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Transcription profiling of human PMNs to compare global gene expression in human neutrophils during spontaneous apoptosis with that in cells cultured with human GM-CSF


ABSTRACT: PMNs were isolated from venous blood of healthy individuals in accordance with a protocol approved by the Institutional Review Board for Human Subjects, National Institute of Allergy and Infectious Diseases. Human PMNs (107) were cultured in RPMI/H -/+ 100 ng/ml GM-CSF at 37°C and 5% CO2 for up to 24 h as indicated. At the indicated time points, tissue culture medium was aspirated from each well and PMNs were lysed with RLT buffer (Qiagen). Purification of PMN RNA and subsequent preparation of labeled cRNA target was performed as described in Methods. Labeling of samples, GeneChip hybridization (Hu95Av2 oligonuceotide microarrays) and scanning were performed according to standard Affymetrix protocols (see http://www.affymetrix.com/support/technical/manual/expression_manual.affx). Experiments were performed in triplicate, using PMNs from three healthy individuals for each treatment. These analyses allowed comparison of global gene expression in human neutrophils during spontaneous apoptosis with that in cells cultured with human GM-CSF. Experiment Overall Design: 39 samples total: 9 time zero controls, -/+ GM-CSF at 5 time points in triplicate

ORGANISM(S): Homo sapiens

SUBMITTER: Addie Whitney 

PROVIDER: E-GEOD-2803 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Spontaneous neutrophil apoptosis and regulation of cell survival by granulocyte macrophage-colony stimulating factor.

Kobayashi Scott D SD   Voyich Jovanka M JM   Whitney Adeline R AR   DeLeo Frank R FR  

Journal of leukocyte biology 20051004 6


Polymorphonuclear leukocytes (PMNs or neutrophils) are the most prominent cellular component of the innate immune system in humans and produce an array of potent cytotoxic molecules. It is important that neutrophils undergo constitutive (spontaneous) apoptosis as a mechanism to facilitate normal cell turnover and immune system homeostasis. Conversely, several proinflammatory cytokines, including granulocyte macrophage-colony stimulating factor (GM-CSF), prolong neutrophil survival. The molecular  ...[more]

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