Differential gene expression in regulatory T-cells after activation by temporary contact with dendritic cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE3996 | GEO | 2007/01/09
SECONDARY ACCESSION(S): PRJNA94271
REPOSITORIES: GEO
Similar Datasets
Project description:Regulatory CD4+CD25+ T-cells were isolated from human blood using MACS and were then left untreated or were cocultured overnight with immature monocyte-derived Dendritic cells. Treg's total RNAs were prepared after moDC-removal. Experiment Overall Design: One sample per treatment-group. No replicates.
2008-06-12 | E-GEOD-3996 | biostudies-arrayexpress
Project description:Introduction: Expansion of antigen (Ag)-specific natural occurring regulatory T cells (nTregs) is required to obtain sufficient numbers of cells for cellular immunotherapy. In this study, different allogeneic stimuli were studied for their capacity to generate functional alloAg-specific nTregs. Methods: A highly enriched nTreg-fraction (CD4+CD25brightCD127- T cells) was alloAg-specific expanded using HLA-mismatched immature, mature monocyte-derived dendritic cells (moDC) or peripheral blood mononuclear cells (PBMC). The allogeneic mature moDC-expanded nTregs were fully characterized by analysis of the demethylation status within the TSDR of the FOXP3 gene and the expression of both protein and mRNA of FOXP3, HELIOS, CTLA4 and cytokines. In addition, the antigen-specific suppressive capacity of these expanded nTregs was tested. Results: Allogeneic mature moDC and skin-derived DC were superior in inducing nTreg-expansion compared to immature moDC or PBMC in an HLA-DR and CD80/CD86-dependent way. Remarkably, the presence of exogenous IL-15 without IL-2, could facilitate optimal mature moDC-induced nTreg-expansion. Allogeneic mature moDC-expanded nTregs were at low ratios (<1:320), potent suppressors of alloAg-induced proliferation without significant suppression of completely HLA-mismatched-Ag-induced proliferation. Mature moDC-expanded nTregs were highly demethylated at the TSDR within the FOXP3 gene and highly expressed of FOXP3, HELIOS and CTLA4. A minority of the expanded nTregs produced IL-10, IL-2, IFN-g and TNF-a but very few IL-17 producing nTregs were found. Next generation sequencing of mRNA of moDC-expanded nTregs revealed a strong induction of Treg-associated mRNAs. Conclusions: Human allogeneic mature moDC are highly efficient stimulator cells, in presence of exogenous IL-15, for expansion of stable alloAg-specific nTregs with superior suppressive function. Four different batches of highly pure regulatory T cells (all from the same donor) were expanded in two different ways, and compared to non-expanded samples.
2015-04-20 | E-GEOD-66385 | biostudies-arrayexpress
Project description:Introduction: Expansion of antigen (Ag)-specific natural occurring regulatory T cells (nTregs) is required to obtain sufficient numbers of cells for cellular immunotherapy. In this study, different allogeneic stimuli were studied for their capacity to generate functional alloAg-specific nTregs. Methods: A highly enriched nTreg-fraction (CD4+CD25brightCD127- T cells) was alloAg-specific expanded using HLA-mismatched immature, mature monocyte-derived dendritic cells (moDC) or peripheral blood mononuclear cells (PBMC). The allogeneic mature moDC-expanded nTregs were fully characterized by analysis of the demethylation status within the TSDR of the FOXP3 gene and the expression of both protein and mRNA of FOXP3, HELIOS, CTLA4 and cytokines. In addition, the antigen-specific suppressive capacity of these expanded nTregs was tested. Results: Allogeneic mature moDC and skin-derived DC were superior in inducing nTreg-expansion compared to immature moDC or PBMC in an HLA-DR and CD80/CD86-dependent way. Remarkably, the presence of exogenous IL-15 without IL-2, could facilitate optimal mature moDC-induced nTreg-expansion. Allogeneic mature moDC-expanded nTregs were at low ratios (<1:320), potent suppressors of alloAg-induced proliferation without significant suppression of completely HLA-mismatched-Ag-induced proliferation. Mature moDC-expanded nTregs were highly demethylated at the TSDR within the FOXP3 gene and highly expressed of FOXP3, HELIOS and CTLA4. A minority of the expanded nTregs produced IL-10, IL-2, IFN-g and TNF-a but very few IL-17 producing nTregs were found. Next generation sequencing of mRNA of moDC-expanded nTregs revealed a strong induction of Treg-associated mRNAs. Conclusions: Human allogeneic mature moDC are highly efficient stimulator cells, in presence of exogenous IL-15, for expansion of stable alloAg-specific nTregs with superior suppressive function.
2015-04-20 | GSE66385 | GEO
Project description:Modulating beta-catenin signaling has attractive therapeutic potential in cancer immunotherapy. Various studies have found that beta-catenin can mediate immune evasion in cancer and promote anti-inflammatory features of antigen-presenting dendritic cells. Multiple small-molecule compounds that inhibit Wnt/beta-catenin signaling are currently in clinical development, but none have reached routine clinical use. Thus, new inhibitors of beta-catenin signaling are desirable. This dataset is the result of an investigation of the effect of small molecular compounds axitinib, ICG-001, nitazoxanide, orlistat, and XAV-939 on the maturation capacity of monocyte-derived dendritic cells (moDC). Treatment of immature moDC was done in combination with the beta-catenin activator 6-BIO. Immature moDCs were obtained by isolating monocytes from PBMC using magnetic beads, which were then differentiated into moDCs using GM-CSF and IL-4 for four days. Compounds were added during the maturation of the moDCs, which was done by adding LPS to the cell culture. After the maturation period of 24 hours, the mature moDCs were collected and processed for RNA sequencing.
2021-08-06 | E-MTAB-10757 | biostudies-arrayexpress
Project description:Human monocyte-derived dendritic cells (moDCs) have been used as an in vitro model for studying tolerance and immunity. However, the underlying metabolic states of tolerogenic (dexamethasone and vitamin D3-treated), immature and immunogenic (mature, LPS-treated) moDCs have not been completely characterized. Through transcriptomic analyses, we determined that tolerogenic moDCs exhibit augmented catabolic pathways with respect to oxidative phosphorylation (OXPHOS), fatty acid oxidation (FAO) and glycolysis. Functionally, tolerogenic moDCs showed the highest mitochondrial membrane potential, production of reactive oxygen species and superoxide, and increased mitochondrial spare respiratory capacity. Tolerogenic and mature moDCs manifested differential FAO gene expression with FAO activity being significantly higher in tolerogenic and immature moDCs than in mature. In addition, tolerogenic and mature moDCs demonstrated similar levels of glycolytic rate, but not glycolytic capacity and reserve, which were more pronounced in tolerogenic and immature moDCs. Finally, tolerogenic and immature moDCs, but not mature moDCs, showed high plasticity to compensate the intracellular ATP content after inhibition of different energetic metabolic pathways. Overall, tolerogenic moDCs exhibit a metabolic signature of increased, stable OXPHOS programing and high plasticity for metabolic adaptation. These findings provide a framework for future research of metabolic properties of human DCs. Total RNA of sixteen samples from four moDC types (tolerogenic, LPS-tolerogenic, immature and mature) were extracted by Trizol (Invitrogen) followed by a clean-up procedure using RNeasy Micro Kit (Qiagen). All RNA samples had an integrity number ≥9.6 assessed by Agilent Bioanalyzer. Total RNA samples were amplified using TargetAmp™ and the biotinilated cRNA was prepared by Nano-g™ Biotin-aRNA Labeling Kit for the Illumina® System (Epicentre). After the hybridization to the Illumina Human HT-12 v4 Beadchips for 17 h at 58°C, the arrays were washed, stained (Illumina Wash Protocol) and then scanned using BeadArray Scanner 500GX. Array data were extracted at the probe level without background correction using Illumina GenomeStudio software. These raw data were quantile normalized and log2 transformed. Technical replicates were obtained from the hybridization in duplicate of three samples. Pearson correlation analysis showed high correlation between the technical replicates (r>0.99). Differentially expressed genes (DEGs) were identified using Limma16 with Benjamini-Hochberg multiple testing correction (p<0.05). DEGs were further clustered into different groups according to the patterns of expression change among the different moDC types using STEM software17. The analysis was performed in R v.2.12.2 (http://www.R-project.org) with Bioconductor 2.12 (http://www.bioconductor.org) and enabled by Pipeline Pilot (www.accelrys.com).
2015-05-08 | E-GEOD-52894 | biostudies-arrayexpress
Project description:Viruses have evolved numerous strategies to impair immunity so that they can replicate more efficiently, such as the immunosuppressive effects of Morbillivirus infection. In the present work we hypothesized that the highly contagious morbillivirus Peste des Petits Ruminants virus (PPRV) could target monocytes and dendritic cells (DC) to contribute to the immunosuppressive effects produced by the infection. Monocytes isolated from healthy sheep donor, a natural host of the disease, could be infected by PPRV and this impaired the differentiation and phagocytic ability of immature monocyte-derived DC (MoDC). Ovine MoDC could be productively infected by PPRV, and this drastically reduced MoDC capacity to activate allogeneic T cell responses. Transcriptomic analysis of infected MoDC indicated that several tolerogenic DC signature genes were upregulated upon PPRV infection.
2022-09-06 | GSE203515 | GEO
Project description:Purpose: The goals of this study are to compare NGS-derived Treg transcriptome profiling (RNA-seq) to microarray methods and to evaluate phenotypic difference between Tregs cocultured with control and Capns1-knockdown SVEC4-10 cells. Methods: Isolated splenic CD4+CD25+ Tregs were cocultured with SVEC4-10 cells for 5 days. Then total RNA in cocultured Tregs were isolated using an RNeasy mini kit (Qiagen) with on-column DNaseI digestion. Three replicates per each groups were pooled, and were subjected to the RNA-seq analysis using a NovaSeq6000 (Illumina, San Diego, CA) to obtain ~40 million 150 bp paired-end reads by Eurofins genomics (Tokyo, Japan). Contlusions: There is no remarkable difference in gene profiels between Tregs cocultured with the control and Capns1-knockdown SVEC4-10 cells.
2021-02-19 | GSE167072 | GEO
Project description:To investigate the gene expression of MoDC after PRRSV stimulation and nucleocapsid protein transfection
2016-08-30 | GSE86182 | GEO
Project description:Buffy coats from four independent human donors were enriched for CD4+ cells using MACS CD4 beads and LS columns. Cells were subsequently sorted into Naïve, Central Memory and Effector Memory using following markers: Naive: CD4+CD25–CD45RA+CCR7+; Central Memory: CD4+CD25–CD45RA–CCR7+; Effector Memory: CD4+CD25-CD45RA-CCR7-. Total RNA from these cell subsets was extracted and 100ng was used for Nanostring SPRINT run according to manufacturer's instructions. Overall aim of the experiment is assessing the expression level of human T helper cell miRNome.
2022-01-10 | GSE161098 | GEO
Project description:Allergic contact dermatitis (ACD) is the most prevalent form of immunotoxicity in humans. Dendritic cells (DC) play a central role in the pathogenesis of ACD, particularly during the sensitization phase to a specific allergen. Upon activation, DCs maturate and migrate to draining lymph nodes which is accompanied by major metabolic and phenotypic alterations that facilitate the presentation of the captured antigen to prime naïve T cells. Nevertheless, knowledge on the molecular effects during maturation of DCs in the context of ACD remain scarce. Here, we present a global proteomic analysis of monocyte-derived dendritic cells (MoDC) treated with NiSO4, the most prominent cause of ACD. Proteomic alterations induced after treatment with NiSO4 were compared to the bacterial trigger lipopolysaccharide (LPS). Both substances possess a similar TLR4 binding capacity, which may help to identify allergy-specific effects compared to bacterial activation. MoDCs treated for 24 h with 2.5 µg/ ml LPS displayed a very strong immunological response, characterized by upregulation of DC activation markers, secretion of proinflammatory cytokines and stimulation of T cell proliferation. Similar immunological responses were observed after treatment with 400 µM NiSO4 but less pronounced. Both triggered TLR4 and TREM1 pathways. However, NiSO4 in addition, also activated hypoxic and apoptotic pathways, which might have overshadowed initial signaling. Moreover, our proteomic data support the importance of Nrf2 as a key player in mediating sensitization since many Nrf2 targets genes were strongly upregulated on protein level selectively after treatment with NiSO4. Strikingly, NiSO4 stimulation induced cellular hypocholesterolemia which was counteracted by the induction of genes and proteins relevant for cholesterol biosynthesis. Our proteomic study allowed for the first time to better characterize some of the fundamental differences between NiSO4 and LPS-triggered activation of MoDCs, providing an important contribution to the molecular understanding of contact allergy.
2021-03-29 | PXD022599 | Pride