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:We used microarrays to detail the global programme of gene expression by circulating TCRVgamma9+ gamma delta T cells isolated from healthy individuals,tested either as resting cells or cells activated by phosphoantigen BrHPP and IL-2at an early(+6hrs) and a late (+7days) timepoint. We find that with more M-bM-^@M-^\NK cellM-bM-^@M-^] genes than alphabeta T cells and more M-bM-^@M-^\T cellM-bM-^@M-^] genes than NK cells, the circulating TCRVgamma9+ gamma delta T cells cells have a hybrid transcriptome. The gene signature of the activated cells recapitulates their physiological functions: Th1 cytokine, chemokine and cytotoxic activities at first and mitotic activity at later time points. The gene expression pattern of activated normal gamma delta T cells is nevertheless clearly distinctive from that of NK/T and peripheral T cell lymphomas of the gamma delta subtype. Human TCRVg9positive gamma delta T cells were isolated from PBMC by cell sorting (>98% purity) and activated for RNA extraction and hybridization on Affymetrix microarrays. Samples comprise cells before activation (control time 0), early after activation with BrHPP/IL2 (+6 hours) and at a later timepoint of the activated in vitro culture with BrHPP/IL2 (day 7).
Project description:Pancreatic ductal adenocarcinoma (PDAC) remains a particularly aggressive disease with few effective treatments. The PDAC tumor immune microenvironment (TIME) has been characterized as immune suppressed. Oncolytic viruses can increase tumor antigenicity via immunogenic cell death (ICD). In this study, tumor-targeting and cytokine-armed vaccinia viruses (vvDD, vvDD-IL2, vvDD-IL15) were used to infect carcinoma cell lines as well as patient-derived primary PDAC cells. In co-culture experiments we tested the cytotoxic response and the activation of human natural killer-(NK-)cells during the oncolytic process.
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:Low-Grade Glioma RCAs mice were treated with one dose of Vehicle (PBS) or GEMys-IL2 We analized the tumor-infiltrating immune cells at day 3 post-treatment with GEMys-IL2, or Vehicle.
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: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
Project description:CD25 (IL2RA) is a subunit of IL2 receptor complex, which determines the sensitivity of the receptor to IL2. Here, we investigated whether PRDM1, a transcriptional repressor implicated as a tumor suppressor in NK cell malignancies, directly represses CD25 (IL2RA). Using ChIP-seq, we identified a direct binding site of PRDM1 within 1st intron of CD25 (IL2RA) in activated primary human NK cells. We used DNA microarrays on two PRDM1α transduced NK cell lines (i.e. NK92 and KHYG1) to address whether PRDM1 transcriptionally represses CD25.