Expression of Zinc-Finger Antiviral Protein in hCMEC/D3 Human Cerebral Microvascular Endothelial Cells: Effect of a Toll-Like Receptor 3 Agonist

2021 ◽  
pp. 1-10
Author(s):  
Mako Okudera ◽  
Minami Odawara ◽  
Masashi Arakawa ◽  
Shogo Kawaguchi ◽  
Kazuhiko Seya ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Protein Expression ◽  
Zinc Finger ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Toll Like Receptor ◽  
Antiviral Protein ◽  
Polycytidylic Acid ◽  
Microvascular Endothelial ◽  
The Brain

<b><i>Introduction:</i></b> Invasion of viruses into the brain causes viral encephalitis, which can be fatal and causes permanent brain damage. The blood-brain barrier (BBB) protects the brain by excluding harmful substances and microbes. Brain microvascular endothelial cells are important components of the BBB; however, the mechanisms of antiviral reactions in these cells have not been fully elucidated. Zinc-finger antiviral protein (ZAP) is a molecule that restricts the infection of various viruses, and there are 2 major isoforms: ZAPL and ZAPS. Toll-like receptor 3 (TLR3), a pattern-recognition receptor against viral double-stranded RNA, is implicated in antiviral innate immune reactions. The aim of this study was to investigate the expression of ZAP in cultured hCMEC/D3 human brain microvascular endothelial cells treated with an authentic TLR3 agonist polyinosinic-polycytidylic acid (poly IC). <b><i>Methods:</i></b> hCMEC/D3 cells were cultured and treated with poly IC. Expression of ZAPL and ZAPS mRNA was investigated using quantitative reverse transcription-polymerase chain reaction, and protein expression of these molecules was examined using western blotting. The role of nuclear factor-κB (NF-κB) was examined using the NF-κB inhibitor, SN50. The roles of interferon (IFN)-β, IFN regulatory factor 3 (IRF3), tripartite motif protein 25 (TRIM25), and retinoic acid-inducible gene-I (RIG-I) in poly IC-induced ZAPS expression were examined using RNA interference. Propagation of Japanese encephalitis virus (JEV) was examined using a focus-forming assay. <b><i>Results:</i></b> ZAPS mRNA and protein expression was upregulated by poly IC, whereas the change of ZAPL mRNA and protein levels was minimal. Knockdown of IRF3 or TRIM25 decreased the poly IC-induced upregulation of ZAPS, whereas knockdown of IFN-β or RIG-I did not affect ZAPS upregulation. SN50 did not affect ZAPS expression. Knockdown of ZAP enhanced JEV propagation. <b><i>Conclusion:</i></b> ZAPL and ZAPS were expressed in hCMEC/D3 cells, and ZAPS expression was upregulated by poly IC. IRF3 and TRIM25 are involved in poly IC-induced upregulation of ZAPS. ZAP may contribute to antiviral reactions in brain microvascular endothelial cells and protect the brain from invading viruses such as JEV.

Anthropogenic Iron Oxide Nanoparticles Induce Damage to Brain Microvascular Endothelial Cells Forming the Blood-Brain Barrier

2021 ◽  
Author(s):  
Lorena Gárate-Vélez ◽  
Claudia Escudero-Lourdes ◽  
Daniela Salado-Leza ◽  
Armando González-Sánchez ◽  
Ildemar Alvarado-Morales ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Blood Brain ◽  
Fe3o4 Nps ◽  
Microvascular Endothelial ◽  
The Brain

Background: Iron nanoparticles, mainly in magnetite phase (Fe3O4 NPs), are released to the environment in areas with high traffic density and braking frequency. Fe3O4 NPs were found in postmortem human brains and are assumed to get directly into the brain through the olfactory nerve. However, these pollution-derived NPs may also translocate from the lungs to the bloodstream and then, through the blood-brain barrier (BBB), into the brain inducing oxidative and inflammatory responses that contribute to neurodegeneration. Objective: To describe the interaction and toxicity of pollution-derived Fe3O4 NPs on primary rat brain microvascular endothelial cells (rBMECs), main constituents of in vitro BBB models. Methods: Synthetic bare Fe3O4 NPs that mimic the environmental ones (miFe3O4) were synthesized by co-precipitation and characterized using complementary techniques. The rBMECs were cultured in Transwell® plates. The NPs-cell interaction was evaluated through transmission electron microscopy and standard colorimetric in vitro assays. Results: The miFe3O4 NPs, with a mean diameter of 8.45 ± 0.14 nm, presented both magnetite and maghemite phases, and showed super-paramagnetic properties. Results suggest that miFe3O4 NPs are internalized by rBMECs through endocytosis and that they are able to cross the cells monolayer. The lowest miFe3O4 NPs concentration tested induced mid cytotoxicity in terms of 1) membrane integrity (LDH release) and 2) metabolic activity (MTS transformation). Conclusion: Pollution-derived Fe3O4 NPs may interact and cross the microvascular endothelial cells forming the BBB and cause biological damage.

scholarly journals Caveolin-1 regulates expression of junction-associated proteins in brain microvascular endothelial cells

Blood ◽  
2006 ◽  
Vol 109 (4) ◽  
pp. 1515-1523 ◽  
Author(s):  
Li Song ◽  
Shujun Ge ◽  
Joel S. Pachter
Keyword(s):  
Endothelial Cells ◽  
Protein Expression ◽  
Adherens Junction ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Caveolin 1 ◽  
Synthetic Cell ◽  
Associated Proteins ◽  
Junctional Protein ◽  
Microvascular Endothelial

Abstract Recent evidence from this laboratory indicated that reduced expression of caveolin-1 accompanied the diminished expression of tight junction (TJ)–associated proteins occludin and zonula occludens-1 (ZO-1) following stimulation of brain microvascular endothelial cells (BMECs) with the chemokine CCL2 (formerly called MCP-1). Because attenuated caveolin-1 levels have also been correlated with heightened permeability of other endothelia, the objective of this study was to test the hypothesis that reduced caveolin-1 expression is causally linked to the action of CCL2 on BMEC junctional protein expression and barrier integrity. This was achieved using adenovirus to nondestructively deliver caveolin-1 siRNA (Ad-siCav-1) to BMEC monolayers, which model the blood-brain barrier (BBB). Treatment with siRNA reduced the caveolin-1 protein level as well as occludin and ZO-1. Additionally, occludin exhibited dissociation from the cytoskeletal framework. These changes were attended by comparable alterations in adherens junction (AJ)–associated proteins, VE-cadherin and β-catenin, increased BMEC paracellular permeability, and facilitated the ability of CCL2 to stimulate monocytic transendothelial migration. Furthermore, treating BMECs with cavtratin, a synthetic cell-permeable peptide encoding the caveolin-1 scaffolding domain, antagonized effects of both Ad-siCav-1 and CCL2. These results collectively highlight caveolin-1 loss as a critical step in CCL2-induced modulation of BMEC junctional protein expression and integrity, and possibly serve a crucial role in regulating inflammation at the BBB.

scholarly journals Brain Microvascular Endothelial Cells-Derived HMGB1 Facilitates Monocyte Transendothelial Migration Favoring JEV Neuroinvasion

2020 ◽  
Author(s):  
Song-Song Zou ◽  
Qing-Cui Zou ◽  
Wen-Jing Xiong ◽  
Ning-Yi Cui ◽  
Ke Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Japanese Encephalitis ◽  
Immune Cells ◽  
Transendothelial Migration ◽  
Trojan Horse ◽  
Microvascular Endothelial Cells ◽  
High Morbidity ◽  
Brain Microvascular Endothelial Cells ◽  
Microvascular Endothelial ◽  
The Brain

Abstract Background: Infection with the Japanese encephalitis virus (JEV) induced high morbidity and mortality, even caused permanent neurological sequelae. However, the pathways and mechanisms of JEV invasion into the central nervous system (CNS) remain elusive. It is confirmed that extracellular HMGB1 facilitates immune cells transendothelial migration. Furthermore, it is observed that the migration of immune cells into the CNS dramatically increased during JEV infection which may benefit to viral clearance, but paradoxically accompanied by the expedite onset of Japanese encephalitis (JE) in advance. Thus, exploration of JEV neuroinvasion pathways is important for pathogenesis and prevention of JE.Methods: Brain microvascular endothelial cells were utilized for the detection of HMGB1 release in vitro. The blood-brain barrier (BBB) monolayer model (brain microvascular endothelial cells) and recombinant HMGB1 were applied for the measurement of endothelial cell activation and cells adhesion, the integrity of the BBB and the interaction with the immune cells. A genetically modified JEV expressing EGFP (EGFP-JEV) was used to trace the transmigration of JEV-infected immune cells crossing the BBB to mimic the process of neuroinfection.Results: JEV has the characteristic of neurotropism, causing HMGB1 released from BMEC and increasing adhesion molecules. BEMC-derived HMGB1 enhances leukocyte-endothelium adhesion, facilitating the transendothelial migration of JEV-infected monocytes across the BBB entry into the CNS. Thus, JEV successfully utilized the monocyte as a “Trojan horse” to spread the virus to the brain, expanding the brain infection, leading the acceleration of JE onset.Conclusion: JEV-infected monocytes, acting as “Trojan horse”, migrate to the brain, which was facilitated by BMEC-derived HMGB1, contributing to JEV neuroinvasion, and leading neuroinflammation and pathological changes of JE.

scholarly journals Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro

PLoS ONE ◽  
2012 ◽  
Vol 7 (9) ◽  
pp. e45977 ◽  
Author(s):  
Johanna P. Laakkonen ◽  
Tatjana Engler ◽  
Ignacio A. Romero ◽  
Babette Weksler ◽  
Pierre-Olivier Couraud ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Adenovirus Vectors ◽  
Microvascular Endothelial ◽  
The Brain

scholarly journals Cerebrovascular plasticity: Processes that lead to changes in the architecture of brain microvessels

2019 ◽  
Vol 39 (8) ◽  
pp. 1413-1432 ◽  
Author(s):  
Max I Bogorad ◽  
Jackson G DeStefano ◽  
Raleigh M Linville ◽  
Andrew D Wong ◽  
Peter C Searson
Keyword(s):  
Endothelial Cells ◽  
Neural Activity ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Hierarchical Network ◽  
Brain Microvessels ◽  
Endothelial Cell Death ◽  
Microvascular Endothelial ◽  
Development And Aging ◽  
The Brain

The metabolic demands of the brain are met by oxygen and glucose, supplied by a complex hierarchical network of microvessels (arterioles, capillaries, and venules). Transient changes in neural activity are accommodated by local dilation of arterioles or capillaries to increase cerebral blood flow and hence nutrient availability. Transport and communication between the circulation and the brain is regulated by the brain microvascular endothelial cells that form the blood–brain barrier. Under homeostatic conditions, there is very little turnover in brain microvascular endothelial cells, and the cerebrovascular architecture is largely static. However, changes in the brain microenvironment, due to environmental factors, disease, or trauma, can result in additive or subtractive changes in cerebrovascular architecture. Additions occur by angiogenesis or vasculogenesis, whereas subtractions occur by vascular pruning, injury, or endothelial cell death. Here we review the various processes that lead to changes in the cerebrovascular architecture, including sustained changes in the brain microenvironment, development and aging, and injury, disease, and repair.

Cerebral microvascular endothelial cell Na-K-Cl cotransport: regulation by astrocyte-conditioned medium

1995 ◽  
Vol 268 (3) ◽  
pp. C747-C754 ◽  
Author(s):  
M. E. O'Donnell ◽  
A. Martinez ◽  
D. Sun
Keyword(s):  
Endothelial Cells ◽  
Conditioned Medium ◽  
Transport Systems ◽  
Microvascular Endothelial Cells ◽  
Asymmetric Distribution ◽  
Brain Microvascular Endothelial Cells ◽  
Cyclic Monophosphate ◽  
Microvascular Endothelial ◽  
The Brain ◽  
Astrocyte Conditioned Medium

Brain microvascular endothelial cells play an important role in regulation of ion and fluid movement between the blood and the brain interstitium. Astrocytes have been shown to induce blood-brain barrier properties in the endothelial cells, including formation of tight junctions and increased expression and asymmetric distribution of enzymes and ion transport systems. Previous studies have demonstrated that endothelial cells of bovine aorta possess a highly active Na-K-Cl cotransport system that participates in intracellular volume regulation. The present study was conducted to evaluate Na-K-Cl cotransport activity of cerebral microvascular endothelial cells and to determine whether astrocyte-conditioned medium (CM) influences Na-K-Cl cotransport activity of these cells. We found the brain microvascular endothelial cells to exhibit a robust Na-K-Cl cotransport activity, comprising 50% of the total K influx. Activity of the cotransporter was stimulated by agents that elevate intracellular Ca and by hypertonicity and was inhibited by agents that elevate adenosine 3',5'-cyclic monophosphate, guanosine 3',5'-cyclic monophosphate, or activate protein kinase C. Exposure of the cells to primary astrocyte- or C6 glial cell-CM but not A7r5 or A10 vascular smooth muscle cell-CM also increased cotransport activity. However, this effect required > 1 h of exposure to CM, was additive with the effects of vasopressin, calcium ionophore, and hypertonicity, and was blocked by the protein synthesis inhibitor cycloheximide.(ABSTRACT TRUNCATED AT 250 WORDS)

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