Project description:This SuperSeries is composed of the SubSeries listed below. 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 analyzed the response of human CD4+ T cells from a cohort of 348 healthy humans, representing three major ancestries, to in vitro stimulation, alone or in Th17 conditions, or with IFNb stimulation.
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:Little is known about the global transcriptional program underlying CD4+ T-cell activation. Using DNA microarrays and Q-RT-PCR, we examined the transcriptional profile of human CD4+ T-cell activation. The goal of this study was to identify genes involved in the various facets of human CD4+ T-cell activation. Experiment Overall Design: CD4+ T cells isolated from peripheral blood were cultured with CD3, CD28, with or without IL-2 to induce T-cell activation. At each timepoint, cells were harvested and frozen for RNA isolation. Three biological replicate experiments were analyzed and approximately one-half of the samples from each experiment were technically replicated. Hybridizations were performed in a reference design with all samples labeled with Cy3 and a reference thymus total RNA labeled with Cy5.
Project description:Little is known about the global transcriptional program underlying CD4+ T-cell activation. Using DNA microarrays and Q-RT-PCR, we examined the transcriptional profile of human CD4+ T-cell activation. The goal of this study was to identify genes involved in the various facets of human CD4+ T-cell activation. Keywords: time course
Project description:We analyzed the total proteome of CD4+ T cells isolated from WT mice, either non stimulated or at 5 different time points of stimulation with anti-CD3 and anti-CD4 antibodies (30s; 120s; 300s; 600s)
Project description:In people with HIV (PWH), the post antiretroviral therapy (ART) window is critical for immune restoration and HIV reservoir stabilization. We employ deep immune profiling, T cell receptor (TCR) sequencing and examine proliferation to assess how ART impacts T cell homeostasis. Hierarchies and frequencies of dominant CD4 TCR clonotypes (0.1-11% of all CD4+ T cells) remain stable post-ART, suggesting that clonal homeostasis can be independent of homeostatic processes regulating CD4+T cell absolute number, phenotypes and function. The slow restoration of host-immunity post-ART also has implications for the design of ART interruption studies.