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

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

PROVIDER: PRJEB79960 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR13656139.fastq.gz	Fastqsanger.gz
	ERR13656140.fastq.gz	Fastqsanger.gz
	ERR13656141.fastq.gz	Fastqsanger.gz
	ERR13656142.fastq.gz	Fastqsanger.gz
	ERR13656143.fastq.gz	Fastqsanger.gz

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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 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.

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

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

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Transcriptional profiling of human in vitro primed Th1, Th2, Th9 and Treg T cells

Project description:To test whether human in vitro primed Th9 cells recapitulate the core pathogenic Th2 cell phenotype, we differentiated naïve T cells into Th1 (IL-12), Th2 (IL-4), Th9 (IL-4+TGF-β), and iTreg (TGF-β). After 7 days transcriptomic profiling by bulk RNA-seq was performed.

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

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