Overexpression of interleukin‐10 in engineered macrophages protects endothelial cells against LPS‐induced injury in vitro

Endothelial cells (ECs) are believed to be an important component in the protection from lipopolysaccharide (LPS)-induced endotoxic shock. However, the cellular and molecular mechanism is not well defined. Here, we report that signal transducer and activator of transcription (STAT) 3 is an essential regulator of the antiinflammatory function of ECs in systemic immunity. Because STAT3 deficiency results in early embryonic lethality, we have generated mice with a conditional STAT3 deletion in endothelium (STAT3E−/−). STAT3E−/− mice are healthy and fertile, and isolated ECs initiate normal tube formation in vitro. Conditional endothelial but not organ-specific (i.e., hepatocyte or cardiomyocyte) STAT3 knockout mice show an increased susceptibility to lethality after LPS challenge. The LPS response in STAT3E−/− mice shows exaggerated inflammation and leukocyte infiltration in multiple organs combined with elevated activity of serum alanine aminotransferase and aspartate aminotransferase, indicating organ damage. Concomitantly, proinflammatory cytokines are produced at an exaggerated level and for a prolonged period. This defect cannot be explained by lack of antiinflammatory cytokines, such as interleukin 10 and transforming growth factor β. Instead, we have shown that a soluble activity derived from endothelia and dependent on STAT3 is critical for suppression of interferon γ. These data define STAT3 signaling within endothelia as a critical antiinflammatory mediator and provide new insight to the protective function of ECs in inflammation.

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Activation of Endothelium by Borrelia burgdorferi In Vitro Enhances Transmigration of Specific Subsets of T Lymphocytes

Infection and Immunity ◽

10.1128/iai.69.4.2190-2197.2001 ◽

2001 ◽

Vol 69 (4) ◽

pp. 2190-2197 ◽

Cited By ~ 14

Author(s):

Edna I. Gergel ◽

Martha B. Furie

Keyword(s):

Endothelial Cells ◽

T Lymphocytes ◽

Borrelia Burgdorferi ◽

Blood Vessel ◽

Umbilical Vein ◽

Active Role ◽

Interleukin 10 ◽

Fourfold Increase ◽

Umbilical Vein Endothelial Cells

ABSTRACT Lyme disease, caused by Borrelia burgdorferi, is characterized by the accumulation of lymphocytes and monocytes in the affected tissue. Endothelial cells line the blood vessel walls and control the trafficking of inflammatory leukocytes from the blood into the surrounding tissues. A model of the blood vessel wall, consisting of human umbilical vein endothelial cells (HUVEC) grown on amniotic connective tissue, was utilized to examine the effects of B. burgdorferi on the transendothelial migration of T lymphocytes. Maximal migration occurred when the HUVEC-amnion cultures were preincubated with B. burgdorferi for 24 h and T lymphocytes were added for an additional 4 h, yielding a two- to fourfold increase compared to migration across unstimulated cultures. The number of T lymphocytes that migrated was proportional to the number added. The anti-inflammatory cytokine interleukin 10 (IL-10), added during activation of the HUVEC, significantly diminished (by an average of 70% ± 21%) the migration of T lymphocytes across endothelium stimulated for 8 or 24 h with B. burgdorferi, but not IL-1. Compared to the initially added population of T lymphocytes, the population that migrated across untreated endothelium or HUVEC activated with B. burgdorferi or IL-1 contained a significantly smaller percentage of CD45RA+RO− (naı̈ve) cells and a greater proportion of CD45RA+RO+ cells. The migratory population was also enriched for CD8+ T lymphocytes when the endothelium was incubated with either control medium or B. burgdorferi, but not IL-1. B. burgdorferi thus activates endothelium in a manner that promotes the transmigration of T lymphocytes, and IL-10 inhibits this activation. These data further suggest that endothelium plays an active role in promoting the recruitment of specific subpopulations of T lymphocytes.

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IN VITRO ACTIVITY OF BIOACTIVE MOLECULES INCORPORATED INTO POLY (3-HYDROXYBUTYRATE-CO-3-HYDROXYVALERATE)/ POLY(ε-CAPROLACTONE) SCAFFOLDS

Complex Issues of Cardiovascular Diseases ◽

10.17802/2306-1278-2018-7-2-89-101 ◽

2018 ◽

Vol 7 (2) ◽

pp. 89-101 ◽

Cited By ~ 1

Author(s):

L. V. Antonova ◽

V. G. Matveeva ◽

E. A. Velikanova ◽

M. Y. Khanova ◽

V. V. Sevostyanova ◽

...

Keyword(s):

Endothelial Cells ◽

Growth Factor ◽

Cellular Response ◽

Intercellular Junctions ◽

Interleukin 10 ◽

Bioactive Molecules ◽

Vitro Activity ◽

Superior Performance ◽

In Vitro Activity

Background We fabricated biodegradable, bioactive scaffolds to guide the differentiation of endothelial progenitor cells. Aim To study in vitro activity of the bioactive factors incorporated into the poly (3-hydroxubutyrate-co-3-hydroxyvalerate)/poly(ε-caprolactone) (PHBV/PCL) scaffolds. Methods Nonwoven scaffolds were blended of PHBV and PCL utilizing either separate or combined incorporation of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and stromal cell-derived factor-1α (SDF-1α) by emulsion electrospinning. We further studied adhesion, viability, and proliferation of EA.hy 926 endothelial cells cultured on these scaffolds and evaluated vasculogenesis, cell index, and secretory profile in response to the addition of abovementioned bioactive factors. Results We showed that VEGF, bFGF, and SDF-1α retain their bioactivity upon the incorporation into the PHBV/PCL scaffolds. Scaffolds with all three bioactive factors incorporated demonstrated superior performance in comparison with those containing any of these factors alone. Diffusion of the bioactive factors into the culture medium stimulated the secretion of interleukin-10, and VE-cadherin by endothelial cells that indicated anti-inflammatory response and tight intercellular junctions. We also detected the low level of secreted VEGF-A from the scaffolds with VEGF suggestive of its physiological regulation. Conclusion Bioactive factors retain their bioactivity upon the incorporation into the PHBV/ PCL scaffolds. Combination of VEGF, bFGF, and SDF-1a improves cellular response compared to the incorporation of any of these factors alone.

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Human Cytomegalovirus Interleukin-10 Downregulates Metalloproteinase Activity and Impairs Endothelial Cell Migration and Placental Cytotrophoblast Invasiveness In Vitro

Journal of Virology ◽

10.1128/jvi.78.6.2831-2840.2004 ◽

2004 ◽

Vol 78 (6) ◽

pp. 2831-2840 ◽

Cited By ~ 91

Author(s):

Takako Yamamoto-Tabata ◽

Susan McDonagh ◽

Hsin-Ti Chang ◽

Susan Fisher ◽

Lenore Pereira

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Human Cytomegalovirus ◽

Laboratory Strain ◽

Interleukin 10 ◽

Endothelial Cell Migration ◽

Microvascular Endothelial Cells ◽

Placental Development ◽

Microvascular Endothelial

ABSTRACT At the uterine-placental interface, fetal cytotrophoblasts invade the decidua, breach maternal blood vessels, and form heterotypic contacts with uterine microvascular endothelial cells. In early gestation, differentiating- invading cytotrophoblasts produce high levels of matrix metalloproteinase 9 (MMP-9), which degrades the extracellular matrix and increases the invasion depth. By midgestation, when invasion is complete, MMP levels are reduced. Cytotrophoblasts also produce human interleukin-10 (hIL-10), a pleiotropic cytokine that modulates immune responses, helping to protect the fetal hemiallograft from rejection. Human cytomegalovirus (CMV) is often detected at the uterine-placental interface. CMV infection impairs cytotrophoblast differentiation and invasion, altering the expression of the cell adhesion and immune molecules. Here we report that infection with a clinical CMV strain, VR1814, but not a laboratory strain, AD169, downregulates MMP activity in uterine microvascular endothelial cells and differentiating-invading cytotrophoblasts. Infected cytotrophoblasts expressed CMV IL-10 (cmvIL-10) mRNA and secreted the viral cytokine, which upregulated hIL-10. Functional analyses showed that cmvIL-10 treatment impaired migration in endothelial cell wounding assays and cytotrophoblast invasion of Matrigel in vitro. Comparable changes occurred in cells that were exposed to recombinant hIL-10 or cmvIL-10. Our results show that cmvIL-10 decreases MMP activity and dysregulates the cell-cell and/or cell-matrix interactions of infected cytotrophoblasts and endothelial cells. Reduced MMP activity early in placental development could impair cytotrophoblast remodeling of the uterine vasculature and eventually restrict fetal growth in affected pregnancies.

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Ultrastructural studies on the interaction of haemophilus influenzae with human endothelial cells in vitro

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010016073x ◽

1990 ◽

Vol 48 (3) ◽

pp. 636-637

Author(s):

D.J.P. Ferguson ◽

M. Virji ◽

H. Kayhty ◽

E.R. Moxon

Keyword(s):

Endothelial Cells ◽

Haemophilus Influenzae ◽

Intercellular Junctions ◽

Blood Stream ◽

Ultrastructural Studies ◽

Endothelial Barriers ◽

Serotype B ◽

Meningitis In Children ◽

Respiratory Epithelial

Haemophilus influenzae is a human pathogen which causes meningitis in children. Systemic H. influenzae infection is largely confined to encapsulated serotype b organisms and is a major cause of meningitis in the U.K. and elsewhere. However, the pathogenesis of the disease is still poorly understood. Studies in the infant rat model, in which intranasal challenge results in bacteraemia, have shown that H. influenzae enters submucosal tissues and disseminates to the blood stream within minutes. The rapidity of these events suggests that H. influenzae penetrates both respiratory epithelial and endothelial barriers with great efficiency. It is not known whether the bacteria penetrate via the intercellular junctions, are translocated within the cells or carried across the cellular barrier in 'trojan horse' fashion within phagocytes. In the present studies, we have challenged cultured human umbilical cord_vein endothelial cells (HUVECs) with both capsulated (b+) and capsule-deficient (b-) isogenic variants of one strain of H. influenzae in order to investigate the interaction between the bacteria and HUVEC and the effect of the capsule.

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Anti-metalloproteinase-9 Activities of Selected Indonesian Zingiberaceae Rhizome Extracts in Lipopolysaccharide-induced Human Vascular Endothelial Cells In Vitro

Planta Medica ◽

10.1055/s-0031-1282610 ◽

2011 ◽

Vol 77 (12) ◽

Author(s):

Y Yanti ◽

N Steven ◽

A Wiharja ◽

S Fajarianto

Keyword(s):

Endothelial Cells ◽

Vascular Endothelial Cells ◽

Vascular Endothelial ◽

Metalloproteinase 9

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Adverse Influence of Cigarette Smoking on the Endothelium

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649654 ◽

1993 ◽

Vol 70 (04) ◽

pp. 707-711 ◽

Cited By ~ 77

Author(s):

Andrew D Blann ◽

Charles N McCollum

Keyword(s):

Endothelial Cells ◽

Von Willebrand Factor ◽

Tobacco Smoke ◽

Lipid Peroxides ◽

Short Exposure ◽

Acute Effects ◽

Adverse Influence ◽

Von Willebrand ◽

Willebrand Factor

SummaryThe effect of smoking on the blood vessel intima was examined by comparing indices of endothelial activity in serum from smokers with that from non-smokers. Serum from smokers contained higher levels of von Willebrand factor (p <0.01), the smoking markers cotinine (p <0.02) and thiocyanate (p <0.01), and was more cytotoxic to endothelial cells in vitro (p <0.02) than serum from non-smokers. The acute effects of smoking two unfiltered medium tar cigarettes was to briefly increase von Willebrand factor (p <0.001) and cytotoxicity of serum to endothelial cells in vitro (p <0.005), but lipid peroxides or thiocyanate were not increased by this short exposure to tobacco smoke. Although there were correlations between von Willebrand factor and smokers consumption of cigarettes (r = 0.28, p <0.02), number of years smoking (r = 0.41, p <0.001) and cotinine (r = 0.45, p <0.01), the tissue culture of endothelial cells with physiological levels of thiocyanate or nicotine suggested that these two smoking markers were not cytotoxic. They are therefore unlikely to be directly responsible for increased von Willebrand factor in the serum of smokers. We suggest that smoking exerts a deleterious influence on the endothelium and that the mechanism is complex.

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Perturbation of Platelet Adhesion to Endothelial Cells by Plasminogen Activation In Vitro

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657655 ◽

1997 ◽

Vol 78 (02) ◽

pp. 934-938 ◽

Cited By ~ 5

Author(s):

Hsiun-ing Chen ◽

Yueh-I Wu ◽

Yu-Lun Hsieh ◽

Guey-Yueh Shi ◽

Meei-Jyh Jiang ◽

...

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Platelet Adhesion ◽

Treatment Time ◽

Shape Change ◽

Umbilical Vein ◽

Tissue Type ◽

Apical Surface ◽

Plasminogen Activation

SummaryTo investigate whether the endothelium-platelet interactions may be altered by plasminogen activation, cultured human umbilical vein endothelial cells (ECs) were treated with tissue-type plasminogen activator (t-PA) in the presence of plasminogen, and platelet adhesion to ECs was subsequently measured by using a tapered flow chamber. Our results demonstrated that platelets adhered more readily to t-PA treated EC monolayer than to the control monolayer at all shear stress levels tested. This phenomenon was treatment time-dependent and dose-dependent, and it could be blocked by adding plasmin inhibitors, such as e-amino caproic acid and aprotinin. Adherent platelets on t-PA treated EC monolayer underwent more severe shape change than those on the control monolayer. While the extracellular matrix directly treated with t-PA attracted less platelets than the control matrix did, platelet adhesion to the matrix that was produced by t-PA-treated ECs was unaltered. These data suggest that t-PA treatment on ECs compromised antiplatelet-adhesion capability on their apical surface without altering the reactivity of their extracellular matrix towards platelets.

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Desmopressin (DDAVP) Enhances Platelet Adhesion to the Extracellular Matrix of Cultured Human Endothelial Cells through Increased Expression of Tissue Factor

Thrombosis and Haemostasis ◽

10.1055/s-0038-1656088 ◽

1997 ◽

Vol 77 (05) ◽

pp. 0975-0980 ◽

Cited By ~ 23

Author(s):

Angel Gálvez ◽

Goretti Gómez-Ortiz ◽

Maribel Díaz-Ricart ◽

Ginés Escolar ◽

Rogelio González-Sarmiento ◽

...

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Tissue Factor ◽

Platelet Adhesion ◽

Umbilical Vein ◽

Procoagulant Activity ◽

Experimental Conditions ◽

Chromogenic Assay

SummaryThe effect of desmopressin (DDAVP) on thrombogenicity, expression of tissue factor and procoagulant activity (PCA) of extracellular matrix (ECM) generated by human umbilical vein endothelial cells cultures (HUVEC), was studied under different experimental conditions. HUVEC were incubated with DDAVP (1, 5 and 30 ng/ml) and then detached from their ECM. The reactivity towards platelets of this ECM was tested in a perfusion system. Coverslips covered with DD A VP-treated ECMs were inserted in a parallel-plate chamber and exposed to normal blood anticoagulated with low molecular weight heparin (Fragmin®, 20 U/ml). Perfusions were run for 5 min at a shear rate of 800 s1. Deposition of platelets on ECMs was significantly increased with respect to control ECMs when DDAVP was used at 5 and 30 ng/ml (p <0.05 and p <0.01 respectively). The increase in platelet deposition was prevented by incubation of ECMs with an antibody against human tissue factor prior to perfusion. Immunofluorescence studies positively detected tissue factor antigen on DDAVP derived ECMs. A chromogenic assay performed under standardized conditions revealed a statistically significant increase in the procoagulant activity of the ECMs produced by ECs incubated with 30 ng/ml DDAVP (p <0.01 vs. control samples). Northern blot analysis revealed increased levels of tissue factor mRNA in extracts from ECs exposed to DDAVP. Our data indicate that DDAVP in vitro enhances platelet adhesion to the ECMs through increased expression of tissue factor. A similar increase in the expression of tissue factor might contribute to the in vivo hemostatic effect of DDAVP.

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Effects of Dipyrone on Prostaglandin Production by Human Platelets and Cultured Bovine Aortic Endothelial Cells

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657338 ◽

1983 ◽

Vol 49 (02) ◽

pp. 132-137 ◽

Cited By ~ 16

Author(s):

A Eldor ◽

G Polliack ◽

I Vlodavsky ◽

M Levy

Keyword(s):

Endothelial Cells ◽

Arachidonic Acid ◽

Competitive Inhibition ◽

Inhibitory Effect ◽

Human Platelets ◽

Ionophore A23187 ◽

Aortic Endothelial Cells ◽

Bovine Aortic Endothelial Cells ◽

Aortic Endothelial

SummaryDipyrone and its metabolites 4-methylaminoantipyrine, 4-aminoantipyrine, 4-acetylaminoantipyrine and 4-formylaminoan- tipyrine inhibited the formation of thromboxane A2 (TXA2) during in vitro platelet aggregation induced by ADP, epinephrine, collagen, ionophore A23187 and arachidonic acid. Inhibition occurred after a short incubation (30–40 sec) and depended on the concentration of the drug or its metabolites and the aggregating agents. The minimal inhibitory concentration of dipyrone needed to completely block aggregation varied between individual donors, and related directly to the inherent capacity of their platelets to synthesize TXA2.Incubation of dipyrone with cultured bovine aortic endothelial cells resulted in a time and dose dependent inhibition of the release of prostacyclin (PGI2) into the culture medium. However, inhibition was abolished when the drug was removed from the culture, or when the cells were stimulated to produce PGI2 with either arachidonic acid or ionophore A23187.These results indicate that dipyrone exerts its inhibitory effect on prostaglandins synthesis by platelets or endothelial cells through a competitive inhibition of the cyclooxygenase system.

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