.

Ethical Considerations in Nonconsenting Adult Allogeneic Stem Cell Donors.

Ethical Considerations in Nonconsenting Adult Allogeneic Stem Cell Donors.

Quality Control in Human Adipose-Derived Stromal/Stem Cells and Tissue Engineering Fat Models for Aging Studies.

Adipose tissue is recognized not only as an endocrine organ but also as a reservoir for adipose-derived stromal/stem cells (ASCs). ASCs have stimulated the interest of both the scientific and medical communities due to their therapeutic potential and applications in tissue engineering and regenerative medicine. ASCs are leveraged for their multipotency and their paracrine function. ASC behavior is highly variable and donor dependent. Donor age, body mass index, disease status, sex, and ethnicity can lead to differential overall function and quality. The impact of donor age and passage on ASC behavior has been well documented, impacting cell proliferation and differentiation potential and thus must be taken into careful consideration when conducting in vitro studies. Pooling of ASCs from different donors reduces heterogeneity among individual donors and produces ASCs with a consistent differentiation and paracrine profiles, an advantage for studies in biological aging. This chapter provides a detailed overview for studies related to quality control for ASC pools considering biological and chronological aging in ASCs. There are hallmarks of biological aging and specific assays associated with the evaluation of each hallmark. Nevertheless, here we present the assays that provide a standardized characterization and qualification of donor pools for their regenerative potential, considering chronological and biological age of the pool. The assays included in this chapter are considered quality control standards to evaluate cell proliferation, differentiation, colony-forming units, and cellular senescence from different donor age and cell passage cohorts.

Research Progress in the Establishment of Sterile Hosts and Their Usage in Conservation of Poultry Genetic Resources

Many local chicken breeds are rapidly declining and even facing extinction due to a variety of factors such as indiscriminate crossbreeding, climate fluctuations, epidemic outbreak, and environmental changes. Developing effective preservation strategies is important to address this situation. The special reproductive characteristics and gamete morphology of chickens pose challenges for preserving genetic heritage through the cryopreservation of genetic materials. Currently, gonad and primordial germ-cell cryopreservation and transplantation are the most promising approaches, especially for the genetic information in the W chromosome of female birds. The study of establishing sterile recipients is crucial for increasing the efficiency of the colonization of transplanted donor tissues and cells. Several classical methods, including ovariectomy and testectomy, busulfan, and irradiation, have been employed to deplete endogenous germ cells in recipient males before transplantation. These methods rely on the toxicity of chemical reagents and physical stimulation to kill germ cells. Recent advances in gene-editing technology have introduced sterile hosts via the knocking out of genes relevant to germ cells’ development. This review explores state-of-the-art technologies for preparing infertile avian recipients (mainly chickens) and aims to provide guidance for the conservation of poultry genetic material and breed restoration.

The ethical and legal challenges of cell donation for brain organoid research

The ethical and legal challenges of cell donation for brain organoid research Ethical and legal concerns raised by the use of human biological materials, especially cells from adult donors and foetal tissues, must be thoroughly examined. This article delves into these challenges, with a specific focus on the issues of consent in cell donation for brain organoid research. Brain organoid research represents a significant leap forward in neuroscience and offers unprecedented insights into brain development and disorders. However, the use of human biological materials, particularly cells from adult donors and foetal tissues, raises critical ethical and legal concerns. (6, 10) This report examines these challenges by focusing on the issues of consent in the donation of cells for brain organoid research.

Bioengineered Abiotic Nanomaterials Through Cell Membrane-Camouflaging: Advancements and Challenges in Lung Cancer.

Lung cancer remains a major global health concern with high mortality rates and poor prognosis. Bridging the gap between the chemical and cellular understanding of cell-decorated biomimetic nanocomposites and their clinical translation is crucial for developing effective therapies. Nanocomposites show promise in targeted drug delivery and diagnostics, but their clinical application is hindered by biocompatibility and clearance issues. To overcome these challenges, biomimetic approaches utilizing cell membrane-coated nanomaterials emerge. By camouflaging nanomaterials with cell membranes, the biointerfaces are enhanced, and the inherent properties of the donor cell membranes are acquired. This review provides an overview of recent advancements on cell membrane-coated nanocomposites for lung cancer diagnosis and treatment. It discusses fabrication techniques, biomedical applications, challenges, and future prospects. The incorporation of cell membranes into nanocomposites holds potential for improved lung cancer therapy, but further development and refinement are needed for precise tumor targeting. Addressing the identified challenges will pave the way for clinical translation of these biomimetic nanoplatforms and advance lung cancer diagnosis and treatment.

Cryo-EM structure of the R388 plasmid conjugative pilus reveals a helical polymer characterized by an unusual pilin/phospholipid binary complex.

Bacterial conjugation is a process by which DNA is transferred unidirectionally from a donor cell to a recipient cell. It is the main means by which antibiotic resistance genes spread among bacterial populations. It is crucially dependent upon the elaboration of an extracellular appendage, termed "pilus," by a large double-membrane-spanning secretion system termed conjugative "type IV secretion system." Here we present the structure of the conjugative pilus encoded by the R388 plasmid. We demonstrate that, as opposed to all conjugative pili produced so far for cryoelectron microscopy (cryo-EM) structure determination, the conjugative pilus encoded by the R388 plasmid is greatly stimulated by the presence of recipient cells. Comparison of its cryo-EM structure with existing conjugative pilus structures highlights a number of important differences between the R388 pilus structure and that of its homologs, the most prominent being the highly distinctive conformation of its bound lipid.

Pharmacokinetics of Briquilimab as a Conditioning Agent for Hematopoietic Stem Cell Transplantation in Patients with Severe Combined Immunodeficiency, Myelodysplastic Syndrome, or Acute Myeloid Leukemia

BackgroundFor successful engraftment of donor hematopoietic stem cells (HSC), conditioning with chemotherapy and/or radiation prior to hematopoietic cell transplantation (HCT) has been required to open marrow niche space and minimize the risk of immune rejection. Briquilimab, a humanized IgG1 monoclonal antibody that blocks the interaction between the c-Kit receptor and stem cell factor on various C-Kit expressing tissues including HSC, is a potential nonmyeloablative conditioning agent in clinical development for patients with severe combined immunodeficiency (SCID), myelodysplastic syndromes (MDS), and acute myeloid leukemia (AML). Objective(s)This study aimed to characterize pharmacokinetics (PK) and develop a population PK model of briquilimab after single intravenous infusions of 4 different doses in patients with SCID, MDS, or AML receiving HCT. Study DesignThe PK data was collected from 2 different studies: JAS-BMT-CP-001 and JSP-CP-003. JAS-BMT-CP-001 is a phase 1/2 open-label study of briquilimab as a conditioning agent prior to allogenic HCT in SCID patients. The participants received single intravenous infusions of 0.1, 0.3, 0.6, or 1.0 mg/kg. JSP-CP-003 was a phase 1a/b open-label study of briquilimab in combination with a standard conditioning regimen of low dose total body irradiation and fludarabine in MDS or AML subjects undergoing HCT. The participants received a single intravenous dose of 0.6 mg/kg briquilimab. In both studies, briquilimab PK samples were obtained at pre-treatment, 5 minutes post-end of infusion, 4- and 24-hours post-start of infusion, any time between 2 days and 30 days post-infusion, and on the day of HCT prior to donor cell infusion.The population PK model was developed using the PK data from these 2 clinical studies, and the effect of participants’ baseline characteristics on the briquilimab PK was evaluated. PK simulations were performed using the developed PK model to calculate the time to reach target concentrations for HCT. ResultsA total of 49 participants (21 SCID adult and pediatric participants with a median age of 12 years and 28 MDS/AML adult participants with a median age of 70 years) were included in the PK analysis. A two-compartment model with combined linear and non-linear elimination best described the PK of briquilimab. Body weight was determined as the sole covariate of the PK parameters among the explored covariates. For a typical subject with a body weight 70 kg, the estimated parameters for clearance, maximum metabolic rate of Michaelis Menten elimination, Michaelis Menten constant, central volume, peripheral volume, and intercompartmental clearance were 17.6 mL/hr, 51434.8 ng/hr, 71.5 ng/mL, 3444.0 mL, 1613.3 mL, and 21.2 mL/hr, respectively. The median time to reach target concentrations of 500, 1000, and 2000 ng/mL after a single dose of 0.6 mg/kg was calculated as 12.3, 10.4, and 7.7 days, respectively. ConclusionsThe PK of intravenous briquilimab was characterized in subjects with SCID, MDS, or AML receiving HCT, and a population PK model was developed to estimate briquilimab clearance to use as a guide to the timing of donor cell infusion post-briquilimab. Body weight was identified as a significant covariate on elimination and volume of distribution of briquilimab.

Quantifying and Controlling the Proteolytic Degradation of Cell Adhesion Peptides.

Peptides are widely used within biomaterials to improve cell adhesion, incorporate bioactive ligands, and enable cell-mediated degradation of the matrix. While many of the peptides incorporated into biomaterials are intended to be present throughout the life of the material, their stability is not typically quantified during culture. In this work, we designed a series of peptide libraries containing four different N-terminal peptide functionalizations and three C-terminal functionalizations to better understand how simple modifications can be used to reduce the nonspecific degradation of peptides. We tested these libraries with three cell types commonly used in biomaterials research, including mesenchymal stem/stromal cells (hMSCs), endothelial cells, and macrophages, and quantified how these cell types nonspecifically degraded peptides as a function of terminal amino acid and chemistry. We found that peptides in solution which contained N-terminal amines were almost entirely degraded by 48 h, irrespective of the terminal amino acid, and that degradation occurred even at high peptide concentrations. Peptides with C-terminal carboxylic acids also had significant degradation when cultured with the cells. We found that simple modifications to the termini could significantly reduce or completely abolish nonspecific degradation when soluble peptides were added to cells cultured on tissue culture plastic or within hydrogel matrices, and that functionalizations which mimicked peptide conjugations to hydrogel matrices significantly slowed nonspecific degradation. We also found that there were minimal differences in peptide degradation across cell donors and that sequences mimicking different peptides commonly used to functionalize biomaterials all had significant nonspecific degradation. Finally, we saw that there was a positive trend between RGD stability and hMSC spreading within hydrogels, indicating that improving the stability of peptides within biomaterial matrices may improve the performance of engineered matrices.

Study on the Construction of Family Support System for Unrelated Hematopoietic Stem Cell Donation Volunteers

There are millions of patients in urgent need of hematopoietic stem cell transplantation in China, but the matching success rate of unrelated hematopoietic stem cells is only 1/400 ~ 1/10000. Since the establishment of China Bone Marrow Bank in 1992, only more than 14,000 patients have been successfully matched. According to the survey, a large number of volunteers in China will eventually choose to refuse to donate hematopoietic stem cells, which is closely related to the public's lack of understanding of hematopoietic stem cell donation. Unrelated volunteers can get less support from family members than related hematopoietic stem cell donors, which leads to their lower donation rate. Therefore, it is of great practical significance to study the family support system of unrelated hematopoietic stem cell donation volunteers.

P-796 A novel method of somatic cell nuclear transfer to generate artificial oocytes

Abstract Study question Can we identify an alternative method to Sendai-Virus fusion for somatic cell nuclear transfer (SCNT) aiming at generating artificial oocyte? Summary answer The utilization lipofectamine proved itself as a valid alternative method to virus-mediated SCNT, allowing successful membrane fusion and supporting proper embryo development. What is known already SCNT involves transferring a diploid cell’s nucleus into an enucleated oocyte. Early attempts of SCNT involves direct injection and electrofusion but both approaches can affect oolemma integrity. To avoid mechanical damage to oolemma, fusogenic agents have been employed for SCNT including polyethylene glycol (PEG) and more efficient Hemagglutinating Virus of Japan Envelope (HVJ-E). However, despite its high efficiency, HVJ-E is difficult to be used in human because it is a virus derivative, and a recent shortage of this compound made it not readily available. On the other hand, cationic phospholipid is fusogenic and is widely utilized in transfection experiments. Study design, size, duration In the past 4 months, SCNT was carried out through PEG-mediated, HVJ-E-mediated, or novel lipofectamine-mediated fusion using cumulus cells as nuclear donor. Post-SCNT oocytes were inseminated and cultured up to the blastocyst stage. Embryo development and morphokinetics were assessed and compared among the three SCNT experimental cohorts. In some cases, blastocysts were transferred into pseudo-pregnant mice, post-natal development were monitored. Fertilization, embryo development, and offspring wellbeing were monitored. Participants/materials, setting, methods Oocytes from B6D2F1 mice were enucleated by micromanipulation. The ooplasts were allocated to 3 cohorts: 1) exposure to 45% PEG, 2) subzonal transfer of donor cell with HVJ-E, or 3) subzonal transfer the donor cell exposed to 1.75% lipofectamine. Reconstructed oocytes were monitored for fusion 30 minutes after exposure to fusogens and inseminated by spermatozoa from B6-EGFP mice to confirm paternal genome provenance. Fertilization and embryo development were monitored in a time-lapse system. Main results and the role of chance A total of 165 oocytes were enucleated and 160 (97.0%) survived and allocated to PEG-fusion (n = 50), Sendai-virus-fusion (n = 41) and lipofecamine-fusion (n = 80) cohorts. Fusion rate was 56.0% (28/50) for the PEG-fusion cohort, while Sendai-virus and lipofectamine-fusion cohorts yielded higher fusion rate at 92.7%(38/41) and 86.3%(69/80), respectively (P<0.0001). Control oocytes yielded 96.7% (145/150) fertilization rate and developed to 2-cell stage at 93.8% (136/145), and yielded an eventual blastocyst rate at 80.7% (117/145). Despite a lower oocyte reconstruction rate, PEG-fusion cohort fertilized at 89.3% (25/28), but cleavage rate to 2-cell stage was lower at 48.0% (12/25, P<0.0001) and did not yield any blastocyst. The HVJ-E cohort yielded a fertilization rate of 94.7% (36/38) and cleavage rate at 97.2% (35/36) similar to control. The blastocyst rate was observed at 22.9% (8/35, P<0.0001) when comparing to control. Lipofectamine-fusion cohort yielded 91.3% (63/69) fertilization rate and subsequent cleavage rate at 95.2% (60/63). Albeit at a lower blastocyst development rate comparing to control, the lipofectamine cohort yielded similar full-preimplantation rate at 19.0% (12/63) when comparing to HVJ-E cohort. All blastocysts (n = 20) were transferred into 2 pseudo-pregnant mice and 3 healthy female offspring were yielded to date. Limitations, reasons for caution HVJ-E mediated membrane fusion has been well established with proven safety record; however, its recent shortage has led to a temporary cessation of its utilization. Our lipofectamine-mediated cell fusion technique yielded satisfactory fusion, fertilization, blastocyst formation and some live offspring, however the safety and efficacy still remain to be validated. Wider implications of the findings This novel lipofectamine-mediated cell fusion has broader implications for advancing cloning technologies. Lipofection is a non-viral approach, overcomes shortage of HVJ-E and therefore propose itself as a more appealing cell fusion method for human SCNT. Trial registration number N/A

P-798 A feasible clinical model to generate genotyped oocytes for infertile women

Abstract Study question Is it possible to generate genotyped oocytes by utilizing accessible somatic cells in a somatic cell nuclear transfer model? Summary answer Genotyped oocytes generated through haploidization of endometrial somatic cells exhibited reliably completed embryo development and yielded healthy offspring. What is known already The generation of genotyped oocytes through neogametogensis or somatic cell nuclear transfer (SCNT) would be the ultimate solution to treat female infertility due to primary ovarian insufficiency or agonadism. Among the various approaches, somatic cell haploidization appears more feasible (Lee, et al 2022). Unfortunately, this approach has only been successful in a murine model using cumulus cells. However, this donor cell type is not ideal when translated into clinical setting, due to the absence of cumulus cells in patients who are void of oocytes, which arguably is the patient population that would potentially benefit the most from this technique. Study design, size, duration In a mouse SCNT model, endometrial stromal cells (EMSCs) were injected into enucleated oocytes. Cumulus cells (CCs) were used as another source of nuclear donor. Intact MII oocytes were used as control. Post-SCNT oocytes and controls were inseminated by spermatozoa from B6-EGFP mice to monitor paternal genome presence through embryo development and offspring. Embryos were cultured in a time-lapse microscope. To prove gamete competence, blastocysts were transferred into pseudo-pregnant recipients and offspring health was assessed. Participants/materials, setting, methods Endometrial stromal cells were cultured for 4 days and exposed to aphidicolin overnight to synchronize at G1 phase of cell cycle. Cumulus cells were collected after denudation. Oocytes from B6D2F1 mice were enucleated under polarized light microscope. SCNT was performed with CCs or EMSCs by lipofectamine-mediate fusion. Oocytes were inseminated with spermatozoa from GFP-positive transgenic mice. Embryos morphokinesis were assessed by a time-lapse system. Blastocysts were transferred into 2.5-dpc surrogate CD1 mice. Main results and the role of chance Control oocytes(n = 75) fertilized at 93.3%(70/75) and progressed to 2-cell stage at 93.3%(70/75), 8-cell at 93.3%(70/75) and blastocyst at 89.3%(67/75). The control embryos started syngamy at 14.4±1h, reaching the 4-cell stage at 37.3±3h and blastulation at 80.6±8h. A total of 484 oocytes were enucleated, 470(97.1%) survived and underwent SCNT using CCs(n = 85) or EMSCs(n = 385). Fusion rate was 95.5% for CC with a haploidization rate of 70.9%(56/79) that led to a fertilization rate of 64.3%(36/56). The CC cohort developed to 2-cell, 8-cell, and morula at 47.4%, 35.4%, and 22.8%, with a delay in development at syngamy at 19.0±8h(P<0.0001). In the EMSC cohort, fusion rate was 96.0%(361/376). Haploidization rate was 65.4%(236/361), comparable to the CC cohort(P = 0.35). Fertilization was lower in the EMSC cohort at 47.0%(P<0.01) compared to the CC cohort. Subsequently, EMSC cohort yielded similar cleavage rates to 2-cell stage at 30.7%, 8-cell stage at 29.4%, and morula stage at 28.5%. The blastocyst rate was higher in the EMSC cohort at 28.0%(P<0.05). The EMSC cohort yielded similar developmental timing comparing to controls from pronuclear appearance to 4-cell cleavage but a delay manifested afterward at 37.3±3h(P<0.0001). In the EMSC cohort, 42 blastocysts were transferred, yielding 3 healthy pups(2 male,1 female). Limitations, reasons for caution While we generated consistent haploidization and satisfactory fertilization, full pre-implantation development remained low. Even more limited was the live birth rate, which probably related to subtle factors such as an imprinting issue due to insufficient reprogramming of the somatic cell donor. Wider implications of the findings The SCNT model described to generate genotyped oocytes, although still with limited efficiency, represents a feasible reproductive option for women with agonadism and compromised ovarian reserve. This model is still a better approach to complete neogametogenesis. Trial registration number not applicable

Serum from patients with cirrhosis undergoing liver transplantation induces permeability in human pulmonary microvascular endothelial cells ex vivo

IntroductionPatients with cirrhosis undergoing liver transplantation frequently exhibit systemic inflammation, coagulation derangements, and edema, indicating endothelial dysfunction. This syndrome may worsen after ischemia–reperfusion injury of the liver graft, coincident with organ dysfunction that worsens patient outcomes. Little is known about changes in endothelial permeability during liver transplantation. We hypothesized that sera from these patients would increase permeability in cultured human endothelial cells ex vivo.MethodsAdults with cirrhosis presenting for liver transplantation provided consent for blood collection during surgery. Sera were prepared at five time points spanning the entire operation. The barrier function of human pulmonary microvascular endothelial cells in culture was assessed by transendothelial resistance measured using the ECIS ZΘ system. Confluent cells from two different endothelial cell donors were stimulated with human serum from liver transplant patients. Pooled serum from healthy men and purified inflammatory agonists served as controls. The permeability response to serum was quantified as the area under the normalized resistance curve. Responses were compared between time points and analyzed for associations with clinical characteristics of liver transplant patients and their grafts.ResultsLiver transplant sera from all time points during surgery-induced permeability in both endothelial cell lines. The magnitude of permeability change was heterogeneous between patients, and there were differences in the effects of sera on the two endothelial cell lines. In one of the cell lines, the severity of liver disease was associated with greater permeability at the start of surgery. In the same cell line, serum collected 15 min after liver reperfusion induced significantly more permeability as compared to that collected at the start of surgery. Early postreperfusion sera from patients undergoing living donor transplants induced more permeability than sera from deceased donor transplants. Sera from two exemplary cases of patients on preoperative dialysis, and one patient with an unexpectedly long warm ischemia time of the liver graft, induced exaggerated and prolonged endothelial permeability.DiscussionSerum from patients with cirrhosis undergoing liver transplantation induces permeability of cultured human pulmonary microvascular endothelial cells. Increased endothelial permeability during liver transplantation may contribute to organ injury and present a target for future therapeutics.

Dynamics of DNA methylation during osteogenic differentiation of porcine synovial membrane mesenchymal stem cells from two metabolically distinct breeds

ABSTRACT Mesenchymal stem cells (MSCs), with the ability to differentiate into osteoblasts, adipocytes, or chondrocytes, show evidence that the donor cell’s metabolic type influences the osteogenic process. Limited knowledge exists on DNA methylation changes during osteogenic differentiation and the impact of diverse donor genetic backgrounds on MSC differentiation. In this study, synovial membrane mesenchymal stem cells (SMSCs) from two pig breeds (Angeln Saddleback, AS; German Landrace, DL) with distinct metabolic phenotypes were isolated, and the methylation pattern of SMSCs during osteogenic induction was investigated. Results showed that most differentially methylated regions (DMRs) were hypomethylated in osteogenic-induced SMSC group. These DMRs were enriched with genes of different osteogenic signalling pathways at different time points including Wnt, ECM, TGFB and BMP signalling pathways. AS pigs consistently exhibited a higher number of hypermethylated DMRs than DL pigs, particularly during the peak of osteogenesis (day 21). Predicting transcription factor motifs in regions of DMRs linked to osteogenic processes and donor breeds revealed influential motifs, including KLF1, NFATC3, ZNF148, ASCL1, FOXI1, and KLF5. These findings contribute to understanding the pattern of methylation changes promoting osteogenic differentiation, emphasizing the substantial role of donor the metabolic type and epigenetic memory of different donors on SMSC differentiation.

Genetic ablation of adhesion ligands mitigates rejection of allogeneic cellular immunotherapies.

Allogeneic cellular immunotherapies hold promise for broad clinical implementation but face limitations due to potential rejection of donor cells by the host immune system. Silencing of beta-2 microglobulin (B2M) expression is commonly employed to evade Tcell-mediated rejection by the host, although the absence of B2M is expected to trigger missing-self responses by host natural killer (NK) cells. Here, we demonstrate that genetic deletion of the adhesion ligands CD54 and CD58 in B2M-deficient chimeric antigen receptor (CAR) Tcells and multi-edited induced pluripotent stem cell (iPSC)-derived CAR NK cells reduces their susceptibility to rejection by host NK cells invitro and invivo. The absence of adhesion ligands limits rejection in a unidirectional manner in B2M-deficient and B2M-sufficient settings without affecting the antitumor functionality of the engineered donor cells. Thus, these data suggest that genetic ablation of adhesion ligands effectively alleviates rejection by host immune cells, facilitating the implementation of universal immunotherapy.

Thymus gland and thymic dysfunctions in children

The thymus is the organ that has gained significant awareness in the last two centuries, being considered the main donor of cells for the lymphatic system, much like the bone marrow is the donor of cells for the cardiovascular system. Currently, knowledge regarding the thymus gland has considerably improved. Thymic dysfunctions encompass congenital and acquired conditions, which manifest through organ hypoplasia or hyperplasia, as well as formations/tumors of the anterior mediastinum.

The impact of consanguinity on the design of iPSC banks.

The effect of consanguinity on identifying universal induced pluripotent stem cell (iPSC) donors, i.e., homozygous for the major human leukocyte antigen (HLA) loci, is unknown. The discovery sample size was calculated in a consanguineous population using a method (1qF) based on the inbreeding coefficient. The result was orders of magnitude smaller compared to the standard method.

IN VITRO STUDY OF THE DOSE-DEPENDENT EFFECT OF JANUS KINASE INHIBITORS

Background. One of the most promising means of targeting the mechanisms of autoimmune processes and hyperimmune reactions associated with cytokine imbalance are Janus kinase inhibitors. However, these chemicals in the used doses have a number of undesirable side effects. The effect of small and ultra-low doses in relation to this group of substances has not been studied. Purpose. To study the effect of low and ultra-low concentrations of Janus kinase inhibitors on the synthesis of cytokines by human mononuclear cells in vitro. Materials and methods. The concentrations of tumor necrosis factor alpha (TNFα), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon alpha (IFNα), interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 8 (IL-8) and interleukin 10 (IL-10) after stimulation with phytohemagglutinin were determined in the culture of mononuclear cells of healthy donors. Also the effect on the production of these cytokines of ruxolitinib and baricitinib in various doses, including ultra-low ones, was determined. Results. It was found that baricitinib, as well as ruxolitinib, at normal concentrations significantly decreased, and at ultra-low concentrations they significantly increased the level of synthesis of all cytokines studied in this experiment. Conclusion. The stimulating, but not suppressive effect of ultra-low concentrations of Janus kinase inhibitors has been demonstrated using the example of ruxolitinib and baricitinib. In relation to inhibitors of Janus kinase functions, this phenomenon has been discovered for the first time, which opens up ways to search for new means of regulating immunity, in particular, the production of cytokines.

Islet-antigen reactive B cells display a unique phenotype and BCR repertoire in autoantibody positive and recent-onset type 1 diabetes patients.

Autoreactive B cells play an important but ill-defined role in autoimmune type 1 diabetes (T1D). To better understand their contribution, we performed single cell gene and BCR-seq analysis on pancreatic islet antigen-reactive (IAR) B cells from the peripheral blood of nondiabetic (ND), autoantibody positive prediabetic (AAB), and recent-onset T1D individuals. We found that the frequency of IAR B cells was increased in AAB and T1D. IAR B cells from these donors had altered expression of B cell signaling, pro-inflammatory, infection, and antigen processing and presentation genes. Both AAB and T1D donors demonstrated a significant increase in certain heavy and light chain V genes, and these V genes were enriched in islet-reactivity. Public clones of IAR B cells were restricted almost entirely to AAB and T1D donors. IAR B cells were clonally expanded in the autoimmune donors, particularly the AAB group. Notably, a substantial fraction of IAR B cells in AAB and T1D donors appeared to be polyreactive, which was corroborated by analysis of recombinant monoclonal antibodies. These results expand our understanding of autoreactive B cell activation during T1D and identify unique BCR repertoire changes that may serve as biomarkers for increased disease risk.

Hemoglobin Electrophoresis versus Kleihauer-Betke to Determine Bone Marrow Suppression in Fetuses Undergoing Intrauterine Transfusion.

Mainstay therapy for fetuses affected by maternal red cell alloimmunization is serial intrauterine transfusion (IUT). Testing to determine when fetal red cells have been replaced with donor cells historically involves the use of the Kleihauer-Betke (KB) test. Hemoglobin (Hgb) electrophoresis testing may be more rapid with a reduced cost of analysis. We aimed to determine the correlation between fetal Hgb electrophoresis versus the traditional KB test. This is a retrospective analysis of all alloimmunized singleton pregnancies undergoing IUT between January 1, 2021, and July 1, 2023. Maternal and fetal characteristics were collected along with the indication for IUT. A final fetal blood sample was obtained at the conclusion of each transfusion and sent for KB testing and Hgb electrophoresis. The primary outcome was the assessment of these parameters in their ability to predict the replacement of the fetal circulating red cell population with donor cells. Linear regression analysis and repeated measures analysis of variance were performed, and p-values less than 0.05 were considered significant. A total of 56 IUTs were performed in 16 patients. There were 39 (69.6%) final KB test values collected and compared with 30 (53.6%) final Hgb electrophoresis values. Hgb electrophoresis when compared with the KB test demonstrated a significant correlation (R 2 = 0.93; 95% confidence interval, 0.61-0.76; p < 0.001). This same finding held true when examining the correlation at each individual IUT as well. The final KB test and Hgb electrophoresis values significantly decreased with each transfusion (p = 0.003). A predominance of adult donor blood was noted by the third transfusion for both laboratory indices. Fetal Hgb electrophoresis obtained at the time of IUT demonstrates a significant correlation with the traditional KB test. · Fetal Hgb electrophoresis following IUT is underexplored. · Hgb electrophoresis is an automated evaluation. · The traditional KB test is a manual evaluation. · These two tests demonstrate significant correlation.