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Title: Poly(I:C) Induces Human Lung Endothelial Barrier Dysfunction by Disrupting Tight Junction Expression of Claudin-5

Abstract

Viral infections are often accompanied by pulmonary microvascular leakage and vascular endothelial dysfunction via mechanisms that are not completely defined. Here, we investigated the effect of the Toll-like receptor 3 (TLR3) ligand polyinosinic-polycytidylic acid [Poly(I:C)], a synthetic analog of viral double-stranded RNA (dsRNA) commonly used to simulate viral infections, on the barrier function and tight junction integrity of primary human lung microvascular endothelial cells. Poly(I:C) stimulated IL-6, IL-8, TNFα, and IFNβ production in conjunction with the activation of NF-κB and IRF3 confirming the Poly(I:C)-responsiveness of these cells. Poly(I:C) increased endothelialmonolayer permeability with a corresponding dose- and time-dependent decrease in the expression of claudin-5, a transmembrane tight junction protein and reduction of CLDN5 mRNA levels. Immunofluorescence experiments revealed disappearance of membrane-associated claudin-5 and co-localization of cytoplasmic claudin-5 with lysosomal-associated membrane protein 1. Chloroquine and Bay11-7082, inhibitors of TLR3 and NF-κB signaling, respectively, protected against the loss of claudin-5. Altogether, these findings provide new insight on how dsRNA-activated signaling pathways may disrupt vascular endothelial function and contribute to vascular leakage pathologies.

Authors:
 [1];  [1];  [2];  [2];  [1];  [3]
  1. Food and Drug Administration, Silver Springs, MD (United States)
  2. Food and Drug Administration, Silver Spring, MD (United States)
  3. Hungarian Academy of Sciences (Hungary)
Publication Date:
Research Org.:
Food and Drug Administration, Silver Spring, Maryland (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1377835
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 11; Journal Issue: 8; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; immune receptor signaling; cell membranes; nuclear straining; tight junctions; toll-like receptors; endothelial cells; fluorescence imaging; permeability

Citation Formats

Huang, Li -Yun, Stuart, Christine, Takeda, Kazuyo, D’Agnillo, Felice, Golding, Basil, and Deli, Mária A. Poly(I:C) Induces Human Lung Endothelial Barrier Dysfunction by Disrupting Tight Junction Expression of Claudin-5. United States: N. p., 2016. Web. doi:10.1371/journal.pone.0160875.
Huang, Li -Yun, Stuart, Christine, Takeda, Kazuyo, D’Agnillo, Felice, Golding, Basil, & Deli, Mária A. Poly(I:C) Induces Human Lung Endothelial Barrier Dysfunction by Disrupting Tight Junction Expression of Claudin-5. United States. doi:10.1371/journal.pone.0160875.
Huang, Li -Yun, Stuart, Christine, Takeda, Kazuyo, D’Agnillo, Felice, Golding, Basil, and Deli, Mária A. 2016. "Poly(I:C) Induces Human Lung Endothelial Barrier Dysfunction by Disrupting Tight Junction Expression of Claudin-5". United States. doi:10.1371/journal.pone.0160875. https://www.osti.gov/servlets/purl/1377835.
@article{osti_1377835,
title = {Poly(I:C) Induces Human Lung Endothelial Barrier Dysfunction by Disrupting Tight Junction Expression of Claudin-5},
author = {Huang, Li -Yun and Stuart, Christine and Takeda, Kazuyo and D’Agnillo, Felice and Golding, Basil and Deli, Mária A.},
abstractNote = {Viral infections are often accompanied by pulmonary microvascular leakage and vascular endothelial dysfunction via mechanisms that are not completely defined. Here, we investigated the effect of the Toll-like receptor 3 (TLR3) ligand polyinosinic-polycytidylic acid [Poly(I:C)], a synthetic analog of viral double-stranded RNA (dsRNA) commonly used to simulate viral infections, on the barrier function and tight junction integrity of primary human lung microvascular endothelial cells. Poly(I:C) stimulated IL-6, IL-8, TNFα, and IFNβ production in conjunction with the activation of NF-κB and IRF3 confirming the Poly(I:C)-responsiveness of these cells. Poly(I:C) increased endothelialmonolayer permeability with a corresponding dose- and time-dependent decrease in the expression of claudin-5, a transmembrane tight junction protein and reduction of CLDN5 mRNA levels. Immunofluorescence experiments revealed disappearance of membrane-associated claudin-5 and co-localization of cytoplasmic claudin-5 with lysosomal-associated membrane protein 1. Chloroquine and Bay11-7082, inhibitors of TLR3 and NF-κB signaling, respectively, protected against the loss of claudin-5. Altogether, these findings provide new insight on how dsRNA-activated signaling pathways may disrupt vascular endothelial function and contribute to vascular leakage pathologies.},
doi = {10.1371/journal.pone.0160875},
journal = {PLoS ONE},
number = 8,
volume = 11,
place = {United States},
year = 2016,
month = 8
}

Journal Article:
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