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: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.
Project description:In this study we have compared the proteomic profile of extracellular vesicles (EVs) prepared from primary, human NK cells or the human NK cell lines NK-92 and KHYG-1 cultured for 48hrs in serum-free conditions. EVs were harvested from cells either under resting conditions (culture in IL-15) or upon activation (combination of IL-12, IL-15, and IL-18). In addition, primary NK cells were activated in the presence of anti-CD16-coated beads, and EVs harvested after 48hrs. The aim was to compare their ability to target and kill a variety of tumor cell line-derived spheroids
Project description:Ex vivo activation and expansion of natural killer (NK) cells is a strategy to produce sufficient numbers of these effector cells for adoptive immunotherapy. Therefore we aimed at the development of an automated, clinical scale NK cell expansion process and compared NK cells after manual and automated expansion.We found only a small subset of 124 genes that significantly varied between both sample groups. These genes were associated with movement of leukocytes were identified including a group of genes for NK cell movement (CMKLR1, CX3CR1, S1PR5, GNLY and CXCR1) which were slightly higher expressed after automated compared to manual expansion. Nevertheless, the expansion profiles of NK cells after automated or manual expansion were rather similar in comparison to the remarkable difference between primary and expanded NK cells in general represented by the vast majority of regulated genes between the analyzed sample groups. Pathway analysis revealed that most of regulated genes upon expansion were associated with cell cycle regulation, DNA replication, DNA recombination and DNA repair, regulation of apoptosis and cell survival as well as cytokine signaling. Furthermore, the expression of many NK cell relevant markers was changed upon expansion with the strongest effects on up regulation of TRAIL and FASL, inhibitory TIGIT and chemokine receptors CCR2, CCR5 and CXCR6. In addition, granzyme M was down regulated but other important effector molecules like TNFa, perforin and granzymes A, B and K were up regulated. In summary we could show that manual and automatically expanded NK cells show a similar expansion profile while the differ significantly from primary NK cells. Up regulation of many NK cell relevant markers indicate for an enhanced NK cell functionaility after ex vivo activation and expansion. 24 samples were analyzed in total. 6 biological replicates represented by cells from different donors. 4 different samples per donor: freshly isolated NK cells (d0) and NK cells expanded for 14 days under 3 different conditions: in co-culture with EBV-LCL feeder cells and 500 U/mL IL2 either cultivated manually in T75 flasks (T) or automatically using the CliniMACS Prodigy system (P); NK cells cultured with 500 U/ml IL2 without feeder cells (I)