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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Functional Analysis and Gene Expression Profile of Umbilical Cord Blood Regulatory T Cells

ABSTRACT: Since the role of cord blood (CB) regulatory T cells (Tregs) for the suppression of the allogeneic T-cell response is under investigation, we analyzed and compared the functional properties and gene expression profile of Tregs expanded from CB units or from the peripheral blood (PB) of helathy donors. Tregs were purified from 23 CB units and from the PB of 13 donors and expanded for 6 days with anti-CD3, anti-CD28 and IL-2. Immunophenotypic analyses were performed, and suppressor activity of expanded Tregs was measured in mixed lymphocyte reaction (MLR) cultures. The IL-10 production capacity was tested and gene expression profile experiments were performed on 6 Tregs from PB and 4 from CB. CB and PB Tregs had similar immunophenotypic features. Tregs from CB presented a higher expansion capacity and genomic characterization showed in CB-derived Tregs a significant enrichments of genes involved in cell proliferation, chromatin modification and regulation of gene expression in CB-derived Tregs. All samples were positive for the Foxp3 gene and protein after expansion. CB and PB expanded Tregs exerted a comparable and potent suppressive function of MLR and presented a high in vitro IL-10 production capacity. Gene profile analysis also revealed for PB Tregs a significant enrichments of genes involved in the adaptive immune response. Comparison of the transcriptional profile of T-REGS derived either from cord blood or peripheral blood. 6 highly purified T-regs samples derived from peripheral blood of healthy donors were compared to highly purified T-regs from cord blood.

ORGANISM(S): Homo sapiens

SUBMITTER: Sabina Chiaretti

PROVIDER: E-GEOD-22501 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Functional Analysis and Gene Expression Profile of Umbilical Cord Blood Regulatory T Cells

Project description:Since the role of cord blood (CB) regulatory T cells (Tregs) for the suppression of the allogeneic T-cell response is under investigation, we analyzed and compared the functional properties and gene expression profile of Tregs expanded from CB units or from the peripheral blood (PB) of helathy donors. Tregs were purified from 23 CB units and from the PB of 13 donors and expanded for 6 days with anti-CD3, anti-CD28 and IL-2. Immunophenotypic analyses were performed, and suppressor activity of expanded Tregs was measured in mixed lymphocyte reaction (MLR) cultures. The IL-10 production capacity was tested and gene expression profile experiments were performed on 6 Tregs from PB and 4 from CB. CB and PB Tregs had similar immunophenotypic features. Tregs from CB presented a higher expansion capacity and genomic characterization showed in CB-derived Tregs a significant enrichments of genes involved in cell proliferation, chromatin modification and regulation of gene expression in CB-derived Tregs. All samples were positive for the Foxp3 gene and protein after expansion. CB and PB expanded Tregs exerted a comparable and potent suppressive function of MLR and presented a high in vitro IL-10 production capacity. Gene profile analysis also revealed for PB Tregs a significant enrichments of genes involved in the adaptive immune response.

2010-12-31 | GSE22501 | GEO

Human ECFCs: Cord Blood vs Peripehral Blood ECFCs

Project description:miRNA profiling was carried out using the miRCURY LNA™ microRNA Array (5th gen - hsa, mmu & rno). microRNA profiling of CB and PB-derived ECFCs from 3 independent donors from each cell source. To identify and understand the regulation of endothelial cell functions through comparing miRNA expression profiling of cord blood (CB) derived endothelial colony forming cells (ECFCs) and peripheral blood (PB) ECFCs

2017-03-20 | GSE81574 | GEO

Genetic module and miRNome trait analyses reflect the distinct biological features of endothelial progenitor cells from different anatomic locations

Project description:Background: Endothelial progenitor cells (EPCs) play a fundamental role in post-natal vascular repair, yet EPCs from different anatomic locations possess unique biological properties. The underlying mechanisms are unclear. Method: We performed transcriptome analysis for EPCs isolated from 2 different sources: cord blood (CB) or adult peripheral blood (PB). Both gene expression microarray and small RNA sequencing (smRNA-seq) technologies were applied. Results: EPCs from CB expressed abundant genes involved in cell cycle, hypoxia signalling and blood vessel development, correlating with the phenotypes that CB-EPCs proliferated more rapidly, migrated faster, and formed tubule structure more efficiently. smRNA-seq further deciphered miRNome patterns in EPCs isolated from CB or PB: 54 miRNAs were enriched in CB-EPCs, while another 50 in PB-EPCs. Specifically, CB-EPCs expressed more angiogenic miRNAs such as miR-31, while PB-EPCs possessed more tumor suppressive miRNAs including miR-10a. Knocking down miR-31 levels in CB-EPCs suppressed cell migration and microtubule formation, while overexpressing miR-31 in PB-EPCs helped to recapitulate some of CB-EPC functions. Conclusion: Our results show the foundation for a more detailed understanding of EPCs from different anatomic sources. Stimulating the expression of angiogenic microRNAs or genes in EPCs of low activity (such as those from patients with cardiovascular diseases) might allow the development of novel therapeutic strategies. EPC from cord blood or peripheral blood that outgrown after 2-4 week culture were collected. The RNA are extracted and profiled by Affymetrix GeneChip U133 plus 2.0 expression array. This submission represents gene expression microarray component of study.

2012-09-21 | E-GEOD-39763 | biostudies-arrayexpress

Genetic module and miRNome trait analyses reflect the distinct biological features of endothelial progenitor cells from different anatomic locations

2012-09-21 | GSE39763 | GEO

Nanostring CAR T Characterization panel comparing expression in cord blood-derived transgenic T cells cultured with IL-2 or with IL-7 and IL-15, peripheral blood-derived transgenic T cells cultured with IL-2

Project description:The purpose of the experiment was to transcriptionally characterize transgenic T cells derived from cord blood (CB) and cultured with IL-2 or IL-7 and IL-15, or from peripheral blood (PB) and cultured with IL-2. The sample replicates were generated using peripheral and cord blood mononuclear cells, obtained from healthy human donors and cord blood units that were not compliant with banking standards, respectively. T cells were then transduced with a retroviral vector encoding an HLA-DRB1*04-restricted TCR targeting the human papillomavirus oncoprotein E7.

2021-10-15 | GSE161483 | GEO

Comparative gene expression analyses of purified CD14+ monocytes from human umbilical cord blood, CD14+ monocytes from adult peripheral blood and the cell therapy product DUOC-01

Project description:DUOC-01 is a macrophage-like cell therapy product manufactured from human umbilical cord blood (CB) mononuclear cells (MNCs) and intended for use in treatment of demyelinating diseases. During the 21-day manufacturing process, most initiating MNCs die in culture, but a highly active, adherent population of cells that resemble hematopoietic lineage macrophage in morphology, phagocytic activity, and antigen expression emerges; this population comprises the final DUOC-01 product [Kurtzberg et al. 2015]. Using the cuprizone-mediated murine demyelination model, we have found that DUOC-01 cells are capable of accelerating the remyelination process. In the same demyelination model, human cord blood-derived CD14+ cells (CBCD14) were less active. In another cellular therapy project, we have found that CBCD14 protects brain cells from oxygen-glucose-deprivation (OGD), but adult peripheral blood-derived CD14+ monocytes (PBCD14) did not. To address the functional differences between these cell types, we have analyzed differential gene expression by these three cell types. We used four cord blood (CB) samples (from Carolinas Cord Blood Bank) that were collected within 24hrs of delivery. Then, highly purified CB CD14+ monocytes were isolated by flow sorting. We collected the peripheral blood (PB) from four healthy volunteers and isolated the purified PB CD14+ monocytes by cell sorting. Three separate batches of cultured DUOC-01 cells were collected from Robertson Cell and Translational Cell Therapy cGMP facility. Total RNAs were isolated at the same time and kept frozen. RNA samples were given to Duke Microarray core facility to analyse the gene expression profile using Affymetrix GeneChipÂ® Human Genome U133 Plus 2.0 Assay.

2016-01-13 | E-GEOD-76803 | biostudies-arrayexpress

regulatory T cells in human pregnancy

Project description:Transplacental immune programming refers to the concept that during pregnancy, significant crosstalk occurs between the maternal and fetal immune system, with consequent long‐term effects especially for the offspring. In this study, we searched for evidence of transplacental programming in cellular immunity. We made the surprising observation that there is a stringent and specific correlation of regulatory T cells (Treg) between the mother and the fetus. Gene microarray analysis indicates that interleukin 10 (IL‐10) might be involved in transplacental Treg alignment. This is further supported by the direct correlation of IL‐10 at a protein level between maternal fetal dyads. In vitro data provide evidence that IL‐10 may regulate the homeostatic balance between Tregs and non‐Tregs through selective upregulation of the anti‐apoptotic molecule Bcl‐2 in Tregs. Induction of IL‐10 might be triggered by fetus‐derived estriol (E3), which readily crosses the placenta and correlates with maternal IL‐ 10 levels. Our study represents the first example of transplacental regulation of cellular immunity between the mother and the fetus. These novel findings might have major implications for pregnancy related‐changes in the maternal and fetal immune system, and provide possible mechanistic insights into how prenatal influences can lead to subsequent immunopathologies, such as allergy and autoimmunity. T-reg and non-Treg samples of non pregnant women, and matched cord blood and maternal blood T-regs and non-T-regs

2013-11-04 | E-GEOD-31976 | biostudies-arrayexpress

Methylome of Fetal and Maternal Monocytes and Macrophages at the Feto-Maternal Interface

Project description:Maternal and fetal monocytes and tissue macrophages (decidual macrophages, Hofbauer cells) at the feto-maternal interface have different methylome. Paired and balanced design. We compared maternal blood monocytes (MB) vs. cord blood monocytes (CB), maternal blood monocytes (MB) vs. decidual macrophages (Deci), cord blood monocytes (CB) vs placental macrophages (villi) and decidual macrophages (Deci) vs. placental macrophages (villi).

2011-11-15 | E-GEOD-31680 | biostudies-arrayexpress

Expression data from cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, both while resting & after stimulation

Project description:Here we compare the effects of stimulation on cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, isolated by MACS protocols & expanded in vitro using dynabeads. Expansion was carried out at a ratio of 3 beads/cell in the presence of 1000units/ml of recombinant human IL2 for 8 days, followed by 3 days of culture without beads. RNA was extracted from resting cells on day 4 after expansion. Stimulated cells were treated with Ionomycin for 2hrs prior to RNA extraction. Four way comparison of resting and stimulated Tregs versus Thelpers. A total of 18 arrays were analysed containing RNA from 5 individual cord blood donors.

2010-06-22 | E-GEOD-20934 | biostudies-arrayexpress

p57Kip2 alterations in adult CFU-E contribute to steroid resistance in Diamond Blackfan anemia

Project description:Ashley R, Yan H, Wang N, Hale J, Dulmovits BM, Papoin J, Olive ME, Udeshi ND, Carr SA, Vlachosa A, Lipton JM, Da Costa L, Hillyer C, Kinet S, Taylor N, Mohandas N, Narla A, Blanc L. 2019. Despite the effective clinical use of steroids for the treatment of Diamond Blackfan anemia (DBA), the mechanistic bases via which glucocorticoids regulate human erythropoiesis remain poorly understood. Here, we report that the sensitivity of erythroid differentiation to dexamethasone (Dex) is dependent on the developmental origin of human CD34+ progenitor cells, specifically increasing the expansion of CD34+ progenitors from peripheral blood (PB) but not cord blood (CB). Dexamethasone treatment of erythroid-differentiated PB, but not CB, CD34+ progenitors resulted in the expansion of a novel CD34+CD36+CD71hiCD105med immature colony-forming unit-erythroid (CFU-E) population. Furthermore, proteomics analyses revealed the induction of distinct proteins in dexamethasone-treated PB and CB erythroid progenitors. Dexamethasone treatment of PB progenitors resulted in the specific upregulation of p57Kip2, a Cip/Kip cyclin-dependent kinase inhibitor, and we identified this induction as critical; shRNA-mediated downregulation of p57Kip2, but not the related p27Kip1, significantly attenuated the impact of dexamethasone on erythroid differentiation and inhibited the expansion of the immature CFU-E subset. Notably, in the context of DBA, we found that steroid resistance was associated with a dysregulated p57Kip2 expression. Altogether, these data identify a novel glucocorticoid-responsive human erythroid progenitor and provide new insights into glucocorticoid-based therapeutic strategies for the treatment of patients with DBA.

2019-11-23 | MSV000084614 | MassIVE

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