Microparticle-mediated VZV propagation and endothelial activation

ObjectiveVaricella zoster virus (VZV) can spread anterogradely and infect cerebral arteries causing VZV vasculopathy and arterial ischemic stroke. In this study, we tested the hypothesis that virus-infected cerebrovascular fibroblasts undergo phenotypic changes that promote vascular remodeling and facilitate virus transmission in an in vitro model of VZV vasculopathy. The aims of this project were therefore to examine the changes that virus-infected human brain adventitial vascular fibroblasts (HBVAFs) undergo in an in vitro model of VZV vasculopathy and to identify disease biomarkers relating to VZV-related vasculopathy.MethodsHBVAFs were infected with VZV, and their ability to migrate, proliferate, transdifferentiate, and interact with endothelial cells was studied with flow cytometry. Microparticles (MPs) released from these cells were isolated and imaged with transmission electron microscopy, and their protein content was analyzed with mass spectrometry. Circulating MP profiles were also studied in children with VZV and non-VZV vasculopathy and compared with controls.ResultsVZV-infected HBVAFs transdifferentiated into myofibroblasts with enhanced proliferative and migratory capacity. Interaction of VZV-infected HBVAFs with endothelial cells resulted in endothelial dysfunction. These effects were, in part, mediated by the release of MPs from VZV-infected HBVAFs. These MPs contained VZV virions that could transmit VZV to neighboring cells, highlighting a novel model of VZV cell-to-cell viral dissemination. MPs positive for VZV were significantly higher in children with VZV-related vasculopathy compared to children with non-VZV vasculopathy (p = 0.01) and controls (p = 0.007).ConclusionsVZV-infected HBVAFs promote vascular remodeling and facilitate virus transmission. These effects were mediated by the release of apoptotic MPs that could transmit VZV infection to neighboring cells through a Trojan horse means of productive viral infection. VZV+ MPs may represent a disease biomarker worthy of further study.

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Generation of a Novel In Vitro Model to Study Endothelial Dysfunction from Atherothrombotic Specimens

Cardiovascular Drugs and Therapy ◽

10.1007/s10557-021-07151-9 ◽

2021 ◽

Author(s):

Susan Gallogly ◽

Takeshi Fujisawa ◽

John D. Hung ◽

Mairi Brittan ◽

Elizabeth M. Skinner ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Dysfunction ◽

In Vitro Model ◽

Umbilical Vein ◽

Coronary Syndrome ◽

Coronary Endothelial Cells ◽

Vitro Model ◽

Umbilical Vein Endothelial Cells

Abstract Purpose Endothelial dysfunction is central to the pathogenesis of acute coronary syndrome. The study of diseased endothelium is very challenging due to inherent difficulties in isolating endothelial cells from the coronary vascular bed. We sought to isolate and characterise coronary endothelial cells from patients undergoing thrombectomy for myocardial infarction to develop a patient-specific in vitro model of endothelial dysfunction. Methods In a prospective cohort study, 49 patients underwent percutaneous coronary intervention with thrombus aspiration. Specimens were cultured, and coronary endothelial outgrowth (CEO) cells were isolated. CEO cells, endothelial cells isolated from peripheral blood, explanted coronary arteries, and umbilical veins were phenotyped and assessed functionally in vitro and in vivo. Results CEO cells were obtained from 27/37 (73%) atherothrombotic specimens and gave rise to cells with cobblestone morphology expressing CD146 (94 ± 6%), CD31 (87 ± 14%), and von Willebrand factor (100 ± 1%). Proliferation of CEO cells was impaired compared to both coronary artery and umbilical vein endothelial cells (population doubling time, 2.5 ± 1.0 versus 1.6 ± 0.3 and 1.2 ± 0.3 days, respectively). Cell migration was also reduced compared to umbilical vein endothelial cells (29 ± 20% versus 85±19%). Importantly, unlike control endothelial cells, dysfunctional CEO cells did not incorporate into new vessels or promote angiogenesis in vivo. Conclusions CEO cells can be reliably isolated and cultured from thrombectomy specimens in patients with acute coronary syndrome. Compared to controls, patient-derived coronary endothelial cells had impaired capacity to proliferate, migrate, and contribute to angiogenesis. CEO cells could be used to identify novel therapeutic targets to enhance endothelial function and prevent acute coronary syndromes.

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Minocycline induces protective autophagy in vascular endothelial cells exposed to an in vitro model of ischemia/reperfusion-induced injury

Biomedical Reports ◽

10.3892/br.2015.554 ◽

2015 ◽

Vol 4 (2) ◽

pp. 173-177 ◽

Cited By ~ 13

Author(s):

WENBIN DONG ◽

SHIGENG XIAO ◽

MIN CHENG ◽

XIAODI YE ◽

GAOLI ZHENG

Keyword(s):

Endothelial Cells ◽

In Vitro Model ◽

Vascular Endothelial Cells ◽

Ischemia Reperfusion ◽

Vascular Endothelial ◽

Vitro Model

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Passage of low density lipoproteins through monolayers of human arterial endothelial cells. Effects of vasoactive substances in an in vitro model.

Arteriosclerosis An Official Journal of the American Heart Association Inc ◽

10.1161/01.atv.9.4.550 ◽

1989 ◽

Vol 9 (4) ◽

pp. 550-559 ◽

Cited By ~ 40

Author(s):

E G Langeler ◽

I Snelting-Havinga ◽

V W van Hinsbergh

Keyword(s):

Endothelial Cells ◽

In Vitro Model ◽

Low Density Lipoproteins ◽

Low Density ◽

Vasoactive Substances ◽

Vitro Model ◽

Arterial Endothelial Cells

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Platelets and Endothelial Cells Act in Concert to Delay Thrombolysis – Evidence from an In Vitro Model of the Human Occlusive Thrombus

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614953 ◽

1998 ◽

Vol 79 (03) ◽

pp. 602-608 ◽

Cited By ~ 3

Author(s):

W. G. Jerome ◽

S. Handt ◽

R. R. Hantgan

Keyword(s):

Endothelial Cells ◽

Plasminogen Activator ◽

In Vitro Model ◽

Umbilical Vein ◽

Plasminogen Activator Inhibitor ◽

Genetically Engineered ◽

Cellular Mechanisms ◽

Vitro Model ◽

Activator Inhibitor

SummaryThe molecular and cellular mechanisms that over a period of hours render a human thrombus progressively resistant to fibrinolysis have been probed with a novel in vitro model. The kinetics of clot formation and fibrinolysis were monitored by laser light scattering with platelet-rich model thrombi contained in cylindrical flow chambers. In selected experiments, human umbilical vein endothelial cells were also cultured to confluence on the inner walls of these “glass blood vessels”. Following an “aging” period (0.5, 2 or 4 h), each thrombus was gently perfused with a bolus of plasminogen/recombinant tissue plasminogen activator to induce fibrinolysis. Platelets delayed lysis of 2 h-aged thrombi by ~70% and (non-stimulated) endothelial cells by ~30%, compared to cell-free control clots. However, even greater lytic delays (~260%) resulted when both vascular cells were present in the same 2 h-aged thrombus. In contrast, rapid lysis was consistently achieved with R298E,R299E t-PA, a genetically engineered plasminogen activator that is insensitive to inhibition by plasminogen activator inhibitor type 1. These observations suggest platelets and endothelial cells act in concert to enrich the fibrin scaffold of an aging human thrombus in plasminogen activator inhibitor. We propose that the presence of both platelets and endothelial cells may contribute to progressive thrombolytic resistance.

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Screening of Anti-Hypoxia/Reoxygenation Agents by an in vitro Model. Part 1: Natural Inhibitors for Protein Tyrosine Kinase Activated by Hypoxia/Reoxygenation in Cultured Human Umbilical Vein Endothelial Cells

Planta Medica ◽

10.1055/s-2000-11128 ◽

2000 ◽

Vol 66 (2) ◽

pp. 114-118 ◽

Cited By ~ 29

Author(s):

You-Wei Zhang ◽

Ikuo Morita ◽

Gang Shao ◽

Xin-Sheng Yao ◽

Sei-itsu Murota

Keyword(s):

Endothelial Cells ◽

Tyrosine Kinase ◽

Protein Tyrosine Kinase ◽

In Vitro Model ◽

Umbilical Vein ◽

Vitro Model ◽

Umbilical Vein Endothelial Cells ◽

Natural Inhibitors ◽

Hypoxia Reoxygenation

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Monocytes Maintain Tissue Factor Activity after Cytolysis of Bacteria‐Infected Endothelial Cells in an In Vitro Model of Bacterial Endocarditis

The Journal of Infectious Diseases ◽

10.1086/344231 ◽

2002 ◽

Vol 186 (8) ◽

pp. 1145-1154 ◽

Cited By ~ 13

Author(s):

Marcel H. A. M. Veltrop ◽

Henry Beekhuizen

Keyword(s):

Endothelial Cells ◽

Tissue Factor ◽

In Vitro Model ◽

Bacterial Endocarditis ◽

Factor Activity ◽

Tissue Factor Activity ◽

Vitro Model

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Venular endothelial cells from bovine heart

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1988.254.6.h1211 ◽

1988 ◽

Vol 254 (6) ◽

pp. H1211-H1217 ◽

Cited By ~ 9

Author(s):

M. E. Schelling ◽

C. J. Meininger ◽

J. R. Hawker ◽

H. J. Granger

Keyword(s):

Endothelial Cells ◽

In Vitro Model ◽

Protein Transport ◽

Culture Conditions ◽

Support Cell ◽

Vitro Model ◽

Microvascular Cells ◽

Selection Of ◽

Coronary Angiogenesis

Coronary venular endothelial cells were isolated by a bead-perfusion technique that allowed the selection of endothelial cells from venules of a specific size. Culture conditions for the microvascular cells were established. Cells grew well in supplemented Dulbecco's modified Eagle's medium. The effect of various substrata on the proliferation of the venular endothelial cells was determined. Matrigel, gelatin, and fibronectin supported high levels of proliferation. Cell shape was correlated with ability of the substratum to support cell proliferation. Cells exhibiting a broad, flattened morphology achieved high levels of proliferation. The formation of vessel meshworks by the coronary venular endothelial cells provides an in vitro model for the study of coronary angiogenesis. Confluent monolayers of these cells can be utilized to examine mechanisms of water and protein transport across coronary venules.

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