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RNA squencing of human NK cells activated with IL-15, a combination of IL-12, IL-15, and IL-18, or with a trimeric fusion protein 18/12/TxM

ABSTRACT: RNA squencing of human NK cells activated with IL-15, a combination of IL-12, IL-15, and IL-18, or with a trimeric fusion protein 18/12/TxM

PROVIDER: PRJNA699884 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13642627_1.fastq.gz	Fastqsanger.gz
	SRR13642627_2.fastq.gz	Fastqsanger.gz
	SRR13642628_1.fastq.gz	Fastqsanger.gz
	SRR13642628_2.fastq.gz	Fastqsanger.gz
	SRR13642629_1.fastq.gz	Fastqsanger.gz

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RNA squencing of human NK cells activated with IL-15, a combination of IL-12, IL-15, and IL-18, or with a trimeric fusion protein 18/12/TxM

Project description:Gene expression analysis of human NK cells at baseline, Day 1, and Day 6 after activation with IL-15, IL-12/15/18, or the 18/12/TxM molecule

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RNA squencing of human NK cells activated with IL-15 or IL-12, IL-15, and IL-18

Project description:RNA squencing of human NK cells activated with IL-15 or IL-12, IL-15, and IL-18

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RNA squencing of human NK cells activated with IL-15 or IL-12, IL-15, and IL-18

Project description:NK cells are an emerging cancer cellular therapy and potent mediators of anti-tumor immunity. Cytokine-induced memory-like (ML) NK cellular therapy is safe and induces remissions in acute myeloid leukemia (AML) patients. However, the dynamic molecular changes that occur after memory-like differentiation in vitro are unclear. Here, control or ML NK cells purified from normal donor PBMC were generated in vitro. Briefly, RosetteSep-purified NK cells were incubated in IL-12, IL-15, and IL-18, or low-dose IL-15 as a control for 16-18 hours. Control or cytokine-activated NK cells were washed three times and cultured for 6 days in low-dose IL-15, which is required for NK cell survival. After 6 days, RNA was isolated from control and memory-like (ML) NK cells (IL12/15/18 activation) and RNA-sequencing performed. Because the transcription factor GATA-3 was increased specifically in ML NK cells, we hypothesized ML NK cells would exhibit a GATA-3 gene signature compared to control NK cells. Indeed, using GSEA, a significant gene signature was associated with ML NK cell differentiation. These data support the role for GATA-3 in regulating the ML NK cell molecular program.

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Transcriptional changes in IL-2, IL-12 and IL-18 stimulated human natural killer cells

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Effect of IL-12+IL-18 or anti-NK1.1 stimulation on splenic NK cells [mNK_Activation_RNA-seq]

Project description:We stimulated freshly enriched splenic NK cells with either IL-12+IL-18 (1ng/ml each) or plate-bound anti-NK1.1. NKs from several C57Bl/6 mice were pooled. We then performed gene expression profiling analysis using data obtained from RNA-seq at two time points: 3 or 6 hours.

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Glucocorticoids and the cytokines IL-12, IL-15 and IL-18 present in the tumor microenvironment induce PD-1 expression on human Natural Killer cells

Project description:Background: PD-1 immune checkpoint blockade has provided significant clinical efficacy across various types of cancer by unleashing both T- and NK cell-mediated anti-tumor responses. However, resistance to immunotherapy occurs for many patients, rendering the identification of the mechanisms that control PD-1 expression extremely important to increase the response to the therapy Objective: To identify the stimuli and the molecular mechanisms that induce the de novo PD-1 expression on human NK cells in the tumor setting Methods: NK cells freshly isolated from peripheral blood of healthy donors were stimulated with different combinations of molecules, and PD-1 expression was studied at the mRNA and protein level. Moreover, ex vivo analysis of tumor microenvironment and NK cell phenotype was performed. Results: Glucocorticoids (GCs) are indispensable for PD-1 induction on human NK cells, in cooperation with a combination of cytokines that are abundant at the tumor site. Mechanistically, GCs together with IL-12, IL-15 and IL-18 not only upregulate PDCD1 transcription, but also activate a previously unrecognized transcriptional program leading to enhanced mRNA translation and resulting in an increased PD-1 protein amount in NK cells. Conclusion: Our results provide evidence of a novel immune suppressive mechanism of GCs involving the transcriptional and translational control of an important immune checkpoint

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Transcriptional changes in IL-2, IL-12 and IL-18 stimulated human natural killer cells

Project description:We used Affymetrix expression arrays to determine changes in gene expression associated with activation of human NK cells mediated through treatment with cytokines IL-2, IL-12 and IL-18 over a 24 hour period. Human natural killer cells were isolated via negative selection from PBMCs of healthy donors. Cells were found to be > 95% CD3-CD56+. RNA was harvested at time of isolation or after 24 hour stimulation from 8 x 10^6 cells per condition. For stimulations, cells were incubated at 37C (5% CO2) in RPMI, suuplemented with 10% Fetal Bovine Serum at 1.5 x 10^6 cells/ml. Cytokine stimualtions were conducted with IL-2 (100U/mL), IL-12 (10ng/mL) and IL-18 (100ng/mL) from 2 male donors (N = 4). Expression analysis was carried out to determine transcriptional changes associated with 24 hr stimulation relative to freshly isolated cells.

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Glucocorticoids and the cytokines IL-12, IL-15 and IL-18 present in the tumor microenvironment induce PD-1 expression on human Natural Killer cells

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Transcription profiling of mouse IL-12+IL-18 treated and Yersinia enterocolitica infected C57BL/6 NK cells

Project description:Dataset of IL-12+IL-18 trated and Yersinia enterocolitica infected C57BL/6 NK cells Experiment Overall Design: NK cells were isolated from mouse spleen, grown on IL-2 in vitro, stimlated wih Y. enterocolitica WA(pYV) or left uninfected. Experiment Overall Design: 3 conditions, 3 biological replicates each

2009-01-13 | E-GEOD-9735 | biostudies-arrayexpress

Dissection of IL-2 and IL-15 signaling pathways

Project description:The aim of this study was to compare the signaling cascades initiated by the two closely related cytokines IL-2 and IL-15. Considering the relevance of protein tyrosine phosphorylation in signal transduction, in order to quantify changes in protein phosphorylation in response to IL-2 and IL-15, we combined triple SILAClabeling of leukemic T-cells with phosphotyrosine (pY)-specific antibody-based protein enrichment followed by mass spectrometry (MS) analysis. Following the strategy described above, we managed to decipher in detail the complex signaling networks triggered by IL-2 and IL-15. Interestingly, a large number of components of the three main signaling pathways known to be initiated in response to the interleukins (JAK/STAT; RAS/MAPK; PI3K/AKT) were identified.

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