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Role of Calprotectin as a Modulator of the IL27-Mediated Proinflammatory Effect on Endothelial Cells.

Dorosz SA, Ginolhac A, Kähne T, Naumann M, Sauter T, Salsmann A, Bueb JL - Mediators Inflamm. (2015)

Bottom Line: However, regarding cytokine IL27, the controversial current knowledge about its inflammatory role and the involved regulatory elements requires clarification.A qPCR-based screening demonstrated high IL27-mediated gene expression of IL7, IL15, CXCL10, and CXCL11.Furthermore, we showed evidence for STAT1 involvement in this process.

View Article: PubMed Central - PubMed

Affiliation: Life Sciences Research Unit, University of Luxembourg, 162a Avenue de la Faïencerie, 1511 Luxembourg City, Luxembourg.

ABSTRACT
An underlying endothelial dysfunction plays a fundamental role in the pathogenesis of cardiovascular events and is the central feature of atherosclerosis. The protein-based communication between leukocytes and inflamed endothelial cells leading to diapedesis has been largely investigated and several key players such as IL6, TNFα, or the damage associated molecular pattern molecule (DAMP) calprotectin are now well identified. However, regarding cytokine IL27, the controversial current knowledge about its inflammatory role and the involved regulatory elements requires clarification. Therefore, we examined the inflammatory impact of IL27 on primary endothelial cells and the potentially modulatory effect of calprotectin on both transcriptome and proteome levels. A qPCR-based screening demonstrated high IL27-mediated gene expression of IL7, IL15, CXCL10, and CXCL11. Calprotectin time-dependent downregulatory effects were observed on IL27-induced IL15 and CXCL10 gene expression. A mass spectrometry-based approach of IL27 ± calprotectin cell stimulation enlightened a calprotectin modulatory role in the expression of 28 proteins, mostly involved in the mechanism of leukocyte transmigration. Furthermore, we showed evidence for STAT1 involvement in this process. Our findings provide new evidence about the IL27-dependent proinflammatory signaling which may be under the control of calprotectin and highlight the need for further investigations on molecules which might have antiatherosclerotic functions.

No MeSH data available.


Related in: MedlinePlus

Effects of IL27, calprotectin, and IL27 + calprotectin cotreatment on STAT1/3 phosphorylation. Relative protein levels of Tyr701-phosphorylated pSTAT1 (a) and Tyr705-pSTAT3 (b) in IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h performed by western blot. Values are represented as mean ± SEM. Normalization was performed against tubulin. Presented statistical significances are between IL27 and IL27 + calprotectin and the indicated p values correspond to ∗∗∗p < 0.001.
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fig7: Effects of IL27, calprotectin, and IL27 + calprotectin cotreatment on STAT1/3 phosphorylation. Relative protein levels of Tyr701-phosphorylated pSTAT1 (a) and Tyr705-pSTAT3 (b) in IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h performed by western blot. Values are represented as mean ± SEM. Normalization was performed against tubulin. Presented statistical significances are between IL27 and IL27 + calprotectin and the indicated p values correspond to ∗∗∗p < 0.001.

Mentions: The signal transducers and activators of transcription (STAT) 1 and 3 are both known to be part of the signaling downstream cascade of IL27 and are activated through phosphorylation [49, 50]. Our proteomic results clearly indicate that calprotectin upregulated IL27-mediated STAT3 protein expression at 6 h whereas protein expression of STAT1 was downregulated at 6 h and upregulated at 24 h (Figure 5). We next analysed by western blotting the phosphorylation levels of STAT1 and STAT3 when HUVECs were stimulated with IL27 (30 ng/mL) ± calprotectin (1 μg/mL) for 3, 6, 12, and 24 h (Figures 7 and S5). Interestingly, the IL27 + calprotectin cotreatment clearly potentialized the IL27-mediated STAT1 phosphorylation at 12 and 24 h but had no effect on the STAT3 phosphorylation over the time, suggesting that calprotectin might act through the regulation of the STAT1 activity to modulate the IL27-dependent proinflammatory signaling.


Role of Calprotectin as a Modulator of the IL27-Mediated Proinflammatory Effect on Endothelial Cells.

Dorosz SA, Ginolhac A, Kähne T, Naumann M, Sauter T, Salsmann A, Bueb JL - Mediators Inflamm. (2015)

Effects of IL27, calprotectin, and IL27 + calprotectin cotreatment on STAT1/3 phosphorylation. Relative protein levels of Tyr701-phosphorylated pSTAT1 (a) and Tyr705-pSTAT3 (b) in IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h performed by western blot. Values are represented as mean ± SEM. Normalization was performed against tubulin. Presented statistical significances are between IL27 and IL27 + calprotectin and the indicated p values correspond to ∗∗∗p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4664814&req=5

fig7: Effects of IL27, calprotectin, and IL27 + calprotectin cotreatment on STAT1/3 phosphorylation. Relative protein levels of Tyr701-phosphorylated pSTAT1 (a) and Tyr705-pSTAT3 (b) in IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h performed by western blot. Values are represented as mean ± SEM. Normalization was performed against tubulin. Presented statistical significances are between IL27 and IL27 + calprotectin and the indicated p values correspond to ∗∗∗p < 0.001.
Mentions: The signal transducers and activators of transcription (STAT) 1 and 3 are both known to be part of the signaling downstream cascade of IL27 and are activated through phosphorylation [49, 50]. Our proteomic results clearly indicate that calprotectin upregulated IL27-mediated STAT3 protein expression at 6 h whereas protein expression of STAT1 was downregulated at 6 h and upregulated at 24 h (Figure 5). We next analysed by western blotting the phosphorylation levels of STAT1 and STAT3 when HUVECs were stimulated with IL27 (30 ng/mL) ± calprotectin (1 μg/mL) for 3, 6, 12, and 24 h (Figures 7 and S5). Interestingly, the IL27 + calprotectin cotreatment clearly potentialized the IL27-mediated STAT1 phosphorylation at 12 and 24 h but had no effect on the STAT3 phosphorylation over the time, suggesting that calprotectin might act through the regulation of the STAT1 activity to modulate the IL27-dependent proinflammatory signaling.

Bottom Line: However, regarding cytokine IL27, the controversial current knowledge about its inflammatory role and the involved regulatory elements requires clarification.A qPCR-based screening demonstrated high IL27-mediated gene expression of IL7, IL15, CXCL10, and CXCL11.Furthermore, we showed evidence for STAT1 involvement in this process.

View Article: PubMed Central - PubMed

Affiliation: Life Sciences Research Unit, University of Luxembourg, 162a Avenue de la Faïencerie, 1511 Luxembourg City, Luxembourg.

ABSTRACT
An underlying endothelial dysfunction plays a fundamental role in the pathogenesis of cardiovascular events and is the central feature of atherosclerosis. The protein-based communication between leukocytes and inflamed endothelial cells leading to diapedesis has been largely investigated and several key players such as IL6, TNFα, or the damage associated molecular pattern molecule (DAMP) calprotectin are now well identified. However, regarding cytokine IL27, the controversial current knowledge about its inflammatory role and the involved regulatory elements requires clarification. Therefore, we examined the inflammatory impact of IL27 on primary endothelial cells and the potentially modulatory effect of calprotectin on both transcriptome and proteome levels. A qPCR-based screening demonstrated high IL27-mediated gene expression of IL7, IL15, CXCL10, and CXCL11. Calprotectin time-dependent downregulatory effects were observed on IL27-induced IL15 and CXCL10 gene expression. A mass spectrometry-based approach of IL27 ± calprotectin cell stimulation enlightened a calprotectin modulatory role in the expression of 28 proteins, mostly involved in the mechanism of leukocyte transmigration. Furthermore, we showed evidence for STAT1 involvement in this process. Our findings provide new evidence about the IL27-dependent proinflammatory signaling which may be under the control of calprotectin and highlight the need for further investigations on molecules which might have antiatherosclerotic functions.

No MeSH data available.


Related in: MedlinePlus