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Impact of anti-inflammatory agents on the gene expression profile of stimulated human neutrophils: unraveling endogenous resolution pathways.

St-Onge M, Dumas A, Michaud A, Laflamme C, Dussault AA, Pouliot M - PLoS ONE (2009)

Bottom Line: Genes encoding transcription factors, enzymes and regulatory proteins, as well as secreted cytokines/chemokines showed differential expression.We have identified a set of genes that may be part of important resolution pathways that interfere with cell activation.Identification of these pathways will improve understanding of the capacity of tissues to terminate inflammatory responses and contribute to the development of therapeutic strategies based on endogenous resolution.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy-Physiology, Faculty of Medicine, Centre de Recherche en Rhumatologie et Immunologie du CHUQ, Laval University, Quebec City, Quebec, Canada.

ABSTRACT
Adenosine, prostaglandin E(2), or increased intracellular cyclic AMP concentration each elicit potent anti-inflammatory events in human neutrophils by inhibiting functions such as phagocytosis, superoxide production, adhesion and cytokine release. However, the endogenous molecular pathways mediating these actions are poorly understood. In the present study, we examined their impact on the gene expression profile of stimulated neutrophils. Purified blood neutrophils from healthy donors were stimulated with a cocktail of inflammatory agonists in the presence of at least one of the following anti-inflammatory agents: adenosine A(2A) receptor agonist CGS 21680, prostaglandin E(2), cyclic-AMP-elevating compounds forskolin and RO 20-1724. Total RNA was analyzed using gene chips and real-time PCR. Genes encoding transcription factors, enzymes and regulatory proteins, as well as secreted cytokines/chemokines showed differential expression. We identified 15 genes for which the anti-inflammatory agents altered mRNA levels. The agents affected the expression profile in remarkably similar fashion, suggesting a central mechanism limiting cell activation. We have identified a set of genes that may be part of important resolution pathways that interfere with cell activation. Identification of these pathways will improve understanding of the capacity of tissues to terminate inflammatory responses and contribute to the development of therapeutic strategies based on endogenous resolution.

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Kinetics of the effect of A2AR engagement on gene expression.Cells were pretreated with CGS 21680 (1 µM) then stimulated as described in Materials and Methods for 30 minutes at 37°C. Values are time-matched ratios of mRNA levels (CGS21680-treated cells/stimulated cells) as determined by real-time PCR, averaged±SEM for three independent experiments performed under identical conditions with a different single donor of cells. *Significantly different from cells stimulated in the absence of CGS 21680.
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pone-0004902-g004: Kinetics of the effect of A2AR engagement on gene expression.Cells were pretreated with CGS 21680 (1 µM) then stimulated as described in Materials and Methods for 30 minutes at 37°C. Values are time-matched ratios of mRNA levels (CGS21680-treated cells/stimulated cells) as determined by real-time PCR, averaged±SEM for three independent experiments performed under identical conditions with a different single donor of cells. *Significantly different from cells stimulated in the absence of CGS 21680.

Mentions: Time-course experiments were undertaken in which cells were stimulated for periods of time ranging from 5 min to 4 h, alone or in presence of the A2AR agonist CGS 21680. Messenger RNA levels for genes of interest were measured by real-time PCR and samples stimulated in the absence or presence of CGS 21680 were compared in a time-matched manner. Depending on the gene, A2AR activation elicited transient (<2 h) or sustained (≥4 h) responses, indicative of gene-specific regulatory processes (Figure 4). However, the impact on gene expression was typically rapid, in most cases becoming apparent in less than 30 minutes.


Impact of anti-inflammatory agents on the gene expression profile of stimulated human neutrophils: unraveling endogenous resolution pathways.

St-Onge M, Dumas A, Michaud A, Laflamme C, Dussault AA, Pouliot M - PLoS ONE (2009)

Kinetics of the effect of A2AR engagement on gene expression.Cells were pretreated with CGS 21680 (1 µM) then stimulated as described in Materials and Methods for 30 minutes at 37°C. Values are time-matched ratios of mRNA levels (CGS21680-treated cells/stimulated cells) as determined by real-time PCR, averaged±SEM for three independent experiments performed under identical conditions with a different single donor of cells. *Significantly different from cells stimulated in the absence of CGS 21680.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2654409&req=5

pone-0004902-g004: Kinetics of the effect of A2AR engagement on gene expression.Cells were pretreated with CGS 21680 (1 µM) then stimulated as described in Materials and Methods for 30 minutes at 37°C. Values are time-matched ratios of mRNA levels (CGS21680-treated cells/stimulated cells) as determined by real-time PCR, averaged±SEM for three independent experiments performed under identical conditions with a different single donor of cells. *Significantly different from cells stimulated in the absence of CGS 21680.
Mentions: Time-course experiments were undertaken in which cells were stimulated for periods of time ranging from 5 min to 4 h, alone or in presence of the A2AR agonist CGS 21680. Messenger RNA levels for genes of interest were measured by real-time PCR and samples stimulated in the absence or presence of CGS 21680 were compared in a time-matched manner. Depending on the gene, A2AR activation elicited transient (<2 h) or sustained (≥4 h) responses, indicative of gene-specific regulatory processes (Figure 4). However, the impact on gene expression was typically rapid, in most cases becoming apparent in less than 30 minutes.

Bottom Line: Genes encoding transcription factors, enzymes and regulatory proteins, as well as secreted cytokines/chemokines showed differential expression.We have identified a set of genes that may be part of important resolution pathways that interfere with cell activation.Identification of these pathways will improve understanding of the capacity of tissues to terminate inflammatory responses and contribute to the development of therapeutic strategies based on endogenous resolution.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy-Physiology, Faculty of Medicine, Centre de Recherche en Rhumatologie et Immunologie du CHUQ, Laval University, Quebec City, Quebec, Canada.

ABSTRACT
Adenosine, prostaglandin E(2), or increased intracellular cyclic AMP concentration each elicit potent anti-inflammatory events in human neutrophils by inhibiting functions such as phagocytosis, superoxide production, adhesion and cytokine release. However, the endogenous molecular pathways mediating these actions are poorly understood. In the present study, we examined their impact on the gene expression profile of stimulated neutrophils. Purified blood neutrophils from healthy donors were stimulated with a cocktail of inflammatory agonists in the presence of at least one of the following anti-inflammatory agents: adenosine A(2A) receptor agonist CGS 21680, prostaglandin E(2), cyclic-AMP-elevating compounds forskolin and RO 20-1724. Total RNA was analyzed using gene chips and real-time PCR. Genes encoding transcription factors, enzymes and regulatory proteins, as well as secreted cytokines/chemokines showed differential expression. We identified 15 genes for which the anti-inflammatory agents altered mRNA levels. The agents affected the expression profile in remarkably similar fashion, suggesting a central mechanism limiting cell activation. We have identified a set of genes that may be part of important resolution pathways that interfere with cell activation. Identification of these pathways will improve understanding of the capacity of tissues to terminate inflammatory responses and contribute to the development of therapeutic strategies based on endogenous resolution.

Show MeSH
Related in: MedlinePlus