Project description:Here we analyzed the transcriptional response to IL2 in NK cells in vitro. Splenic NK cells were isolated by negative selection from 6wk old male C57BL/6 mice. These were directly cultured with IL2 (250U/ml) for 8hrs in vitro. Cells were then transferred directly into TriZol. RNA was prepared in Trizol for gene expression profiling by Affymetrix Mouse Gene 1.0 ST Arrays. The samples in this series are re-analyzed samples from GSE84562.
Project description:IL2 signals are transmitted through JAK1 and JAK3, but the transcriptomic consequences of each to the overall response is unclear. Here we analyzed the relative contribution of JAK1 and JAK3 to the NK cell IL2 response in vitro using titrated doses of isoform specific JAK inhibitors. Blockade of JAK1 and JAK3 have unequal effects on IL2-induced transcripts at pharmacologically relevant doses. Splenic NK cells were isolated by negative selection from 6wk old male C57BL/6 mice. These were cultured with IL2 (250U/ml) in the presence or absence of JAK1/3 inhibitors for 8hrs in vitro. Cells were then transferred directly into TriZol. RNA was prepared in Trizol for gene expression profiling by Affymetrix Mouse Gene 1.0 ST Arrays.
Project description:IL2 signals are transmitted through JAK1 and JAK3, but the transcriptomic consequences of each to the overall response is unclear. Here we analyzed the relative contribution of JAK1 and JAK3 to the NK cell IL2 response in vitro using titrated doses of isoform specific JAK inhibitors. Blockade of JAK1 and JAK3 have unequal effects on IL2-induced transcripts at pharmacologically relevant doses.
Project description:Transcriptional profiling of NKAES-derived NK cells after 7 days of culture compared to primary human NK cells and NK cells stimulated by low or high dose IL2 after 7 days of culture. Four-condition experiment, primary NK cells vs. NKAES-derived NK cells after 7 days of culture vs. NK cells stimulated by low/high dose IL2 after 7 days of culture. Biological replicates: 5 control, 5 NKAES-derived NK cells, 3 NK cells stimulated by low dose IL2, 3 NK cells stimulated by high dose IL2 independently grown and harvested. One replicate per array.
Project description:Transcriptional profiling of NKAES-derived NK cells after 7 days of culture compared to primary human NK cells and NK cells stimulated by low or high dose IL2 after 7 days of culture.
Project description:The aim of the study was to investigate the activation of human NK cells by IL2 through analyzing the global gene expression at different time points (0, 2, 8 and 24 hours) after culture with the cytokine IL2 at 100 IU/ml. NK cells with the CD56+/CD16+ and CD3- phenotype were negatively selected by immunomagnetic beads and re-examined by flow-cytometry to ensure greater than 90% purity . Keywords: resting and IL2 activated NK cells(time series)
Project description:The aim of the study was to investigate the activation of human NK cells by IL2 through analyzing the global gene expression at different time points (0, 2, 8 and 24 hours) after culture with the cytokine IL2 at 100 IU/ml. NK cells with the CD56+/CD16+ and CD3- phenotype were negatively selected by immunomagnetic beads and re-examined by flow-cytometry to ensure greater than 90% purity . Experiment Overall Design: RNA was extracted and pooled from 4 different donors, amplified and labled according to the manufacturerâs instruction (GeneChipU133plus2® , Affymetrix Inc, CA).
Project description:Genome-wide transcriptional profiling was used to identify gene expression signatures and pathways in resting and IL2 activated NK cell isolated from peripheral blood of healthy donors Keywords: Time series_NK cells gene profile