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Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects.

Abstract：

:Administration of endotoxin (LPS) in humans results in profound physiological responses, including activation of peripheral blood mononuclear cells and the release of inflammatory factors. The time course of the response of selected inflammatory proteins was examined in healthy subjects (n = 6) administered a single intravenous dose of the purified derivative of endotoxin (3.0 ng/kg). Microarray analysis demonstrated changes in the expression of a number of genes, which were confirmed in separate in vitro endotoxin stimulation experiments. Subsequent TaqMan analysis of genes of interest indicated time-dependent changes in the expression of many of these genes. This included pre-B cell enhancing factor, which was identified on microarray analysis as being markedly upregulated following endotoxin stimulation. Protein expression of the genes examined by TaqMan analysis was measured and demonstrated the appearance of tumor necrosis factor (TNF)-alpha and sTNF-R proteins in the plasma beginning within 1 h after dosing, followed by other cytokines/ inflammatory markers (e.g., IL-1ra, G-CSF, IL-6, IL-8, and IL-10) and suppressors of cytokine signaling (SOCS-1 and SOCS-3). In general, cytokine protein expression correlated well with gene expression; however, the temporal profile of expression of some genes did not correlate well with the protein data. For many of these proteins, the lack of correlation was attributable to alternate tissue sources, which were demonstrated on TaqMan analysis. Principal component analysis indicated that cytokines could be grouped according to their temporal pattern of response, with most transcript levels returning to baseline 24 h following endotoxin administration. The combination of cDNA microarray and TaqMan analysis to identify and quantify changes in gene expression, along with the analysis of protein expression, can be useful in investigating inflammatory and other diseases.

journal_name

DNA Cell Biol

journal_title

DNA and cell biology

authors

Prabhakar U,Conway TM,Murdock P,Mooney JL,Clark S,Hedge P,Bond BC,Jazwinska EC,Barnes MR,Tobin F,Damian-Iordachi V,Greller L,Hurle M,Stubbs AP,Li Z,Valoret EI,Erickson-Miller C,Cass L,Levitt B,Davis HM,Jorkasky DK

doi

10.1089/dna.2005.24.410

keywords:

subject

Has Abstract

pub_date

2005-07-01 00:00:00

pages

410-31

issue

eissn

1044-5498

issn

1557-7430

journal_volume

pub_type

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Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects.