Single membrane tether extraction from adult and neonatal dermal microvascular endothelial cells

2007 ◽  
Vol 292 (4) ◽  
pp. C1272-C1279 ◽  
Author(s):  
Yong Chen ◽  
Gaurav Girdhar ◽  
Jin-Yu Shao
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Cell Lineage ◽  
Constitutive Relationship ◽  
Cell Surface Receptor ◽  
Microvascular Endothelial Cells ◽  
Tether Extraction ◽  
Surface Receptor ◽  
Microvascular Endothelial ◽  
Pulling Force

Membrane tethers were found to be extracted from leukocytes and macrovascular endothelial cells (e.g., human umbilical vein endothelial cells or HUVECs) when a point pulling force was exerted. These tethers stabilize leukocyte rolling on the endothelium during the inflammatory response. However, little is known about tether extraction from other vascular cells like microvascular endothelial cells (MECs). In this study, we extracted tethers from both adult and neonatal dermal MECs with the micropipette aspiration technique. We found a linear relationship between the pulling force and tether growth velocity for both cell lines. This constitutive relationship is mainly determined by the membrane mechanical property and the underlying actin-based cytoskeleton for both attached and suspended endothelial cells. It is independent of cell surface receptor type, attachment state, cytokine stimulation, or cell lineage. For both types of MECs, the threshold forces are ∼50 pN and the effective viscosities are around 0.5 pN·s/μm. These results, which are close to what was obtained from HUVECs, indicate that homogeneity is preserved in terms of tether extraction among different types of endothelial cells, and simultaneous tethers are likely extracted when leukocytes roll on either microvascular or macrovascular surfaces.

scholarly journals Streptococcus suis Serotype 2 Interactions with Human Brain Microvascular Endothelial Cells

2000 ◽  
Vol 68 (2) ◽  
pp. 637-643 ◽  
Author(s):  
Nathalie Charland ◽  
Victor Nizet ◽  
Craig E. Rubens ◽  
Kwang Sik Kim ◽  
Sonia Lacouture ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Damage ◽  
Umbilical Vein ◽  
Bacterial Species ◽  
Streptococcus Suis ◽  
Neonatal Meningitis ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Group B Streptococci ◽  
Microvascular Endothelial

ABSTRACT Streptococcus suis serotype 2 is a worldwide causative agent of many forms of swine infection and is also recognized as a zoonotic agent causing human disease, including meningitis. The pathogenesis of S. suis infections is poorly understood. Bacteria circulate in the bloodstream in the nonimmune host until they come in contact with brain microvascular endothelial cells (BMEC) forming the blood-brain barrier. The bacterial polysaccharide capsule confers antiphagocytic properties. It is known that group B streptococci (GBS) invade and damage BMEC, which may be a primary step in the pathogenesis of neonatal meningitis. Interactions betweenS. suis and human endothelial cells were studied to determine if they differ from those between GBS and endothelial cells. Invasion assays performed with BMEC and human umbilical vein endothelial cells demonstrated that unlike GBS, S. suisserotype 2 could not invade either type of cell. Adherence assays showed that S. suis adhered only to BMEC, whereas GBS adhered to both types of cell. These interactions were not affected by the presence of a capsule, since acapsular mutants from both bacterial species adhered similarly compared to the wild-type strains. Lactate dehydrogenase release measurements indicated that some S. suis strains were highly cytotoxic for BMEC, even more than GBS, whereas others were not toxic at all. Cell damage was related to suilysin (S. suis hemolysin) production, since only suilysin-producing strains were cytotoxic and cytotoxicity could be inhibited by cholesterol and antisuilysin antibodies. It is possible that hemolysin-positive S. suis strains use adherence and suilysin-induced BMEC injury, as opposed to direct cellular invasion, to proceed from the circulation to the central nervous system.

CULTURED HUMAN MICROVASCULAR ENDOTHELIAL CELLS SYNTHESIZE CYCLOOXYGENASE METABOLITES FROM ARACHIDONIC ACID IN RESPONSE TO LEUKOTRIENES C4 AND D4

1987 ◽  
Author(s):  
L O Carreras ◽  
J Maclouf ◽  
G Tobelem ◽  
J P Caen
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Time Course ◽  
Umbilical Vein ◽  
Calcium Ionophore ◽  
Microvascular Endothelial Cells ◽  
Ionophore A23187 ◽  
Total Production ◽  
Dependent Manner ◽  
Microvascular Endothelial

Several investigators have demonstrated that endothelial cells have heterogeneous intrinsic properties depending on their vascular origin. In this respect, very limited knowledge exists concerning the production of eicosanoids by human microvascular endothelial cells (HMEC). The aim of this study was to determine: 1) the pattern of the production of cyclooxygenase metabolites by cultured HMEC from omental adipose tissue as compared to the classical study of human umbilical vein endothelial cells (HUVEC); 2) the modification of this metabolism upon leukotrienes (LTs) stimulation. Cultured HMEC produced prostaglandin (PG) E2, PGF2 , 6-keto-PGF1 , and PGD2 (measured by enzymoimmunoassay). In basal conditions, PGD2 was the main product released in the supernatant. Upon stimulation with thrombin, arachidonic acid and calcium ionophore A23187, a marked increase in the production of PGE2, PGF2 , and 6-keto-PGFj , was observed; these results were quite different from HUVEC. In contrast, PGD2 remained unchanged under our experimental conditions and thromboxane B2 was always undetectable. In all cases, the release of PGE2 and PGF2 , was higher than that of 6-keto-PGFj . A considerable amount of the metabolites produced remained cell-associated. The total production (release + cell bound) of cyclooxygenase products was stimulated by LTC4 and LTD4 in a dose-dependent manner (10-9 to 10-6 M). The production of PGD2 was unchanged. LTC4 and LTD4 were almost equally potent, but LTB4 was unable to stimulate PG synthesis (n=4). The production of metabolites induced by 1 uM LTC4 or LTD4 was even higher than that obtained in the presence of high concentrations of thrombin (5 U/ml). This contrasted with the more pronounced stimulation of thrombin on HUVEC as compared to LTs. In the kinetic studies (n=2) we have observed a slow time-course of release of PGE2 and 6-keto-PGF1 into the supernatant of LTs-stimulated HMEC (half-maximal formation at 14-15 min). The stimulatory activity of LTC4 and LTD4 on the production of vasoactive cyclooxygenase metabolites by HMEC could be relevant in inflammatory processes.

scholarly journals Thrombin Modulates the Expression of a Set of Genes Including Thrombospondin-1 in Human Microvascular Endothelial Cells

2005 ◽  
Vol 280 (23) ◽  
pp. 22172-22180 ◽  
Author(s):  
Joseph N. McLaughlin ◽  
Maria R. Mazzoni ◽  
John H. Cleator ◽  
Laurie Earls ◽  
Ana Luisa Perdigoto ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Umbilical Vein ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelium ◽  
Disease States ◽  
Similar Expression Profile ◽  
Thrombospondin 1 ◽  
Microvascular Endothelial

Thrombospondin-1 (THBS1) is a large extracellular matrix glycoprotein that affects vasculature systems such as platelet activation, angiogenesis, and wound healing. Increases in THBS1 expression have been liked to disease states including tumor progression, atherosclerosis, and arthritis. The present study focuses on the effects of thrombin activation of the G-protein-coupled, protease-activated receptor-1 (PAR-1) on THBS1 gene expression in the microvascular endothelium. Thrombin-induced changes in gene expression were characterized by microarray analysis of ∼11,000 different human genes in human microvascular endothelial cells (HMEC-1). Thrombin induced the expression of a set of at least 65 genes including THBS1. Changes in THBS1 mRNA correlated with an increase in the extracellular THBS1 protein concentration. The PAR-1-specific agonist peptide (TFLLRNK-PDK) mimicked thrombin stimulation of THBS1 expression, suggesting that thrombin signaling is through PAR-1. Further studies showed THBS1 expression was sensitive to pertussis toxin and protein kinase C inhibition indicating Gi/o- and Gq-mediated pathways. THBS1 up-regulation was also confirmed in human umbilical vein endothelial cells stimulated with thrombin. Analysis of the promoter region of THBS1 and other genes of similar expression profile identified from the microarray predicted an EBOX/EGRF transcription model. Expression of members of each family, MYC and EGR1, respectively, correlated with THBS1 expression. These results suggest thrombin formed at sites of vascular injury increases THBS1 expression into the extracellular matrix via activation of a PAR-1, Gi/o, Gq, EBOX/EGRF-signaling cascade, elucidating regulatory points that may play a role in increased THBS1 expression in disease states.

scholarly journals Human umbilical vein and dermal microvascular endothelial cells show heterogeneity in response to PKC activation

1997 ◽  
Vol 273 (4) ◽  
pp. C1233-C1240 ◽  
Author(s):  
Justin C. Mason ◽  
Helen Yarwood ◽  
Katharine Sugars ◽  
Dorian O. Haskard
Keyword(s):  
Endothelial Cells ◽  
Adhesion Molecule ◽  
Morphological Changes ◽  
Umbilical Vein ◽  
Intercellular Adhesion Molecule ◽  
Microvascular Endothelial Cells ◽  
Intercellular Adhesion Molecule 1 ◽  
Human Umbilical Vein ◽  
Microvascular Endothelial ◽  
Pkc Activation

Changes in endothelial cell (EC) phenotype are central to the function of endothelium in inflammation. Although these events mainly occur in the microvasculature, previous studies have predominantly used large-vessel EC. Using enzyme-linked immunosorbent and flow cytometric assays, we compared the responses of human umbilical vein endothelial cells (HUVEC) and dermal microvascular endothelial cells (DMEC) to the activation of protein kinase C (PKC). Stimulation with phorbol 12,13-dibutyrate and more selective PKC agonists, including 12-deoxyphorbol 13-phenylacetate 20-acetate (dPPA), induced morphological changes and proliferation in both EC types. PKC activation induced a marked increase in Thy-1 expression on DMEC and only a moderate rise on HUVEC. Furthermore, heterogeneity in the induction of the adhesion molecules intercellular adhesion molecule 1, vascular cell adhesion molecule 1 (VCAM-1), and E-selectin between the two EC types following activation of PKC was demonstrated. In particular, E-selectin and VCAM-1 were significantly upregulated on HUVEC but not DMEC. The data indicate that the PKC pathway is unlikely to be important for E-selectin and VCAM-1 expression in the microvasculature but are consistent with a role for PKC in angiogenesis. This diversity in signaling in response to PKC activation may depend on differential utilization of PKC isozymes and may facilitate specialized endothelial responses.

Clot Lysis Mediated by Cultured Human Microvascular Endothelial Cells

1988 ◽  
Vol 60 (03) ◽  
pp. 463-467 ◽  
Author(s):  
Wolfgang Speiser ◽  
Elisabeth Anders ◽  
Bernd R Binder ◽  
Gert Müller-Berghaus
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Shape Change ◽  
Umbilical Vein ◽  
Cell Types ◽  
Microvascular Endothelial Cells ◽  
Clot Lysis ◽  
Microvascular Endothelial ◽  
Umbilical Vein Endothelial Cells ◽  
Fibrin Clots

SummaryThe lysis of fibrin clots on the surface of cultured human omental tissue microvascular endothelial cells (HOTMEC) and cultured human umbilical vein endothelial cells (HUVEC) was studied. Fibrin clots were made by mixing fibrinogen, plasminogen and thrombin on the surface of both cell types. Clot lysis was seen only on the surface of HOTMEC, which were found to synthesize about 100-fold more tissue plasminogen activator (tPA) antigen than HUVEC. Clot lysis of HOTMEC could be blocked by anti-tPA IgG but was not affected by the incorporation of exogenous plasminogen activator (PAI) into the clot in concentrations (75 arbitrary units) exceeding the tPA activity (21 ± 2.5 IU) of the cells. Thus, it is likely that tPA secreted by HOTMEC is protected from inhibition by PAI in the presence of fibrin and endothelial cells. The stimulation of EC to release an excess of tPA over PAI, in contrast to the secretion of an excess of PAI over tPA found in unstimulated cells in the absence of fibrin, is obviously no prerequisite for the initiation of fibrinolysis on the surface of HOTMEC. As thrombin was used for clot formation, its influence on tPA and PAI synthesis of both cell types was investigated. In contrast to HOTMEC, which were not affected by Α-thrombin, HUVEC revealed a dose-dependent increase in tPA and PAI synthesis upon incubation with the enzyme. This increase in tPA production by HUVEC was not sufficient to lyse the clots within 48 hours. Furthermore, HUVEC. behaved differently towards thrombin as these cells in contrast to HOTMEC revealed the typical shape change reaction upon incubation with the enzyme

Sulfated glycoconjugates enhance CD36-dependent adhesion ofPlasmodium falciparum–infected erythrocytes to human microvascular endothelial cells

Blood ◽  
2000 ◽  
Vol 96 (1) ◽  
pp. 327-333 ◽  
Author(s):  
Christopher John McCormick ◽  
Christopher I. Newbold ◽  
Anthony R. Berendt
Keyword(s):  
Monoclonal Antibody ◽  
Endothelial Cells ◽  
Dextran Sulfate ◽  
Umbilical Vein ◽  
Microvascular Endothelial Cells ◽  
Cos Cells ◽  
Ofplasmodium Falciparum ◽  
Microvascular Endothelial ◽  
Umbilical Vein Endothelial Cells ◽  
Negative Cell Line

A novel adhesive pathway that enhances the adhesion ofPlasmodium falciparum-infected erythrocytes (IEs) to endothelial cells has been identified. The sulfated glycoconjugates heparin, fucoidan, dextran sulfate 5000, and dextran sulfate 500 000 caused a dramatic increase in adhesion of IEs to human dermal microvascular endothelial cells. The same sulfated glycoconjugates had little effect on IE adhesion to human umbilical vein endothelial cells, a CD36-negative cell line. The effect was abolished by a monoclonal antibody directed against CD36, suggesting that enhanced adhesion to endothelium is dependent on CD36. No effect was observed on adhesion to purified platelet CD36 cells immobilized on plastic. The same sulfated glycoconjugates enhanced adhesion of infected erythrocytes to COS cells transfected with CD36, and this was inhibited by the CD36 monoclonal antibody. These findings demonstrate a role for sulfated glycoconjugates in endothelial adherence that may be important in determining the location and magnitude of sequestration through endogenous carbohydrates. In addition, they highlight possible difficulties that may be encountered from the proposed use of sulfated glycoconjugates as antiadhesive agents in patients with severe malaria.

scholarly journals CD36 Mediates the In Vitro Inhibitory Effects of Thrombospondin-1 on Endothelial Cells

1997 ◽  
Vol 138 (3) ◽  
pp. 707-717 ◽  
Author(s):  
David W. Dawson ◽  
S. Frieda A. Pearce ◽  
Ruiqin Zhong ◽  
Roy L. Silverstein ◽  
William A. Frazier ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fusion Proteins ◽  
Solid Phase ◽  
Umbilical Vein ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Microvascular Endothelial Cells ◽  
Thrombospondin 1 ◽  
Microvascular Endothelial

Thrombospondin-1 (TSP-1) is a naturally occurring inhibitor of angiogenesis that is able to make normal endothelial cells unresponsive to a wide variety of inducers. Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells. Both IgG antibodies against CD36 and glutathione-S-transferase–CD36 fusion proteins that contain the TSP-1 binding site blocked the ability of intact TSP-1 and its active peptides to inhibit the migration of cultured microvascular endothelial cells. In addition, antiangiogenic TSP-1 peptides inhibited the binding of native TSP-1 to solid phase CD36 and its fusion proteins, as well as to CD36-expressing cells. Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM∅, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells. Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation. This work demonstrates that endothelial CD36, previously thought to be involved only in adhesion and scavenging activities, may be essential for the inhibition of angiogenesis by thrombospondin-1.

Sulfated glycoconjugates enhance CD36-dependent adhesion ofPlasmodium falciparum–infected erythrocytes to human microvascular endothelial cells

Blood ◽  
2000 ◽  
Vol 96 (1) ◽  
pp. 327-333 ◽  
Author(s):  
Christopher John McCormick ◽  
Christopher I. Newbold ◽  
Anthony R. Berendt
Keyword(s):  
Monoclonal Antibody ◽  
Endothelial Cells ◽  
Dextran Sulfate ◽  
Umbilical Vein ◽  
Microvascular Endothelial Cells ◽  
Cos Cells ◽  
Ofplasmodium Falciparum ◽  
Microvascular Endothelial ◽  
Umbilical Vein Endothelial Cells ◽  
Negative Cell Line

Abstract A novel adhesive pathway that enhances the adhesion ofPlasmodium falciparum-infected erythrocytes (IEs) to endothelial cells has been identified. The sulfated glycoconjugates heparin, fucoidan, dextran sulfate 5000, and dextran sulfate 500 000 caused a dramatic increase in adhesion of IEs to human dermal microvascular endothelial cells. The same sulfated glycoconjugates had little effect on IE adhesion to human umbilical vein endothelial cells, a CD36-negative cell line. The effect was abolished by a monoclonal antibody directed against CD36, suggesting that enhanced adhesion to endothelium is dependent on CD36. No effect was observed on adhesion to purified platelet CD36 cells immobilized on plastic. The same sulfated glycoconjugates enhanced adhesion of infected erythrocytes to COS cells transfected with CD36, and this was inhibited by the CD36 monoclonal antibody. These findings demonstrate a role for sulfated glycoconjugates in endothelial adherence that may be important in determining the location and magnitude of sequestration through endogenous carbohydrates. In addition, they highlight possible difficulties that may be encountered from the proposed use of sulfated glycoconjugates as antiadhesive agents in patients with severe malaria.

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