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Series GSE2396 Query DataSets for GSE2396
Status Public on Jul 20, 2005
Title PBS vs. LPS stimulation of gd T cells
Organism Homo sapiens
Experiment type Expression profiling by array
Summary gd T cells recognize unprocessed or non-peptide antigens, respond rapidly to infection, and localize to mucosal surfaces. We have hypothesized that the innate functions of gd T cells may be more similar to those of cells of the myeloid lineage than to other T cells. To begin to test this assumption, we have analyzed the direct response of cultured human and peripheral blood bovine gd T cells to pathogen associated molecular patterns (PAMPs) in the absence of APCs using microarray, real time RT-PCR, proteome array, and chemotaxis assays. Our results indicate that purified gd T cells respond directly to PAMPs by increasing expression of chemokine and activation related genes. The response was distinct from that to known gd T cell antigens and different from the response of myeloid cells to PAMPs. In addition, we have analyzed the expression of a variety of PAMP receptors in gd T cells. Freshly purified bovine gd T cells responded more robustly to PAMPs than did cultured human cells and expressed measurable mRNA encoding a variety of PAMP receptors. Our results suggest that rapid response to PAMPs through the expression of PAMP receptors may be another innate role of gd T cells.
Keywords: parallel sample
 
Overall design Sorted cells from 4 human gd T cell cultures were treated with either PBS or phLPS for 4 hours, RNA was extracted with TRIzol® Reagent (Invitrogen Life Technologies) according to the manufacturer’s instructions, pooled and used to probe Affymetrix Genechip® Human Genome U133A 2.0 Arrays (Cat. # 900471 Affymetrix, Santa Clara, CA) that represent 14,500 human genes. cDNA amplification and synthesis of biotin-labeled cRNA was performed with the Alternative One-cycle target labeling protocol with 10 micrograms total RNA as described in the GeneChip® Expression Analysis Technical Manual (March 2004). Hybridization was performed with 10 micrograms biotin labeled cRNA. Washing and staining was performed in the GeneChip® Fluidics Station 450 using the Midi_euk2v3 protocol. Chip scans were performed on the Affymetrix GeneChip® Scanner 3000. GeneChip® Operating Software (GCOS v.1.1, Affymetrix) was used for data collection.
 
Contributor(s) Hedges JF, Lubick KL, Jutila MA
Citation(s) 15879098
Submission date Mar 10, 2005
Last update date Dec 06, 2018
Contact name Jodi Fern Hedges
E-mail(s) [email protected]
Phone 406-994-6384
Organization name Montana State University
Department Veterinary Molecular Biology
Lab Jutila
Street address 960 Technology Blvd.
City Bozeman
State/province MT
ZIP/Postal code 59718
Country USA
 
Platforms (1)
GPL571 [HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array
Samples (2)
GSM45116 human gd PBS
GSM45117 human gd LPS
Relations
BioProject PRJNA91597

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE2396_RAW.tar 6.0 Mb (http)(custom) TAR (of CEL)

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