| Project/Area Number |
12470365
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | Tottori University (2001-2002)
Osaka University (2000) |
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Principal Investigator |
INOUE Yoshitsugu Tottori University, Faculty of Medicine, Professor, 医学部, 教授 (10213183)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Hitoshi Osaka University, Medical School, Associate Professor, 大学院・医学系研究科, 助教授 (60252673)
NAKASO Nami Tottori University, Faculty of Medicine, Assistant Professor, 医学部附属病院, 助手 (90335539)
MIYAZAKI Dai Tottori University, Faculty of Medicine, Assistant Professor, 医学部附属病院, 助手 (30346358)
HAYASHI Kozaburo Kobe Institute of Medicine, Director, 所長(研究職) (20012726)
OHGURO Nobuyuki Osaka University, Medical School, Assistant Professor, 大学院・医学系研究科, 助手 (00303967)
馬場 高志 鳥取大学, 医学部, 助手 (40304216)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥11,300,000 (Direct Cost: ¥11,300,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2000: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | herpetic keratitis / T cells / herpes simplex virus / DNA vaccine / gD / gD-IL-2 / cytokines / CD4 cells / CD4細胞 |
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| Research Abstract |
1. Efficacy of gD against murine herpetic keratitis - DNA vaccine and protein vaccine
We evaluated the effect of vaccination with fusion protein D (gD-IL-2) consisting of herpes simplex type 1 (HSV-1) glycoprotein D (gD) and human interleukin-1 (IL-2). The efficacy against murine herpetic keratitis was also comparatively evaluated on both plasmid DNA and protein of gD-IL-2. Plasmid containinggD-IL-2 was constructed, and gD-IL-2 peptide was purified. BALB/c mice were injected twice hypodermally or subconjunctivally with 0.001mg/0.1ml of gD-IL-2 peptide, or twice hypodermally or subconjunctivally with 0.09mg/0.05ml of gD-IL-2 plasmid DNA. Neutralizing antibody titer and delayed-type hypersensitivity (DTH) against HSV-1 was measured. Immunized mice were challenged with CHR3 strain of HSV-1 via the cornea. The clinical picture of epithelial and stromal keratitis was scored. As the results, stromal keratitis was inhibited in gD-IL-2 peptide- or gD-IL-2 DNA-immunized mice, but epithelial keratitis was not. It was confirmed that plasmid gD-IL-2 elicited significant serum virus neutralizing titer and DTH response. Regarding gD-IL-2 plasmid DNA, the efficacy of instillation was also evaluated. Topical immunization with a DNA vaccine (eyedrops containing 0.09mg/0.01ml) elicited neutralizing antibody and DTH reaction against HSV-1, and cytotoxtic T cell activity against HSV-infected cells. This regime totally prevented the development of herpetic stromal keratitis.
2. Others
Immunohistochemical study of murine herpetic keratitis, herpetic keratitis in atopic dermatitis patients, and epidemiological study of viral strains derived from herpetic keratitis patients were also conducted.
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