scholarly journals The Fcγ Receptor–Mediated Respiratory Burst of Rolling Neutrophils to Cytokine-Activated, Immune Complex-Bearing Endothelial Cells Depends on L-Selectin But Not on E-Selectin

Blood ◽  
1998 ◽  
Vol 91 (7) ◽  
pp. 2558-2564 ◽  
Author(s):  
Dieter Fröhlich ◽  
Olivier Spertini ◽  
René Moser
Keyword(s):  
Endothelial Cells ◽  
Immune Complex ◽  
Respiratory Burst ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Auxiliary Function ◽  
H2o2 Production ◽  
H2o2 Generation ◽  
Umbilical Vein Endothelial Cells ◽  
Static Conditions

Abstract Intracellular H2O2 generation, as a measure of the respiratory burst, was determined after stimulation of neutrophils by immune complex (IC)-bearing human umbilical vein endothelial cells. Under static conditions, neutrophils basically responded to the immune deposits on resting endothelial cells. The rotating shear forces of ≈0.7 dynes/cm2, corresponding to the physiological flow in postcapillary venules, completely abolished this basal H2O2 generation. After activation of the IC-bearing endothelial layers with interleukin-1 (IL-1) or tumor necrosis factor (TNF), or both, for 4 hours, rolling adhesion of the neutrophils was induced, accompanied by considerable H2O2 production. The neutrophil respiratory burst was prominently inhibited by anti-FcγRIII MoAb 3G8 (72.4%), and partially by MoAb 2E1 against FcγRII (38.5%). Both MoAbs together inhibited the Fc-mediated H2O2generation by 93.4%. The respiratory burst and rolling adhesion were markedly blocked by MoAb LAM1-3 against L-selectin (91.3%), whereas the nonfunctional anti-L-selectin MoAb LAM1-14 was ineffective. F(ab)2′ fragments of MoAb 7A9 against E-selectin inhibited neutrophil rolling by 98.6%, but not the respiratory burst. Moreover, rolling adhesion of neutrophils and the related oxidative burst were CD11b/CD18- independent. In summary, L-selectin has a unique auxiliary function in triggering the FcγR-mediated respiratory burst of rolling neutrophils to IC-bearing endothelial cells, thereby substituting CD11b/CD18 under conditions of flow.

scholarly journals The Fcγ Receptor–Mediated Respiratory Burst of Rolling Neutrophils to Cytokine-Activated, Immune Complex-Bearing Endothelial Cells Depends on L-Selectin But Not on E-Selectin

Blood ◽  
1998 ◽  
Vol 91 (7) ◽  
pp. 2558-2564 ◽  
Author(s):  
Dieter Fröhlich ◽  
Olivier Spertini ◽  
René Moser
Keyword(s):  
Endothelial Cells ◽  
Immune Complex ◽  
Respiratory Burst ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Auxiliary Function ◽  
H2o2 Production ◽  
H2o2 Generation ◽  
Umbilical Vein Endothelial Cells ◽  
Static Conditions

Intracellular H2O2 generation, as a measure of the respiratory burst, was determined after stimulation of neutrophils by immune complex (IC)-bearing human umbilical vein endothelial cells. Under static conditions, neutrophils basically responded to the immune deposits on resting endothelial cells. The rotating shear forces of ≈0.7 dynes/cm2, corresponding to the physiological flow in postcapillary venules, completely abolished this basal H2O2 generation. After activation of the IC-bearing endothelial layers with interleukin-1 (IL-1) or tumor necrosis factor (TNF), or both, for 4 hours, rolling adhesion of the neutrophils was induced, accompanied by considerable H2O2 production. The neutrophil respiratory burst was prominently inhibited by anti-FcγRIII MoAb 3G8 (72.4%), and partially by MoAb 2E1 against FcγRII (38.5%). Both MoAbs together inhibited the Fc-mediated H2O2generation by 93.4%. The respiratory burst and rolling adhesion were markedly blocked by MoAb LAM1-3 against L-selectin (91.3%), whereas the nonfunctional anti-L-selectin MoAb LAM1-14 was ineffective. F(ab)2′ fragments of MoAb 7A9 against E-selectin inhibited neutrophil rolling by 98.6%, but not the respiratory burst. Moreover, rolling adhesion of neutrophils and the related oxidative burst were CD11b/CD18- independent. In summary, L-selectin has a unique auxiliary function in triggering the FcγR-mediated respiratory burst of rolling neutrophils to IC-bearing endothelial cells, thereby substituting CD11b/CD18 under conditions of flow.

Syndecan-4-dependent signaling in the inhibition of endotoxin-induced endothelial adherence of neutrophils by antithrombin

2003 ◽  
Vol 90 (12) ◽  
pp. 1150-1157 ◽  
Author(s):  
Nicole Kaneider ◽  
Ellen Förster ◽  
Birgit Mosheimer ◽  
Daniel Sturn ◽  
Christian Wiedermann
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Human Neutrophils ◽  
Heparin Binding ◽  
Clotting Factors ◽  
Edema Formation ◽  
Granulocyte Elastase ◽  
Umbilical Vein Endothelial Cells ◽  
Static Conditions

SummaryCirculating endotoxin is elevated in sepsis and plays a role in endothelial dysfunction whereas antithrombin is decreased by virtue of its consumption during complex formation with clotting factors and by proteolytic degradation by granulocyte elastase. Dysfunction of endothelium results in enhanced leukocyte rolling and diapedesis into tissues leading to edema formation and injury. Antithrombin exerts beneficial effects on endothelial function in sepsis. A direct anti-inflammatory action of anti-thrombin in inflammatory cells is exerted via heparan sulfate proteoglycans. In this study, we investigated whether antithrom-bin affects endotoxin-induced adhesion of neutrophils to human endothelial cells in vitro and whether glycosaminoglycans are involved in its signaling. Adhesion of human neutrophils to monolayers of umbilical vein endothelial cells was tested under static conditions. Endothelial cells were pretreated with endotoxin, interleukin-1, heparinase-I, chondroitinase-ABC or anti-syndecan-4-antibody. Endotoxin and interleukin-1 increased neutrophil adherence to human umbilical vein endothelial cells which was inhibited by antithrombin. Concomitant incubation with pentasaccharide abolished this effect of antithrombin. Treatment of endothelial cells with heparinase or chondroitinase led to higher adhesion and prevented effects of antithrom-bin. With antibodies to syndecan-4, enhanced adhesion of neutrophils was observed. As studied by Western blotting, endo-toxin-induced signaling was diminished by antithrombin and the effect was reversible by chondroitinase or heparinase. From our results, we can conclude that endotoxin-induced adhesion of leukocytes to endothelium can be reversed by ligation of syndecan-4 with antithrombin´s heparin-binding site and interferences with stress response signaling events in endothelium.

scholarly journals Interleukin-1 Enhances the Ability of Cultured Human Umbilical Vein Endothelial Cells to Oxidize Linoleic Acid

1995 ◽  
Vol 270 (29) ◽  
pp. 17279-17286 ◽  
Author(s):  
Mercedes Camacho ◽  
Nuria Godessart ◽  
Rosa Antón ◽  
Montserrat Garca ◽  
Lus Vila
Keyword(s):  
Endothelial Cells ◽  
Linoleic Acid ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Bacterial adherence to human endothelial cells

1988 ◽  
Vol 65 (3) ◽  
pp. 1372-1376 ◽  
Author(s):  
P. D. Thomas ◽  
F. W. Hampson ◽  
G. W. Hunninghake
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Bacterial Adherence ◽  
Pulmonary Infections ◽  
Human Endothelial Cells ◽  
Cell Surface Antigens ◽  
Lung Endothelium ◽  
Umbilical Vein Endothelial Cells ◽  
Reduced Temperature

The adult respiratory distress syndrome (ARDS) is frequently caused by exposure of the lung endothelium to circulating endotoxin (lipopolysaccharide, LPS) and pulmonary infections frequently develop during the course of ARDS. The present studies demonstrate that LPS and interleukin 1 (IL-1, a mediator released by endothelial cells after exposure to LPS) enhance the adherence of Staphylococcus aureus to human umbilical vein endothelial cells. gamma-Interferon, another mediator that induces expression of some cell surface antigens on endothelial cells, had no effect on bacterial adherence. The adherence of bacteria to endothelium was increased by prior opsonization of the bacteria with fresh human serum and was reduced by prior absorption of the serum with bacteria before the use of the serum for opsonization. The capacity of LPS to increase bacterial adherence was time dependent and was maximally expressed after 6 h of exposure; it was blocked by exposure of endothelial cells to LPS in the presence of reduced temperature or dactinomycin (Actinomycin D). These observations suggest that circulating LPS not only can trigger the development of ARDS but also may predispose the lung to the development of pulmonary infections by increasing adherence of bacteria to endothelium.

THE EFFECT OF PHORBOL DIESTERS AND INTERLEUKINS ON THE SECRETION OF VON WILLEBRAND FACTOR BY HUMAN ENDOTHELIAL CELLS IN CULTURE

1987 ◽  
Author(s):  
J C Giddings ◽  
L Shall
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Interleukin 2 ◽  
Morphological Evidence ◽  
Adhesive Properties ◽  
Umbilical Vein Endothelial Cells ◽  
Von Willebrand ◽  
Willebrand Factor

Human umbilical vein endothelial cells (EC) were cultured in the presence of 4p-phorbol 12-myristate 13-acetate (PMA, 10ug/l), interleukin 1 (IL-1, 1 unit/ml) and interleukin 2 (IL-2, 1 unit/ml), and secretion of von Willebrand factor activity (vWF, Ristocetin co-factor) and von Willebrand factor antigen (vWFAG, ELISA Technique) measured at intervals. Confluent control EC were treated with PMA, IL-1 and IL-2, and the supernatant medium assayed for release of vWF and vWFAg. Treated cells were also examined for vWFAg by immuno-fluorescence. The levels of both vWF and vWFAg in cultures containing IL-1 were significantly higher than those in control cultures after 5-6 days growth. Moreover, vWF and vWFAg increased significantly in the supernatant of confluent control EC incubated further in the presence of IL-1. Furthermore, the characteristic fluorescence pattern of endothelial vWFAg was markedly reduced in EC treated with IL-1. The levels of vWF and vWFAg in cultures containing PMA were also significantly higher than those of control cultures. In these conditions, however, the growth of cells appeared to be enhanced, and confluence was observed after about 6 days in the presence of PMA compared to 9 - 10 days in control cultures. The mean levels of vWF and vWFAg in the supernatant of EC incubated with PMA were higher than the control values but the differences were not statistically significant. Immunofluorescence of PMA-treated cells suggested that vWFAg might be less granular than in control cells but the differences were not as marked as those seen with IL-1. The results of all assays in the presence of IL-2 were not significantly different from those of control cells. In all instances no morphological evidence of endothelial injury was observed and more than 90% of cells remained viable at the termination of cultures. The results indicated that the synthesis and release of vWF were increased in the presence of PMA, and secretion of vWF was stimulated by IL-1. The data suggest that secreted vWF might contribute to the previously reported enhanced procoagulant and adhesive properties of EC treated with these substances.

scholarly journals Shear-dependent Eosinophil Transmigration on Interleukin 4–stimulated Endothelial Cells

2001 ◽  
Vol 194 (12) ◽  
pp. 1699-1709 ◽  
Author(s):  
Susan L. Cuvelier ◽  
Kamala D. Patel
Keyword(s):  
Endothelial Cells ◽  
Pertussis Toxin ◽  
Umbilical Vein ◽  
Interleukin 4 ◽  
Western Blots ◽  
Surface Pretreatment ◽  
Endothelial Cell Surface ◽  
Umbilical Vein Endothelial Cells ◽  
Static Conditions

Leukocyte infiltration into inflammatory sites is regulated by the expression of adhesion and activation proteins, yet the role of these proteins in shear-dependent transmigration is poorly understood. We examined eosinophil recruitment on cytokine-stimulated human umbilical vein endothelial cells (HUVECs) under laminar flow conditions. Eosinophils rapidly transmigrated on interleukin (IL)-4–, but not TNF-stimulated HUVECs. Transmigration was shear dependent, with up to 90% of eosinophils transmigrating in the presence of shear and less than 25% of cells transmigrating under static conditions. Eosinophils express CC chemokine receptor CCR3 and are responsive to various CC chemokines. The effects of chemokines are mediated primarily through Gαi, which is pertussis toxin sensitive. Greater than 65% of shear-dependent eosinophil transmigration on IL-4–stimulated HUVECs was blocked by either pertussis toxin or by an anti-CCR3 monoclonal antibody. Using reverse transcription polymerase chain reaction (RT-PCR) and Western blots, we found that IL-4–stimulated HUVECs produce both mRNA and protein for eotaxin-3. Eotaxin-3 was both released by HUVECs and expressed on the endothelial cell surface. Pretreatment of HUVECs with an anti–eotaxin-3 antibody blocked eosinophil transmigration to the same extent as an anti-CCR3 antibody. These results indicate that IL-4–stimulated HUVECs support shear-dependent eosinophil transmigration by upregulating eotaxin-3, and that surface association is critical for the role of eotaxin-3 in transmigration.

Induction of Tissue Factor Synthesis in Human Umbilical Vein Endothelial Cells Involves Protein Kinase C

1992 ◽  
Vol 67 (04) ◽  
pp. 473-477 ◽  
Author(s):  
Kjell Sverre Pettersen ◽  
Merete Thune Wiiger ◽  
Nobuhiro Narahara ◽  
Kiyoshi Andoh ◽  
Gustav Gaudernack ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Tissue Factor ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Interleukin 1Β ◽  
Human Umbilical Vein ◽  
Kinase C ◽  
Umbilical Vein Endothelial Cells

SummaryIncubation of human umbilical vein endothelial cells with one of the following compounds: endotoxin, recombinant interleukin-1β, recombinant tumor necrosis factor α, allogenic lymphocyte subpopulations or phorbol ester resulted in significant induction of tissue factor synthesis. Diacylglycerol had the same effect and also enhanced synergistically the induction caused by endotoxin and interleukin-1β. Two different inhibitors of protein kinase C, H7 and sphingosine, inhibited tissue factor synthesis at concentrations which did not depress protein synthesis in general, suggesting that protein kinase C is involved in the processes leading to tissue factor synthesis. Cells down-regulated for the tissue factor response to TPA responded essentially normally to endotoxin and interleukin-1 with regard to tissue factor synthesis.

PAF mediates neutrophil adhesion to thrombin or TNF-stimulated endothelial cells under shear stress

1995 ◽  
Vol 269 (1) ◽  
pp. C42-C47 ◽  
Author(s):  
D. Macconi ◽  
M. Foppolo ◽  
S. Paris ◽  
M. Noris ◽  
S. Aiello ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Platelet Activating Factor ◽  
Flow Chamber ◽  
Tnf Alpha ◽  
Image Analysis System ◽  
Flow Conditions ◽  
Necrosis Factor Alpha ◽  
Umbilical Vein Endothelial Cells ◽  
Static Conditions

Platelet-activating factor (PAF) is known to modulate polymorphonuclear leukocyte (PMN) adhesion to endothelial cells cultured under static conditions and activated by thrombin. In contrast, there are no data on the role of PAF in PMN adhesion to cells exposed to flow conditions and activated by stimuli other than thrombin. Here we used the PAF receptor antagonist L-659,989 to evaluate PMN adhesion to human umbilical vein endothelial cells (HUVEC) in basal conditions or upon challenge with thrombin or tumor necrosis factor-alpha (TNF-alpha). Experiments were performed under dynamic flow using a parallel-plate flow chamber and a computer-based image analysis system. Rolling and adhesion of PMNs to endothelial cells significantly increased upon stimulation with thrombin. Thrombin-stimulated HUVEC also synthesized higher amounts of PAF than untreated cells. Pretreatment of PMNs with L-659,989 significantly reduced their rolling and adhesion to thrombin-activated HUVEC. Stimulation of HUVEC with TNF-alpha significantly increased the number of rolling and adherent PMNs as compared with untreated cells. Adhesion of PMNs to and migration across TNF-alpha-stimulated HUVEC were reduced by L-659,989, whereas cell rolling was unchanged. We conclude that PAF mediates leukocyte interaction under flow conditions with HUVEC activated by inflammatory stimuli.

scholarly journals Interleukin-1 induces interleukin-8 secretion from endothelial cells by a juxtacrine mechanism

Blood ◽  
1994 ◽  
Vol 84 (12) ◽  
pp. 4242-4248 ◽  
Author(s):  
G Kaplanski ◽  
C Farnarier ◽  
S Kaplanski ◽  
R Porat ◽  
L Shapiro ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Mannose Receptor ◽  
Interleukin 8 ◽  
Facs Analysis ◽  
Peripheral Blood Mononuclear ◽  
Umbilical Vein Endothelial Cells

Inflammation is characterized by migration of neutrophils through the endothelium, and the chemokine interleukin-8 (IL-8) appears to be involved. We asked whether adherence of cells bearing a membrane-form of interleukin 1 (IL-1) induces IL-8 secretion from human umbilical vein endothelial cells (HUVEC) and fibroblasts. Human peripheral blood mononuclear cells (PBMC) were stimulated with endotoxin for 12 hours and then fixed for 4 hours with paraformaldehyde. When these cells were added to HUVEC or fibroblasts, IL-8 production was induced. This stimulation by fixed PBMC was attributed to IL-1, because pretreatment of HUVEC or fibroblasts with IL-1 receptor antagonist (IL-1Ra) reduced the induction by 95% and 80%, respectively, P < .005. Using anti-IL-1 alpha monoclonal antibodies, reduction was complete, whereas anti-IL-1 beta had no effect. IL-1 alpha was shown on the surface of monocytes by fluorescence-activated cell sorter (FACS) analysis. Blockade of IL-1 receptors on PBMC did not affect the activity of membrane-associated IL- 1 alpha, indicating that IL-1 is not anchored to the membrane through its receptors. However, PBMC treated with D-mannose before fixation resulted in a loss of activity; this loss of activity was associated with release of IL-1 alpha, not IL-1 beta, into the supernatant. Thus, anchoring of IL-1 alpha to the membrane may be via a lectin or mannose receptor-like interaction. Blockade of membrane IL-1 alpha required a 30-fold and fivefold excess of IL-1Ra compared with the amount required to block soluble IL-1 beta and IL-1 alpha, respectively. We conclude that the fixed PBMC IL-8 inducing activity is almost entirely caused by IL-1, that this represents IL-1 alpha bound to a surface lectin or mannose receptor on the monocyte, and that it functions in inflammation via juxtacrine interactions.

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