| Project/Area Number |
17591102
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Sapporo Medical University |
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Principal Investigator |
NAGAI Kazushige Sapporo Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (50347168)
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| Co-Investigator(Kenkyū-buntansha) |
TSUTSUMI Hiroyuki Sapporo Medical University, School of Medicine, Professor, 医学部, 教授 (80217348)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Virus / Infectious Diseases / Immunology |
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| Research Abstract |
Objectives The objective of this study was to investigate antiviral activity of Toll-like receptor 3 (TLR3) against human respiratory syncytial virus (RSV). In the first year, human TLR3 gene was transiently expressed in monkey kidney COS-7 cells and anti-RSV effect was compared with control cells. In the second year, the anti-RSV effect was examined in HeLa cells with or without addition of double-stranded RNA, a ligand of TLR3.
Materials and Methods COS-7 cells were transfected with either pUNO-hTLR3-HA (Invivogen) or pUNO-HA and infected with the Long stain of RSV 24 h after the transfection. Viral RNA of the Long strain was quantified by real-time RT-PCR (Applied Biosystems) in the cell culture supernatants 24 h after the infection. HeLa cells were incubated with poly(I : C) (Amersham Biosciences), a synthetic double-stranded RNA, for 24 h then infected with the Long strain. Viral RNA of the Long strain as well as mRNA expression of interferon (IFN)-β in the cell culture supernatants and the infected cells, respectively were quantified by real-time RT-PCR.
Results The quantity of RSV in the supernatants of COS-7 cells expressing human TLR3 was significantly decreased comparing to that of the control cells. Likewise, the quantity of RSV in the supernatants of RSV-infected HeLa cells with poly(I : C) was significantly de creased comparing to that of RSV-infected HeLa cells without poly(I : C). Interestingly, mRNA expression of IFN-β was also decreased in poly(I : C)-treated, RSV-infected cells comparing to poly(I : C)-non-treated, RSV-infected cells. Discussion We have demonstrated the anti-RSV effect of TLR3 in this study. This anti-RSV effect of TLR3 may be exerted independent of IFN-β induction. Further investigation is necessary to elucidate molecular mechanisms of antiviral effects of TLR3.
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