Modulation of neutrophil adherence to endothelial cells by platelet-derived adherence-inhibiting factor through interactions with selectin molecules

Abstract

Platelet-derived adherence-inhibiting factor (AIF) has been demonstrated to regulate the neutrophil binding to type IV collagen. In this study, we have examined the effect of AIF on neutrophil adherence to confluently cultured human umbilical vein endothelial cells (EC). AIF inhibited neutrophil adherence to thrombin-or tumor necrosis factor ␣ (TNF-␣)-stimulated EC by 75 or 43%, respectively, but hardly affected neutrophil adherence to resting EC. It is interesting to note that the inhibitory activity of AIF was reduced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation of neutrophils. Pretreatment of neutrophils or EC with AIF inhibited neutrophil adherence to thrombin-or TNF-␣-stimulated EC, suggesting that neutrophils and EC have AIF-binding proteins. Using AIF-Sepharose affinity chromatography, AIF-binding proteins containing L-selectin were isolated from 125 I-labeled resting neutrophils. However, L-selectin was markedly decreased in the AIF-binding fraction from fMLP-stimulated neutrophils. With the use of AIF-affinity chromatography, P-and E-selectins were obtained in the AIF-binding fractions from resting, thrombin-, and TNF-␣-stimulated EC. It is important to note that P-and E-selectin were greatly increased in the AIFbinding fractions from thrombin-and TNF-␣-stimulated EC, respectively. Furthermore, AIF was able to bind to L-selectin-IgG chimeric protein and inhibit the binding of chimeric protein to thrombinor TNF-␣-stimulated EC. In addition, AIF inhibited the binding of anti-P-or anti-E-selectin monoclonal antibody to the lysates of thrombin-or TNF-␣-stimulated EC. Together these observations indicate that AIF could recognize L-, P-, and Eselectins, and modulate neutrophil adherence to EC through interactions with selectin molecules. J.

Short exposure of intestinal epithelial cells to TNF-α and histamine induces Mac-1-mediated neutrophil adhesion independent of protein synthesis

“…Thus, it is tempting to speculate that a similar molecule(s) is involved in Mac-1-mediated neutrophil adhesion to TNF-␣-and histamine-stimulated HT29 cells. It is interesting that cell surface labeling of HT29 cells using 125 I followed by SDS-PAGE analysis [28] preliminarily revealed that proteins with molecular masses of 310, 250, 145, 85, and 65 kDa were up-regulated by stimulation with not only TNF-␣ (5 h and 30 min) but also histamine (30 min; data not shown); however, it is not clear which of these epithelial proteins is actually mediating the interaction with Mac-1 molecule. Epithelial ligand(s) could be identified in the future by techniques such as cross-linking of intestinal epithelial ligand(s) with Mac-1 using chemical cross-linkers, and cloning of intestinal epithelial ligand(s) for Mac-1 from intestinal epithelial cell cDNA library.…”

“…Neutrophils were isolated from sodium citrate-anticoagulated peripheral blood of normal healthy human volunteers with the use of Polymorphprep (Nycomed Pharma AS, Oslo, Norway) centrifugation as described previously [28]. Isolated neutrophils were suspended in buffer A (137 mM NaCl, 2.7 mM KCl, 8.1 mM Na 2 HPO 4 , 1.5 mM KH 2 PO 4 , pH 7.4) at a concentration of 1 ϫ 10 6 cells/mL and used for subsequent experiments.…”

P-selectin mediates reperfusion injury through neutrophil and platelet sequestration in the warm ischemic mouse liver

“…[7][8][9] The leukocyte-endothelial interaction that occurs following ischemia/reperfusion (I/Rp) injury has been shown to be important for microvascular dysfunction and release of cytotoxic mediators such as reactive oxygen intermediates and a variety of proteases. [10][11][12] We 13 and others 5,6,9,14,15 have shown leukocyte accumulation to be a critical determinant of hepatic I/Rp injury. Other blood elements including platelets adhere to the hepatic sinusoids in conditions of ischemia, 16,17 but little is known about their contribution to the injury.…”

Demonstration of the temporal evolution of serum markers of skeletal muscle ischaemia reperfusion injury - An observational clinical study.

“…P-Selectin, a member of the adhesion molecule family expressed in platelets and endothelial cells, is up regulated during IRI and mediates the initial adhesion of leucocytes to the surface of the endothelium, i.e 'leucocyte rolling'. Adhered leucoyctes transmigrate through the endothelium and are activated 6,7 . reported that abrogating TNF-alpha activity with a soluble TNF receptor construct and an anti-TNF-alpha antibody, decreased muscle capillary permeability after IRI in skeletal muscle 12 .…”