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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

Venn diagrams on IL27- and calprotectin-stimulated HUVECs including significant regulated genes with p value p < 0.05: red: upregulated, green: downregulated, and black: up- and downregulation different in compared concentrations. (a) Venn diagram: HUVECs stimulated with IL27 (10, 30, and 100 ng/mL). (b) Venn diagram: HUVECs stimulated with calprotectin (1, 5, and 10 μg/mL). ACE, angiotensin-converting enzyme; BCL2, B-cell lymphoma 2; BCL2L1, BCL2-like 1; C3, complement component 3; CD, cluster of differentiation; CSF, colony stimulating factor; CCL2/MCP-1, monocyte chemotactic protein-1; CCL5/RANTES, regulated on activation, normal T cell expressed and secreted; CXCL10, interferon gamma-induced protein 10; CXCL11, interferon-inducible T cell alpha chemoattractant; CYP7A1, cholesterol 7 alpha-hydroxylase; FAS, Fas cell surface death receptor; ICAM1, intercellular adhesion molecule 1; IKBKB, inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta; IL, interleukin; LTA, lymphotoxin-α; NFKB2, nuclear factor-kappa-B p100 subunit; NOS2A, inducible nitric oxide synthase; PGK1, phosphoglycerate kinase 1; PTGS2, prostaglandin G/H synthase and cyclooxygenase; SELE, selectin E; SELP, selectin P; SMAD3, mothers against decapentaplegic homolog 3; STAT, signal transducers and activators of transcription.
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fig1: Venn diagrams on IL27- and calprotectin-stimulated HUVECs including significant regulated genes with p value p < 0.05: red: upregulated, green: downregulated, and black: up- and downregulation different in compared concentrations. (a) Venn diagram: HUVECs stimulated with IL27 (10, 30, and 100 ng/mL). (b) Venn diagram: HUVECs stimulated with calprotectin (1, 5, and 10 μg/mL). ACE, angiotensin-converting enzyme; BCL2, B-cell lymphoma 2; BCL2L1, BCL2-like 1; C3, complement component 3; CD, cluster of differentiation; CSF, colony stimulating factor; CCL2/MCP-1, monocyte chemotactic protein-1; CCL5/RANTES, regulated on activation, normal T cell expressed and secreted; CXCL10, interferon gamma-induced protein 10; CXCL11, interferon-inducible T cell alpha chemoattractant; CYP7A1, cholesterol 7 alpha-hydroxylase; FAS, Fas cell surface death receptor; ICAM1, intercellular adhesion molecule 1; IKBKB, inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta; IL, interleukin; LTA, lymphotoxin-α; NFKB2, nuclear factor-kappa-B p100 subunit; NOS2A, inducible nitric oxide synthase; PGK1, phosphoglycerate kinase 1; PTGS2, prostaglandin G/H synthase and cyclooxygenase; SELE, selectin E; SELP, selectin P; SMAD3, mothers against decapentaplegic homolog 3; STAT, signal transducers and activators of transcription.

Mentions: To examine the effect of IL27 and calprotectin on endothelial cell gene expression, a multiplex gene array analysis of 96 genes including cytokines, growth factors, and other immune response genes was performed. In order to optimize the stimulation concentrations for IL27 and calprotectin, 3 different concentrations for IL27 (10, 30, and 100 ng/mL) and for calprotectin (1, 5, and 10 μg/mL) were used, according to published data [19, 33, 48]. The relationship of significant differentially expressed genes following stimulation with IL27 is shown in Figure 1(a). Nineteen unique significantly expressed genes were found among which 15 were upregulated and 4 were downregulated. Interestingly, the highest upregulations were observed for IL7, IL15, CXCL10, and CXCL11 (Table S1). Regarding calprotectin cell stimulation, 15 unique significantly expressed genes were found, among which 11 genes were upregulated and 4 genes were downregulated, with the gene PTGS2 being upregulated after 1 and 10 μg/mL calprotectin treatment and downregulated after 5 μg/mL calprotectin stimulation. The highest differential gene expression was observed for IL7 and CCL5 (Table S1).


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)

Venn diagrams on IL27- and calprotectin-stimulated HUVECs including significant regulated genes with p value p < 0.05: red: upregulated, green: downregulated, and black: up- and downregulation different in compared concentrations. (a) Venn diagram: HUVECs stimulated with IL27 (10, 30, and 100 ng/mL). (b) Venn diagram: HUVECs stimulated with calprotectin (1, 5, and 10 μg/mL). ACE, angiotensin-converting enzyme; BCL2, B-cell lymphoma 2; BCL2L1, BCL2-like 1; C3, complement component 3; CD, cluster of differentiation; CSF, colony stimulating factor; CCL2/MCP-1, monocyte chemotactic protein-1; CCL5/RANTES, regulated on activation, normal T cell expressed and secreted; CXCL10, interferon gamma-induced protein 10; CXCL11, interferon-inducible T cell alpha chemoattractant; CYP7A1, cholesterol 7 alpha-hydroxylase; FAS, Fas cell surface death receptor; ICAM1, intercellular adhesion molecule 1; IKBKB, inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta; IL, interleukin; LTA, lymphotoxin-α; NFKB2, nuclear factor-kappa-B p100 subunit; NOS2A, inducible nitric oxide synthase; PGK1, phosphoglycerate kinase 1; PTGS2, prostaglandin G/H synthase and cyclooxygenase; SELE, selectin E; SELP, selectin P; SMAD3, mothers against decapentaplegic homolog 3; STAT, signal transducers and activators of transcription.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4664814&req=5

fig1: Venn diagrams on IL27- and calprotectin-stimulated HUVECs including significant regulated genes with p value p < 0.05: red: upregulated, green: downregulated, and black: up- and downregulation different in compared concentrations. (a) Venn diagram: HUVECs stimulated with IL27 (10, 30, and 100 ng/mL). (b) Venn diagram: HUVECs stimulated with calprotectin (1, 5, and 10 μg/mL). ACE, angiotensin-converting enzyme; BCL2, B-cell lymphoma 2; BCL2L1, BCL2-like 1; C3, complement component 3; CD, cluster of differentiation; CSF, colony stimulating factor; CCL2/MCP-1, monocyte chemotactic protein-1; CCL5/RANTES, regulated on activation, normal T cell expressed and secreted; CXCL10, interferon gamma-induced protein 10; CXCL11, interferon-inducible T cell alpha chemoattractant; CYP7A1, cholesterol 7 alpha-hydroxylase; FAS, Fas cell surface death receptor; ICAM1, intercellular adhesion molecule 1; IKBKB, inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta; IL, interleukin; LTA, lymphotoxin-α; NFKB2, nuclear factor-kappa-B p100 subunit; NOS2A, inducible nitric oxide synthase; PGK1, phosphoglycerate kinase 1; PTGS2, prostaglandin G/H synthase and cyclooxygenase; SELE, selectin E; SELP, selectin P; SMAD3, mothers against decapentaplegic homolog 3; STAT, signal transducers and activators of transcription.
Mentions: To examine the effect of IL27 and calprotectin on endothelial cell gene expression, a multiplex gene array analysis of 96 genes including cytokines, growth factors, and other immune response genes was performed. In order to optimize the stimulation concentrations for IL27 and calprotectin, 3 different concentrations for IL27 (10, 30, and 100 ng/mL) and for calprotectin (1, 5, and 10 μg/mL) were used, according to published data [19, 33, 48]. The relationship of significant differentially expressed genes following stimulation with IL27 is shown in Figure 1(a). Nineteen unique significantly expressed genes were found among which 15 were upregulated and 4 were downregulated. Interestingly, the highest upregulations were observed for IL7, IL15, CXCL10, and CXCL11 (Table S1). Regarding calprotectin cell stimulation, 15 unique significantly expressed genes were found, among which 11 genes were upregulated and 4 genes were downregulated, with the gene PTGS2 being upregulated after 1 and 10 μg/mL calprotectin treatment and downregulated after 5 μg/mL calprotectin stimulation. The highest differential gene expression was observed for IL7 and CCL5 (Table S1).

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