E-GEOD-2405 - Transcription profiling time series of the human polymorphonuclear leukocyte response to exposure to Anaplasma phagocytophilum or Staphylococcus aureus

Status
Submitted on 14 March 2005, released on 25 August 2007, last updated on 2 May 2014
Organism
Homo sapiens
Samples (67)
Array (1)
Protocols (1)
Description
Polymorphonuclear leukocytes (PMNs) were obtained from healthy individuals in accordance with protocols approved by the Institutional Review Board for Human Subjects at the University of Minnesota and the National Institute of Allergy and Infectious Diseases. PMNs (107) were combined on ice with live S. aureus (108) or with live or heat-killed A. phagocytophilum (bacteria isolated from 5x106 infected HL60 cells for a ratio of 1 infected HL60 cell: 2 PMNs, ~ 5-20 A. phagocytophilum: PMN) in wells of a 12-well tissue culture plate (pre-coated with 20% autologous normal human serum). Unstimulated control assays received either buffer (for S. aureus comparisons) or clarified HL60 lysate (for A. phagocytophilum comparisons). Plates were centrifuged at 350 x g for 8 min at 4oC to synchronize phagocytosis and incubated at 37 deg. C in a CO2 incubator for the indicated times. At the indicated times, tissue culture medium was aspirated from the plate and PMNs were lysed directly with RLT buffer (Qiagen, Valencia, CA). Purification of PMN RNA and subsequent preparation of labeled cRNA target was performed as described in Methods. Labeling of samples, hybridization of cRNA with HU133A oligonucleotide arrays (Affymetrix, Santa Clara, CA), and scanning were performed according to standard Affymetrix protocols ( http://www.affymetrix.com/pdf/expression_manual.pdf ). Experiments were performed in triplicate, using PMNs from three healthy individuals for each treatment.
Experiment types
transcription profiling by array, co-expression, disease state, time series
Contact
Citation
Insights into pathogen immune evasion mechanisms: Anaplasma phagocytophilum fails to induce an apoptosis differentiation program in human neutrophils. Dori L Borjesson, Scott D Kobayashi, Adeline R Whitney, Jovanka M Voyich, Cynthia M Argue, Frank R Deleo. J Immunol 174(10):6364-72 (2005)
MIAME
PlatformsProtocolsFactorsProcessedRaw
Files
Investigation descriptionE-GEOD-2405.idf.txt
Sample and data relationshipE-GEOD-2405.sdrf.txt
Processed data (1)E-GEOD-2405.processed.1.zip
Array designA-AFFY-33.adf.txt
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