Retinoic acid‐inducible gene‐I, melanoma differentiation‐associated gene 5 and C‐X‐C motif chemokine ligand 10 are induced by a Toll‐like receptor 3 agonist in human brain microvascular endothelial cells

Imaizumi, Tadaatsu; Arai, Akine; Kawaguchi, Shogo; Hayakari, Ryo; Matsumiya, Tomoh; Seya, Kazuhiko; Yoshida, Hidemi; Tanaka, Hiroshi

doi:10.1111/cen3.12463

Cited by 8 publications

(5 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, the regulatory mechanisms of CXCL10 production in RFLS have not been fully characterized. The TLR3/IFN-β/MDA5/CXCL10 axis demonstrated by our study corroborates previous findings of the role of MDA5 in regulating CXCL10 expression in glomerular mesangial cells [ 15 ], brain microvascular endothelial cells [ 26 ], and hepatocellular carcinoma cells [ 27 ]. The present study revealed a new role of MDA5 in RA synoviocytes and offers new insights about RA sterile inflammation mechanisms.…”

Section: Discussionsupporting

confidence: 91%

Role of MDA5 in regulating CXCL10 expression induced by TLR3 signaling in human rheumatoid fibroblast-like synoviocytes

et al. 2021

Self Cite

View full text Add to dashboard Cite

C-X-C motif chemokine 10 (CXCL10) is an inflammatory chemokine and a key molecule in the pathogenesis of rheumatoid arthritis (RA). Melanoma differentiation-associated gene 5 (MDA5) is an RNA helicase that plays a role in innate immune and inflammatory reactions. The details of the regulatory mechanisms of CXCL10 production and the precise role of MDA5 in RA synovitis have not been fully elucidated. The aim of this study was to examine the role of MDA5 in regulating CXCL10 expression in cultured human rheumatoid fibroblast-like synoviocytes (RFLS). RFLS was stimulated with Toll-like receptor 3 (TLR3) ligand polyinosinic:polycytidylic acid (poly I:C), a synthetic double-stranded RNA mimetic. Expression of interferon beta (IFN-β), MDA5, and CXCL10 was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, and enzyme-linked immunosorbent assay. A neutralizing antibody of IFN-β and siRNA-mediated MDA5 knockdown were used to determine the role of these molecules in regulating CXCL10 expression downstream of TLR3 signaling in RFLS. Poly I:C induced IFN-β, MDA5, and CXCL10 expression in a concentration- and time-dependent manner. IFN-β neutralizing antibody suppressed the expression of MDA5 and CXCL10, and knockdown of MDA5 decreased a part of CXCL10 expression (p < 0.001). The TLR3/IFN-β/CXCL10 axis may play a crucial role in the inflammatory responses in RA synovium, and MDA5 may be partially involved in this axis.

show abstract

Section: Discussionsupporting

confidence: 91%

Role of MDA5 in regulating CXCL10 expression induced by TLR3 signaling in human rheumatoid fibroblast-like synoviocytes

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…ZAPS, but not ZAPL, enhances RIG-I-mediated signaling in HEK293 cells after viral infection [7]. We have previously reported that RIG-I is involved in CXCL10 expression induced by TLR3 signaling in hCMEC/D3 cells [19]. However, ZAP knockdown did not affect poly ICinduced expression of RIG-I and CXCL10 in hCMEC/D3 cells in the present study.…”

Section: Discussion/conclusioncontrasting

confidence: 74%

“…We have previously reported that cultured hCMEC/D3 cells express TLR3 [18]. Adding poly IC to culture medium induces the expression of IFN-β, a type I IFN, via activation of IRF3 and NF-κB, and newly synthesized IFN-β is involved in RIG-I and CXCL10 expression in hCMEC/D3 cells [19]. The aim of this study was to investigate ZAP expression in cultured hCMEC/D3 cells treated with poly IC.…”

Section: Introductionmentioning

confidence: 99%

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

Okudera¹,

Odawara²,

Arakawa³

et al. 2021

Neuroimmunomodulation

View full text Add to dashboard Cite

Introduction: 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). Methods: 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. Results: 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. Conclusion: 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.

show abstract

“…IFN-b is a key cytokine in innate immune reactions in the downstream of TLR3. 19 We have previously reported that the treatment of hCMEC/D3 cells with poly IC induces the expression of CCL5 in an IFN-b-independent manner, 15 whereas CXCL10 20 and ISG60 17 are induced by poly IC in an IFN-b-dependent manner. Thus, poly IC induces the expression of various genes either in an IFN-bindependent or -dependent manner.…”

Section: Discussionmentioning

confidence: 98%

Induction of C‐C motif chemokine ligand 2 through Toll‐like receptor 3 signaling in human cerebral microvascular endothelial cell/D3 cells: possible regulation by nuclear factor‐κB

Sassa

Hirono

Shiratori

et al. 2019

Clinical & Exp Neuroim

Self Cite

View full text Add to dashboard Cite

Objective C-C motif chemokine ligand 2 (CCL2) is a potent chemokine that plays an important role in host defense and inflammatory diseases in the central nervous system. Toll-like receptor 3 (TLR3) is a pattern recognition receptor that recognizes viral double-stranded RNAs. In the present study, we examined whether a TLR3 agonist induces CCL2 expression in brain microvascular endothelial cells, and investigated the mechanism underlying upregulation of CCL2 expression through TLR3 signaling. Methods A human brain microvascular endothelial cell line, human cerebral microvascular endothelial cell (hCMEC)/D3 was treated with a TLR3 agonist, polyinosinic-polycytidylic acid (poly IC). The mRNA and protein expression of CCL2 were evaluated using real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The role of TLR3 and nuclear factor-jB (NF-jB) were examined with RNA interference. We also examined the effect of pretreatment with an NF-jB inhibitor, SN50, an interferon regulatory factor 3 inhibitor MR67307 and human type I interferon neutralizing antibody mixture on poly IC-induced CCL2 expression. Results Expression of CCL2 was induced by poly IC. Poly IC-induced CCL2 expression was decreased by knockdown of TLR3 or NF-jB p65 and by pretreatment with SN50. Neither MR67307 nor human type I interferon neutralizing antibody mixture affected the CCL2 expression induced by poly IC. Conclusions CCL2 expression was induced by poly IC through TLR3 signaling, and NF-jB is involved in this reaction. CCL2 produced by human brain endothelial cells might induce the infiltration of monocytes followed by immune and inflammatory reactions in the brain, and could be involved in the pathogenesis of viral encephalitis.

show abstract

Retinoic acid‐inducible gene‐I, melanoma differentiation‐associated gene 5 and C‐X‐C motif chemokine ligand 10 are induced by a Toll‐like receptor 3 agonist in human brain microvascular endothelial cells

Cited by 8 publications

References 24 publications

Role of MDA5 in regulating CXCL10 expression induced by TLR3 signaling in human rheumatoid fibroblast-like synoviocytes

Role of MDA5 in regulating CXCL10 expression induced by TLR3 signaling in human rheumatoid fibroblast-like synoviocytes

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

Induction of C‐C motif chemokine ligand 2 through Toll‐like receptor 3 signaling in human cerebral microvascular endothelial cell/D3 cells: possible regulation by nuclear factor‐κB

Contact Info

Product

Resources

About