Project description:Mouse spleen CD4+CD44LowCD62LHigh cells were stimulated with 2ug/ml anti-CD3(eBioscience), 2ug/ml anti-CD28 (eBioscience) in the presence (Th17 condition) or absence (Th0 condition) of hTGFb-1 (1ng/ml eBioscience), mouse IL-6(10ng/ml eBioscience). Cells were collected after 24 hourÕs culture using RNAeasy columns (Qiagen) for total RNA isolation. Isolated total RNA from Th0 and Th17 conditioned cells were submitted for the microarray gene transcription comparison analysis using Affymetrix Mouse 430A 2.0 array chips.
Project description:Stem cell memory T cells can be defined as the memory T cell subsets with naïve T cell phenotypes and stem cell-like properties. We find that Notch signaling converts activated T cells into stem cell memory T (TSCM) cells by OP9-DL1 feeder cell coculture. Based on gene expression profile analysis, we provide evidence that these cells represent distinct characters with central memory T (TCM) cells and naïve T cells with effector molecules and transcription factors. TSCM cells induced by Notch cluster closer to TCM cells than naïve T cells, and display similar phenotypes to TCM phenotype in terms of effector molecules and transcription factors, but the gene expressions of TSCM cells are entirely lower than that of TCM cells.
Project description:We compared CD8 TCM cells specific for a CMV epitope (pp65495-503) and an influenza A virus (IAV) epitope (M158-66) of the same healthy adults and identified genes whose expression are altered in CMV-specific compared to IAV-specific TCM cells.
Project description:Transcriptional profiling of CD8 Tcm cells comparing untreated control cells to 16 hour norepinephrine treated cells. The goal was to examine the impact of norepinephrine treatment on CD8 Tcm cell global gene expression compared to untreated cells, and also to examine changes that occurred in resting cells and activated cells 24 and 72 hours after stimulation with CD3/28 beads. Two-conditional experiment: Control vs. NE treated CD8 Tcm cells. Biological replicates: 3 replicates for 0 hour and 24 hours, and 2 replicates for 72 hours. Each sample consisted of pooled RNA from 3 different donors.
Project description:The design of this experiment was based on the assumption that transcript levels will change linearly while going from 100 percent of one tissue to 100 percent of the other. This assumed linear data set could be used to evaluate various issues related to low-level microarray data analysis. Hybridization cocktails from two mouse tissues, kidney and spleen, were prepared and mixed in a range of ratios and applied to 4-5 replicate GLYCOv1 chips for analysis. The mixture ratios were as follows: 100 percent kidney, 75 percent Kidney/25 percent spleen, 50 percent kidney/50 percent spleen, 25 percent kidney/75 percent spleen, and 100 percent spleen.
