| Project/Area Number |
21590967
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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 |
Respiratory organ internal medicine
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| Research Institution | Kyushu University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
FUKUYAMA Satoru 九州大学, 病院呼吸器科, 助教 (50380530)
INOUE Hiromasa 鹿児島大学, 大学院・医歯薬総合研究科呼吸器内科学, 教授 (30264039)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2009: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 閉塞性肺疾患 / COPD / 気管支喘息 / 共刺激分子 / CD86 / siRNA / マウス / 樹状細胞 / 核酸医学 / 好酸球 |
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| Research Abstract |
In COPD and asthma, the signalings among a variety of inflammatory cells and tissue-structural cells play a pivotal role in the development of diseases. These signalings are partly mediated via interaction with costimulatory molecules. The B7-family molecule CD86, expressed on the surface of dendritic cells(DCs), delivers costimulatory signals for the interaction with T cells. Using small interfering RNA(siRNA)-based approach, we investigated the therapeutic effect of CD86 knockdown on a mouse model of asthma. We prepared bone marrow-derived dendritic cells(BMDCs). CD4+ cells were isolated from the spleens of DO11. 10 mice by magnetic beads sorting method(MACS), cocultured with antigen-presenting cells, recombinant IL-4, and anti-IL-12 mAb, and then used as OVA-specific Th2 cells. BMDCs were transfected with CD86 siRNA or control siRNA, pulsed with OVA peptides and LPS, and then cocultured with Th2 cells for 24hr. The culture supernatant was collected for cytokine ELISA. The levels of IL-4, IL-5, and IL-13 in culture supernatant were significantly reduced by CD86 siRNA treatment of BMDCs. Next, we tested the therapeutic effects of CD86 siRNA for OVA-induced mouse model of asthma. CD86 siRNA or control siRNA was administrated via the intratracheal route during the OVA challenge. In microscopic study, The fluorescent-probed siRNA was successfully deposited beneath airway submucosa. The administration of CD86 siRNA significantly decreased eosinophil number and levels of IL-5 and IL-13 in the bronchoalveolar fluid, airway hyperreactivity to inhaled acetylcholine, and a level of OVA-specific IgE in the serum. These findings suggest that siRNA treatment targeting airway DCs might be of value in the development of new nucleic acid therapies for obstructive lung diseases.
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