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 endothelial monolayer 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. Together, these findings provide new insight on how dsRNA-activated signaling pathways may disrupt vascular endothelial function and contribute to vascular leakage pathologies.
authors: Laboratory of Plasma Derivatives, Division of Hematology Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, United States of America.
authors: Li-Yun
authors: LY
authors: Huang
authors: Laboratory of Plasma Derivatives, Division of Hematology Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, United States of America.
authors: Christine
authors: C
authors: Stuart
authors: Microscopy and Imaging Core Facility, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, United States of America.
authors: Kazuyo
authors: K
authors: Takeda
authors: Laboratory of Biochemistry and Vascular Biology, Division of Hematology Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, United States of America.
authors: Felice
authors: F
authors: D'Agnillo
authors: Laboratory of Plasma Derivatives, Division of Hematology Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, United States of America.
authors: Basil
authors: B
authors: Golding
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism; D062507:Claudin-5 / Q000235:genetics*; D015536:Down-Regulation / Q000187:drug effects*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D050838:Interferon Regulatory Factor-3 / Q000378:metabolism; D008168:Lung / Q000166:cytology*; D016328:NF-kappa B / Q000378:metabolism; D010539:Permeability / Q000187:drug effects; D011070:Poly I-C / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D019108:Tight Junctions / Q000187:drug effects* / Q000378:metabolism*; D051196:Toll-Like Receptor 3 / Q000378:metabolism
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D018925:Chemokines; D062507:Claudin-5; C494232:IRF3 protein, human; D050838:Interferon Regulatory Factor-3; D016328:NF-kappa B; D012333:RNA, Messenger; C470409:TICAM1 protein, human; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D011070:Poly I-C