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Expression of CCL 5 is induced by polyinosinic : polycytidylic acid in cultured hCMEC /D3 human brain microvascular endothelial cells
Author(s) -
Arai Akine,
Yoshida Hidemi,
Hayakari Ryo,
Matsumiya Tomoh,
Kawaguchi Shogo,
Seya Kazuhiko,
Tanaka Hiroshi,
Imaizumi Tadaatsu
Publication year - 2017
Publication title -
clinical and experimental neuroimmunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.297
H-Index - 15
ISSN - 1759-1961
DOI - 10.1111/cen3.12416
Subject(s) - chemokine , microbiology and biotechnology , gene knockdown , endothelial stem cell , signal transduction , transcription factor , blood–brain barrier , innate immune system , chemistry , biology , immune system , immunology , apoptosis , biochemistry , central nervous system , endocrinology , in vitro , gene
Objective Brain microvascular endothelial cells are one of the cell types that form the blood–brain barrier, and play an important role in the defense system of the brain. Toll‐like receptor 3 ( TLR 3) is a pattern‐recognition receptor against double‐stranded RNA , and TLR 3 signaling is important in antiviral innate immune reactions. However, TLR 3 signaling in brain microvascular endothelial cells is not well understood. We aimed to investigate the role of TLR 3 signaling in chemokine CCL 5 production in human brain microvascular endothelial cells. Methods The hCMEC /D3 human brain microvascular endothelial cells were cultured, and treated with an authentic TLR 3 agonist polyinosinic : polycytidylic acid. The expression of CCL 5 was examined using quantitative real‐time reverse transcription polymerase chain reaction and enzyme‐linked immunosorbent assay. Involvement of TLR 3, interferon ( IFN )‐β, or IFN ‐λ1, IFN ‐regulatory factor 3 or nuclear factor‐κB p65 in this reaction was examined using RNA interference. Translocation of p65 into the nucleus was examined using immunofluorescence staining. Results Treatment of cells with polyinosinic : polycytidylic acid induced the expression of CCL 5, IFN ‐β and IFN ‐λ1, and also the translocation of p65 into the nucleus. Knockdown of TLR 3, IFN ‐regulatory factor 3 or p65 inhibited the induction of these molecules, while knockdown of neither IFN ‐β nor IFN ‐λ1 affected the expression of CCL 5. Conclusions TLR 3 activation by polyinosinic : polycytidylic acid induces the expression of CCL 5 in cultured hCMEC /D3 cells, and IFN ‐regulatory factor 3 and p65 are involved in this reaction. CCL 5 induced by TLR 3 signaling in brain microvascular endothelial cells might contribute to antiviral protective reactions and/or detrimental responses associated with viral infection in the brain.