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Transcriptional profiling of human Th9 T cell clones treated with GW9662 against untreated control

ABSTRACT: Transcriptional profiling of human Th9 T cell clones treated with GW9662 against untreated control

PROVIDER: PRJEB56192 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR10228888.fastq.gz	Fastqsanger.gz
	ERR10228889.fastq.gz	Fastqsanger.gz
	ERR10228890.fastq.gz	Fastqsanger.gz
	ERR10228891.fastq.gz	Fastqsanger.gz
	ERR10228892.fastq.gz	Fastqsanger.gz

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Transcriptional profiling of human Th9 T cell clones treated with GW9662 against untreated control

Project description:To investigate the role of PPAR-γ in human TH cells, transcriptional response of activated “TH9” clones to treatment with GW9662, a potent PPAR-γ antagonist was assessed. “TH9” cell clones were incubated with GW9662 for 48h and activated with αCD3/2/28 for 12 h. Transcriptomic profiling was performed by bulk RNA-seq. To generate TH9 clones, Peripheral Blood Mononuclear Cells (PBMC) were isolated by Ficoll-Plaque Plus (GE Healthcare, UK) density gradient centrifugation. Human CD4+ T cells were isolated from PBMC by EasySep positive selection kit (Stemcell Technologies) according to manufacturer’s instruction. Positively selected CD4+ T cells were washed with PBS and stained for subsequent TH cell subset sorting. Effector memory “TH9” cells (CXCR3-CCR8+CCR6-CCR4+) were sorted to over 90% purity according to their expression of chemokine receptors from CD45RA-CD25-CD8-CD3+ cells. Single cell “TH9” clones were directly sorted into 96well plate according to their expression of chemokine receptors (see above). Single cell clones were expanded and maintained by periodic restimulation with PHA (1 µg/ml, Sigma-Chemicals) and irradiated allogenic feeder cells (5x104/well) in culture medium.

2022-10-31 | E-MTAB-12197 | biostudies-arrayexpress

Transcriptional profiling of human in vitro primed Th9 T cells (day 20) treated with GW9662 against untreated control

Project description:To investigate the role of PPAR-γ in human TH cells, transcriptional response in vitro primed Th9 cells to treatment with GW9662, a potent PPAR-γ antagonist was assessed. In vitro primed Th9 cells (day 20) were incubated with GW9662 for 48h. Transcriptomic profiling was performed by bulk RNA-seq. To generate in vitro primed Th9 cells, human naive T cells were isolated from PBMCs using the EasySep™ Human naive CD4+ T Cell Isolation Kit (Stemcell Technologies) as per the manufacturer’s instructions. Naive T cells were stimulated with αCD3/CD2/CD28 beads (T cell/bead = 2:1, Miltenyi) and primed into effector CD4+ Th9 cells with IL-4 (50 ng/ml) and TGF-β (5 ng/ml) (R&D Systems). From cell culture initiation to analysis, the culture medium was supplemented with the indicated cytokines every other day. Cells were harvested for RNA sequencing (RNA-seq) after 20 days.

2024-11-29 | E-MTAB-14444 | biostudies-arrayexpress

Transcriptional profiling of human in vitro primed Th9 T cells (day 20) treated with GW9662 against untreated control

Project description:Transcriptional profiling of human in vitro primed Th9 T cells (day 20) treated with GW9662 against untreated control

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Transcriptional profiling of Th1, Th2, Th9, and Th17 T cell clones

Project description:We next sought to identify the transcriptional program that differentiates IL-9+Th2 cells from “conventional” Th2 cells. To this end, we selected representative Th1, Th17, Th2, and IL-9+Th2 clones (Figure 5A) and determined their transcriptome in the resting state and at different time points after activation using RNAseq. Peripheral Blood Mononuclear Cells (PBMC) were isolated by Ficoll-Plaque Plus (GE Healthcare, UK) density gradient centrifugation. Human CD4+ T cells were isolated from PBMC by EasySep positive selection kit (Stemcell Technologies) according to manufacturer’s instruction. Positively selected CD4+ T cells were washed with PBS and stained for subsequent Th cell subset sorting. Memory Th cell subsets were sorted to over 90% purity according to their expression of chemokine receptors from CD45RA-CD25-CD8-CD3+ cells: Th1(CXCR3+CCR8-CCR6-CCR4-), Th2 (CXCR3-CCR8-CCR6-CCR4+), Th17 (CXCR3-CCR8-CCR6+CCR4+), Th9 (CXCR3-CCR8+CCR6-CCR4+). Single cell Th subset clones were directly sorted into 96well plate according to their expression of chemokine receptors (see above). Single cell clones were expanded and maintained by periodic restimulation with PHA (phytohaemaglutinine, 1 µg/ml, Sigma-Chemicals) and irradiated allogenic feeder cells (5x104/well) in culture medium. T cells were polyclonally activated using beads coated with antibodies against CD3, CD2, and CD28 (T cell/bead = 2:1, human T cell activation/expansion Kit, Miltenyi). Cell cultures were sampled before activation (time 0h) and 2, 4, 6, 9, 12, 24, and 48 hours after activation.

2017-11-10 | E-MTAB-5739 | biostudies-arrayexpress

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Project description:Microarray analyses were performed to compare gene expression in cultured mouse Th9, Th2 and Treg cells and resting versus activated Th9 cells. Three replicates were analyzed for each culture condition; Th9 unstim, Th2 unstim, Treg unstim, Th9 stim

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Transcriptional profiling of Th1, Th2, Th9, and Th17 T cell clones

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The transcription factor BATF3 regulates the differentiation of TH9 cells and TH9-driven mucosal pathologies

Project description:T helper 9 (TH9) cells are important for the development of inflammatory and allergic diseases. The TH9 transcriptional network converge signals from cytokines and antigen presentation but is incompletely understood. Here, we identified TL1A, a member of the TNF superfamily, as strong inducer of mouse and human TH9 differentiation. Mechanistically, TL1A induced the expression of the transcription factors BATF and BATF3 and facilitated their binding to the Il9 promoter leading to enhanced secretion of IL-9. BATF- and BATF3-deficiencies impaired IL-9 secretion under TH9 and TH9-TL1A polarizing conditions. In vivo, using a T cell transfer model we demonstrated that TL1A promoted IL-9-dependent, TH9 cell-induced intestinal and lung inflammation. Neutralizing IL-9 antibodies attenuated TL1A-driven mucosal inflammation. Batf3-/- TH9-TL1A cells induced reduced inflammation and cytokine expression in vivo compared to WT cells. Our results demonstrate that TL1A promotes TH9 cell differentiation and function and define a role of BATF3 in TH9-induced mucosal inflammation.

2019-01-08 | GSE106926 | GEO

Defining a Th9 transcriptome

Project description:Microarray analyses were performed to compare gene expression in cultured mouse Th9, Th2 and Treg cells and resting versus activated Th9 cells.

2014-03-26 | GSE44937 | GEO

LPS, Telmisartan and GW9662 co-treatment of microglial BV2 cells

Project description:LPS, Telmisartan and GW9662 co-treatment of microglial BV2 cells.

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Expression data from GW9662 treated FVB wild-type mouse mammary tumors

Project description:Microarrays were used to analyse global gene expression changes in tumors from wild-type mice treated with GW9662 RNA was isolated (RNeasy Mini Kit, Qiagen) from progestin/DMBA induced mammary tumors from wild-type mice treated with GW9662

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