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Effect of FcRγ on gene expression profile of naïve and IL-15 treated NK cells

ABSTRACT: Effect of FcRγ on gene expression profile of naïve and IL-15 treated NK cells

PROVIDER: PRJNA1212070 | ENA |

REPOSITORIES: ENA

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Effect of FcRγ on gene expression profile of naïve and IL-15 treated NK cells

Project description:Here, we aimed to study the NK cell molecular signatures under the control of FcRγ and how these signatures are regulated by IL-15 and the tumour microenvironment (TME). To investigate this, we obtained purified splenic NK cells from naïve wild-type (WT) and FcRγ-deficient (FcRγ–/–, KO) mice cultured in the presence or absence of IL-15 for three days. To obtain NK cells from the TME, NK cells were purified from tumour-bearing lungs of WT and KO (FcRγ–/–) mice, which were intravenously injected with B16F10 cells two weeks earlier. Naïve NK cells (without any IL-15 treatment) were used as the control group. Next, we performed bulk RNA sequencing (RNA-seq) on six groups of NK cells for gene expression profiling: naive WT, naive KO (FcRγ–/–), IL-15-treated WT, IL-15-treated KO (FcRγ–/–), tumour WT, and tumour KO (FcRγ–/–).

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Phosphoproteomics of Natural Killer (NK) Cells Stimulated with Fibrin and IL-15

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Runx3 function in splenic NK cells (IL-2 or IL-15)

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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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Runx3 function in splenic NK cells (IL-2 or IL-15)

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mRNA expression profiles of resting and IL-15 activated murine NK cells

Project description:Murine NK cells were compared at rest and following 24 hours of IL-15 stimulation for their mRNA expression profiles on the Affymetrix MOE430_2 microarray platform. Additional comparators included resting bulk splenocytes. Keywords: mRNA, mouse, NK cell, IL-15

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Project description:Splenic NK cells were enriched and cultured in either low dose (5-10 ng/ml) or high dose (100 ng/ml) IL-15. Naïve NKs are freshly enriched NKs, obtained the day of the anti-NK1.1 stimulation. Cells were either left unstimulated or were stimulated via plate-bound anti-NK1.1

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Runx3 function in splenic NK cells activated in vivo by IL-15.

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Runx3 function in splenic NK cells activated in vivo by IL-15.

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