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Status |
Public on Jul 20, 2005 |
Title |
PBS vs. LPS stimulation of gd T cells |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by array
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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
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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.
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Contributor(s) |
Hedges JF, Lubick KL, Jutila MA |
Citation(s) |
15879098 |
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Submission date |
Mar 10, 2005 |
Last update date |
Dec 06, 2018 |
Contact name |
Jodi Fern Hedges |
E-mail(s) |
[email protected]
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Phone |
406-994-6384
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Organization name |
Montana State University
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Department |
Veterinary Molecular Biology
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Lab |
Jutila
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Street address |
960 Technology Blvd.
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City |
Bozeman |
State/province |
MT |
ZIP/Postal code |
59718 |
Country |
USA |
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Platforms (1) |
GPL571 |
[HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array |
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Samples (2) |
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Relations |
BioProject |
PRJNA91597 |
Supplementary file |
Size |
Download |
File type/resource |
GSE2396_RAW.tar |
6.0 Mb |
(http)(custom) |
TAR (of CEL) |
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