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Series GSE2405 Query DataSets for GSE2405
Status Public on May 15, 2005
Title NIH/NIAID Neutrophil Response to A. phagocytophilum
Organism Homo sapiens
Experiment type Expression profiling by array
Summary 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 ( ).

Experiments were performed in triplicate, using PMNs from three healthy individuals for each treatment.
Keywords: ordered
Web link
Contributor(s) Borjesson DL, Whitney AR, Kobayashi SD, Voyich JM, Argue CM, DeLeo FR, Argue CL
Citation(s) 15879137
Submission date Mar 14, 2005
Last update date Aug 10, 2018
Contact name Frank R. DeLeo
Phone 406-363-9448
Fax 406-363-9394
Organization name National Institute of Allergy and Infectious Diseases
Department Laboratory of Human Bacterial Pathogenesis
Lab Rocky Mountain Laboratories
Street address 903 South 4th Street
City Hamilton
State/province MT
ZIP/Postal code 59840
Country USA
Platforms (1)
GPL96 [HG-U133A] Affymetrix Human Genome U133A Array
Samples (67)
GSM45210 treatment=AP time=0 (hrs) AP1-0hr
GSM45211 treatment=AP time=1.5 (hrs) AP1-1.5hr
GSM45212 treatment=AP time=12 (hrs) AP1-12hr
BioProject PRJNA91613

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