Immunological Control of Ocular Herpetic Infections

眼部疱疹感染的免疫控制

• 批准号: 12470365
• 负责人: INOUE Yoshitsugu
• 金额: $ 7.23万
• 依托单位: Tottori University (2001-2002)Osaka University (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470365/
• 关键词: herpetic keratitis; T cells; herpes simplex virus; DNA vaccine; gD; gD-IL-2; cytokines; CD4 cells; CD4細胞

1. Efficacy of gD against murine herpetic keratitis - DNA vaccine and protein vaccineWe 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. OthersImmunohistochemical 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.

1. 单纯疱疹1型（HSV-1）糖蛋白D （gD）与人白细胞介素-1 （IL-2）组成的融合蛋白D （gD-IL-2）接种小鼠疱疹性角膜炎DNA疫苗和蛋白疫苗的效果。并从质粒DNA和蛋白两方面比较评价了gD-IL-2对小鼠疱疹性角膜炎的疗效。构建含有gD-IL-2的质粒，纯化gD-IL-2肽。给BALB/c小鼠皮下或结膜下注射两次0.001mg/0.1ml的gD-IL-2肽，或皮下或结膜下注射两次0.09mg/0.05ml的gD-IL-2质粒DNA。检测抗HSV-1的中和抗体滴度和延迟型超敏反应（DTH）。用CHR3株HSV-1经角膜刺激免疫小鼠。对上皮性和间质性角膜炎的临床表现进行评分。结果表明，gD-IL-2肽或gD-IL-2 dna免疫小鼠间质性角膜炎受到抑制，而上皮性角膜炎则无抑制作用。证实质粒gD-IL-2可引起显著的血清病毒中和效价和DTH反应。对于gD-IL-2质粒DNA，也评价了滴注的效果。DNA疫苗（含0.09mg/0.01ml滴眼液）局部免疫可引起抗HSV-1的中和抗体和DTH反应，并可引起抗hsv感染细胞的细胞毒性T细胞活性。这种疗法完全阻止了疱疹性间质角膜炎的发展。此外，还进行了小鼠疱疹性角膜炎、特应性皮炎患者疱疹性角膜炎的免疫组化研究，以及疱疹性角膜炎患者衍生病毒株的流行病学研究。

项目成果

Nakata K, Inoue Y, Harada J, Maeda N, Watanabe H, Tano Y, Shimomura Y, Harino S, Sawa M: "A high incidence of Staphylococcus aureus colonization in the external eyes of patients with atopic dermatitis"Ophthalmology. 107. 2167-2171 (2000)

Nakata K、Inoue Y、Harada J、Maeda N、Watanabe H、Tano Y、Shimomura Y、Harino S、Sawa M：“特应性皮炎患者外眼金黄色葡萄球菌定植的发生率很高”眼科。

Inoue T, Inoue Y et al.: "Immunohistological study of infiltrated cells and cytokines in murine herpetic keratitis"Acta Opthalmol Scand. 79. 484-487 (2001)

Inoue T、Inoue Y 等人：“小鼠疱疹性角膜炎浸润细胞和细胞因子的免疫组织学研究”Acta Opthalmol Scand。

Inoue T, Inoue Y et al.: "Topical administration of HSV gD-IL-2 DNA is highly protective against murine herpetic stromal keratitis"Cornea. 21. 106-110 (2002)

Inoue T、Inoue Y 等人：“局部施用 HSV gD-IL-2 DNA 对小鼠疱疹性基质角膜炎具有高度保护作用”角膜。

Vmenek, Inoue Y et al.: "Genotyping of herpes simplex virus type1 strains isolated from ocular materials of patients with herpetic keratitis"J Med Virol. 70(in press). (2003)

Vmenek、Inoue Y 等人：“从疱疹性角膜炎患者的眼部材料中分离出的 1 型单纯疱疹病毒株的基因分型”J Med Virol。

井上幸次 他: "角膜クリニック(第2版)"医学書院. 358 (2003)

井上浩司等人：“角膜诊所（第2版）”Igaku Shoin 358（2003）。