STAT-6-Mediated Control of P-Selectin by Substance P and Interleukin-4 in Human Dermal Endothelial Cells

Abstract

P-Selectin expressed on endothelial cells contributes to acute and chronic inflammation by promoting leukocyte tethering/rolling. Despite increasing evidence of P-selectin expression on human umbilical vein endothelial cells in vitro , the regulatory mechanisms of P-selectin expression on dermal endothelial cells in skin diseases are not fully understood. Here , we demonstrate increased expression of P-selectin in dermal vessels of regional skin in urticaria and atopic dermatitis. The present in vitro analyses with human dermal microvascular endothelial cells (HDMECs) revealed that histamine rapidly induced P-selectin expression. Interleukin (IL)-4 and IL-13 induced prolonged expression of surface P-selectin by HDMECs. A combination of tumor necrosis factor-␣ and IL-4 inhibited P-selectin expression. Pretreatment of HDMECs with tumor necrosis factor-␣ followed by incubation with IL-4 markedly increased P-selectin expression. Notably , incubation with substance P alone induced prolonged P-selectin expression. Activation of STAT6 appears to be a key factor in P-selectin expression induced by substance P and IL-4 because treatment with STAT6 decoy oligodeoxynucleotides significantly inhibited P-selectin expression. The present results indicate that novel , complex mechanisms are involved in endothelial P-selectin expression in the skin. STAT6 in dermal endothelial cells appears to be a potent target for controlling cellular infiltrate in allergic and/or neuroinflammatory skin diseases. Selectins expressed by endothelial cells interact with leukocyte counterreceptors and mediate the initial adhesion of leukocytes and their rolling along endothelial surfaces. Several lines of evidences indicate that endothelial selectins play important roles in inflammation. Mice lacking E-, P-, and/or L-selectins exhibit reduced leukocyte rolling and reduced extravasation of neutrophils in thioglycolate-induced peritonitis.1 P-and E-selectins are also essential for the development of irritation dermatitis and allergic contact hypersensitivity.2 Previous studies indicate that eosinophils have marked avidity for P-selectin, but not E-selectin.3-5 In mice, P-selectin deficiency causes reduced eosinophil rolling and reduced tissue eosinophilia in rag-weed peritonitis.

Genome-Wide Approaches Reveal Functional Interleukin-4-Inducible STAT6 Binding to the Vascular Cell Adhesion Molecule 1 Promoter

“…Previous studies have shown that IL-4 activates STAT6 in endothelial cells (36,40). Moreover, STAT6 has been implicated in IL-4-mediated induction of P-selectin and eotaxin-3 in endothelial cells (18,20,32). As shown in Fig.…”

“…Activation of the receptor results in Janus kinase 1/2 (JAK-1/2)-dependent tyrosine phosphorylation and subsequent dimerization of signal transducer and activation of transcription 6 (STAT6), which then translocates to the nucleus and binds to consensus sequences (TTCN 3-4 GAA) found within promoters of IL-4-regulated target genes (14,27). Previous studies with endothelial cells have demonstrated that IL-4 induces the expression of CXCL-8, inducible nitric oxide synthase (iNOS) (15), urokinase-type plasminogen activator (u-PA) (46), vascular endothelial growth factor (VEGF) (15), P-selectin (20,32,47), monocyte chemoattractant protein 1 (MCP-1) (39), CCL26 (18), , 15-lipoxygenase (24), and osteoprotegerin (41). In addition, previous studies have shown that IL-4 upregulates the expression of VCAM-1 in endothelial cells (4,10,14,23,26,37,40).…”

A Mouse Model of Airway Disease: Oncostatin M-Induced Pulmonary Eosinophilia, Goblet Cell Hyperplasia, and Airway Hyperresponsiveness Are STAT6 Dependent, and Interstitial Pulmonary Fibrosis Is STAT6 Independent

“…Upregulation of P-selectin and VCAM-1 has been documented following OSM (71,72) or IL-4 and IL-13 (73-75) stimulation of vascular endothelial cells. Because STAT6 is necessary for P-selectin (73,74), but not VCAM-1 (15,32,76) expression, this suggests that abrogation of eosinophil accumulation into STAT6 2/2 mouse lungs following AdmOSM administration could be attributed to the inhibition of P-selectin upregulation in concert with abolished eotaxin-1/2 expression. Neutrophils also use P-selectin glycoprotein ligand 1-P-selectin interactions during cell trafficking (68), and their chemotaxis is mediated by the neutrophil-selective CXC chemokine KC (77).…”

Focal adhesion kinase-related nonkinase (FRNK) negatively regulates IL-4-mediated inflammation

“…IL-4 is an important cytokine in models of allergic disease and asthma (Grünig et al, 1998;Luzina et al, 2012;Pelaia et al, 2012;Ricci et al, 1997). In endothelial cells IL-4 exerts its effects through the IL-4 type II receptor, IL13RA1 (Palmer-Crocker et al, 1996;Schnyder et al, 1996), resulting in the expression of STAT6-responsive proteins including those involved in eosinophil recruitment (Cuvelier & Patel, 2001;Hoeck and Woisetschlager, 2001;Miyazaki et al, 2006;Patel, 1998;Tozawa et al, 2011). Although STAT6 is crucial for VCAM-1 and CCL26 expression, FRNK had no effect on STAT6 phosphorylation, nuclear localization or total protein expression (Fig.…”

“…In endothelial cells, IL-4 acts solely through the IL-4 type II receptor (Schnyder et al, 1996), leading to increased phosphorylation of JAK2 and STAT6 (Palmer-Crocker et al, 1996). Upon phosphorylation, STAT6 translocates into the nucleus where it regulates the expression of over 900 genes including those encoding vascular cell adhesion molecule 1 (VCAM-1) (Hopkins, 2013), CCL26 (Tozawa et al, 2011) and Pselectin (Miyazaki et al, 2006;Yao et al, 1996). VCAM-1 and Pselectin then mediate the recruitment of eosinophils, lymphocytes and monocytes (Matsumoto et al, 1997;Patel, 1999), and CCL26 activates eosinophils and supports eosinophil transmigration (Cuvelier and Patel, 2001).…”