Limits...
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 gene expression. Relative mRNA levels of IL7, IL15, CXCL10, and CXCL11 of IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h are represented as mean ± SEM (n = 6). Indicated p values are corresponding to significant differences between IL27 and IL27 + calprotectin: ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4664814&req=5

fig3: Effects of IL27, calprotectin, and IL27/calprotectin cotreatment on gene expression. Relative mRNA levels of IL7, IL15, CXCL10, and CXCL11 of IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h are represented as mean ± SEM (n = 6). Indicated p values are corresponding to significant differences between IL27 and IL27 + calprotectin: ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Mentions: To evaluate the possible role of calprotectin in the regulation of the expression of the IL27-dependent upregulated genes IL7, IL15, CXCL10, and CXCL11, HUVECs were treated with IL27 (30 ng/mL) ± calprotectin (1 μg/mL) and the relative gene expression was determined by RT-qPCR for the different time points 3, 6, 12, and 24 h as shown in Figure 3. Calprotectin induced downregulation of IL27-mediated gene expression of IL15 and CXCL10 while no significant effect was observed neither on IL7 nor on CXCL11. Interestingly, calprotectin decreased IL27-induced gene expression of IL15 by half at all time points, whereas its downregulating effect on CXCL10 was weaker and limited to the early time points 3 h and 6 h. These results point to a specific downregulatory role of calprotectin in the endothelial IL27-dependent signaling leading to gene expression of IL15 and CXCL10.


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 gene expression. Relative mRNA levels of IL7, IL15, CXCL10, and CXCL11 of IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h are represented as mean ± SEM (n = 6). Indicated p values are corresponding to significant differences between IL27 and IL27 + calprotectin: ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: Effects of IL27, calprotectin, and IL27/calprotectin cotreatment on gene expression. Relative mRNA levels of IL7, IL15, CXCL10, and CXCL11 of IL27 (30 ng/mL) ± calprotectin (1 μg/mL)-stimulated HUVECs for 3, 6, 12, and 24 h are represented as mean ± SEM (n = 6). Indicated p values are corresponding to significant differences between IL27 and IL27 + calprotectin: ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
Mentions: To evaluate the possible role of calprotectin in the regulation of the expression of the IL27-dependent upregulated genes IL7, IL15, CXCL10, and CXCL11, HUVECs were treated with IL27 (30 ng/mL) ± calprotectin (1 μg/mL) and the relative gene expression was determined by RT-qPCR for the different time points 3, 6, 12, and 24 h as shown in Figure 3. Calprotectin induced downregulation of IL27-mediated gene expression of IL15 and CXCL10 while no significant effect was observed neither on IL7 nor on CXCL11. Interestingly, calprotectin decreased IL27-induced gene expression of IL15 by half at all time points, whereas its downregulating effect on CXCL10 was weaker and limited to the early time points 3 h and 6 h. These results point to a specific downregulatory role of calprotectin in the endothelial IL27-dependent signaling leading to gene expression of IL15 and CXCL10.

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