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E-GEOD-62373 - Age-associated Changes in Basal NF-κB Function in Human CD4+ T Lymphocytes via Dysregulation of PI3 Kinase (dataset 3)

Released on 8 January 2015, last updated on 9 January 2015
Homo sapiens
Samples (45)
Array (1)
Protocols (7)
Immune impairment and high circulating level of pro-inflammatory cytokines are landmarks of human aging. However, the molecular basis of immune dysregulation and the source of inflammatory markers remain unclear. Here we demonstrate that in the absence of overt cell stimulation, gene expression mediated by the transcription factor NF-κB is higher in purified and rested human CD4+ T lymphocytes from older compared to younger individuals. This increase of NF-κB -associated transcription includes transcripts for pro-inflammatory cytokines such as IL-1 and chemokines such as CCL2 and CXCL10. We demonstrate that NF-κB up-regulation is cell-intrinsic and mediated in part by phosphatidylinositol 3-kinase (PI3K) activity induced in response to metabolic activity, which can be moderated by rapamycin treatment. Our observations provide direct evidence that dysregulated basal NF-κB activity may contribute to the mild pro-inflammatory state of aging. Peripheral blood mononuclear cells (PBMCs) were isolated from heparinized blood using Ficoll-Paque Plus density gradient centrifugation. CD4+ T cells were obtained by positive selection using anti-human CD4 microbeads. After incubation for 4hr at either 4 or 37 Degrees C, cells were harvested and collected by centrifugation. Total cellular RNA was prepared using Qiagen RNeasy protocols. Labeling and amplification were done with the standard Illumina protocol and the biotin-labeled cRNA was hybridized to Illumina's HumanHT-12 V3.0 Expression BeadChip. The arrays were washed, blocked and the biotin-labeled cRNA was detected by staining with streptavidin-Cy3.
Experiment type
transcription profiling by array 
Kevin G Becker <>, Arsun Bektas, Elin Lehrmann, Karen Madara, Luigi Ferrucci, Ranjan Sen, William H Wood III, Yongqing Zhang
Investigation descriptionE-GEOD-62373.idf.txt
Sample and data relationshipE-GEOD-62373.sdrf.txt
Processed data (1)
Additional data (1)
Array designA-MEXP-1171.adf.txt