Expression data measured by custom Nanostring gene set of CD4+ T cells from healthy individuals stimulated with anti-CD3/CD28 with or without IFNb or Th17 polarizing cytokines
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ABSTRACT: Variation in individuals' adaptive immune response is believed to influence susceptibility to complex diseases in humans. The genetic basis of such variation is poorly understood. We measured gene expression from resting and activated CD4+ T cells derived from the peripheral blood of 348 healthy individuals. We activated the primary T cells with anti-CD3/CD28 beads. We collected peripheral blood from each human donor. We isolated peripheral blood mononuclear cells by Ficoll, and negatively selected for CD4+ T cells using RosettaSep. We isolated peripheral blood mononuclear cells by Ficoll, and negatively selected for CD4+ T cells using RosettaSep. We then either left cells unstimulated or stimulated them with beads conjugated with anti-CD3 and anti-CD28 either without additional cytokines, or with IFNb, or with Th17 cocktail. Cells were harvest at 0hr, 4hr (anti-CD3/CD28 +/- IFNb) or 48hr (anti-CD3/CD28 +/- Th17), lysed and RNA isolated to be profiled on Nanostring.
Project description:Variation in individuals' adaptive immune response is believed to influence susceptibility to complex diseases in humans. The genetic basis of such variation is poorly understood. We measured gene expression from resting and activated CD4+ T cells derived from the peripheral blood of healthy individuals. We activated the primary T cells with anti-CD3/CD28 beads alone or with IFNb or Th17 polarizing cytokines. We collected peripheral blood from each human donor. We isolated peripheral blood mononuclear cells by Ficoll, and negatively selected for CD4+ T cells using RosettaSep. We then either left cells unstimulated or stimulated them with beads conjugated with anti-CD3 and anti-CD28 either without additional cytokines, or with IFNb, or with Th17 cocktail. Cells were harvest at up to 8 time points (0hr, 45min, 2hr, 4hr, 10hr, 24hr, 48hr and 72hr), lysed and RNA isolated to be profiled on microarray.
Project description:Variation in individuals' adaptive immune response is believed to influence susceptibility to complex diseases in humans. The genetic basis of such variation is poorly understood. We measured gene expression from resting and activated CD4+ T cells derived from the peripheral blood of healthy individuals. We activated the primary T cells with anti-CD3/CD28 beads alone or with IFNb or Th17 polarizing cytokines.
Project description:Variation in individuals' responses to environmental factors is believed to influence susceptibility to complex diseases in humans. The genetic basis of such variation is poorly understood. We measured gene expression from resting and stimulated dendritic cells (DCs) derived from the peripheral blood of healthy individuals. We stimulated the primary DCs with E. coli lipopolysaccharide (LPS) or influenza virus. Using serial replicate samples, we selected genes that showed evidence of reproducibility within the serial replicates. We collected peripheral blood from each human donor. We isolated peripheral blood mononuclear cells by Ficoll, and magnetically sorted them for CD14+CD16- monocytes. We then differentiated the monocytes into monocyte-derived dendritic cells (MoDCs) by culturing the cells for 7 days with GM-CSF and IL-4. We stimulated the cells with E. coli lipopolysaccharide (LPS) for 5 hr or influenza (PR8 dNS1) for 10 hr. Finally, we lysed the cells and isolated total RNA for microarray.
Project description:This SubSeries in the ImmVar project investigates the response of selected genes in T cells from healthy human individuals to ascertain the impact of genetic or non-genetic variation on T cell activation parameters. We measured gene expression from resting and activated CD4+ T cells derived from the peripheral blood of healthy individuals. We activated the primary T cells with anti-CD3/CD28 beads alone or with IFNb or Th17 polarizing cytokines.
Project description:To indentify the effect of JPH203 on the gene expression in activated T cells, microarray analysis was performed using activated human T cells derived from peripheral blood. Human CD4+ T cells from peripheral blood in healthy volunteer were activated with anti-CD3 and anti-CD28 in the presence of 1uM JPH203 for 3days. RNA was extracted and gene expression was compared between JPH203-treated cells and control cells. Gene epression of human peripheral blood CD4+ T cells activated with anti-CD3 and anti-CD28 in the presence of 1uM JPH203 for 3days was measured.
Project description:Variation in individuals' adaptive immune response is believed to influence susceptibility to complex diseases in humans. The genetic basis of such variation is poorly understood. We measured gene expression from resting and activated CD4+ T cells derived from the peripheral blood of healthy individuals. We activated the primary T cells with anti-CD3/CD28 beads. We collected peripheral blood from each human donor. We isolated peripheral blood mononuclear cells by Ficoll, and negatively selected for CD4+ T cells using RosettaSep. We then either left cells unstimulated or stimulated them with beads conjugated with anti-CD3 and anti-CD28. Cells from 15 individuals were harvested at up to 3 time points (0hr, 4hr or 48hr), lysed and RNA isolated to be profiled on microarray.
Project description:To indentify the effect of JPH203 on the gene expression in activated T cells, microarray analysis was performed using activated human T cells derived from peripheral blood. Human CD4+ T cells from peripheral blood in healthy volunteer were activated with anti-CD3 and anti-CD28 in the presence of 1uM JPH203 for 3days. RNA was extracted and gene expression was compared between JPH203-treated cells and control cells.
Project description:To examine the effects of prostaglandin E2/EP4 signaling on bovine CD4+ T cells, CD4+ T cells were isolated from bovine peripheral blood mononuclear cells. Isolated CD4+ T cells were cultured with anti-bovine CD3 and anti-bovine CD28 monoclonal antibodies. Following 18 hours of incubation, the cultures were incubated with EP4 agonist or DMSO, a vehicle control for 4 hours. After incubation, the cells were collected and microarray analysis was performed.
Project description:Variation in individuals' adaptive immune response is believed to influence susceptibility to complex diseases in humans. The genetic basis of such variation is poorly understood. We measured gene expression from resting and activated CD4+ T cells derived from the peripheral blood of 348 healthy individuals. We activated the primary T cells with anti-CD3/CD28 beads.
Project description:microRNA transcriptional profiling of human naïve CD4 T cells comparing cells that saw no activation, only anti-CD3 or anti-CD3 and anti-CD28 to identify microRNAs specifically upregulated by CD28-mediated costimulation. Three conditions experiment. PBS, anti-CD3, anti-CD3 + anti-CD28 all compared to a reference sample. Biological triplicates, independently extracted and activated.