|Application:||Gene expression microarray analysis|
|Number of samples:||80|
|Release date:||Jan 8 2010|
|Last update date:||Jul 10 2014|
|Dataset link||Genome Wide Analysis of Immune Activation in Human T and B Cells Reveals Distinct Classes of Alternatively Spliced Genes|
CD2+ T-lymphocytes and CD19+ B-lymphocytes were purified from peripheral blood mononuclear cells (PBMC) of 10 normal human donors. Freshly isolated, resting CD2+ T-cells were resuspended in RPMI-1640 complete media and activated with CD3/CD28 Dynal beads. Cultures were sampled at 24, 48 and 72hrs. A subset of isolated, resting cells immediately stabilized by RNALater solution were used as the baseline comparison (Time 0). Freshly isolated CD19+ magnetic-bead purified resting B cells were resuspended in RPMI-1640 complete media to which anti-CD40 antibody was added, followed by cross-linking with goat-anti-mouse anti-IgG1. The cells were then cultured with rIL2 and rIL10 (100 ng/ml each) to simulate the T-cell dependent B cell activation of an alloimmune response. RNA was extracted at time 0, 24, 48 and 72 hours post activation using the mirVana miRNA Isolation Kit. The GeneChip® The RNA was converted into labeled cDNA using the GeneChip® WT Sense Target Labeling kit (Affymetrix). Labeled cDNA was hybridized to Affymetrix Human Exon 1.0 ST arrays. Data for mRNA transcript profiles were generated in the form of CEL files using standard protocols (http://www.affymetrix.com/).
Sunil Mathan Kurian
Citations per year