Development and clinical application of nasal mucosa vaccine in influenza virus

流感病毒鼻粘膜疫苗的研制及临床应用

• 批准号: 14570758
• 负责人: MORI Masaaki
• 金额: $ 2.18万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570758/
• 关键词: nasal mucosa vaccine; CpG motif; influenza virus; cytokines; インフルエンザ; CpGモチーフ; インフルエンザウイルス; 経鼻投与; ワクチン

1) Mixture of virus antigen nucleus protein, rat peculiar CpG motif and the special cellulose・Extraction of the virus antigen nucleus protein :We used it in the state that inactivated influenza virus PR8 (H3N2) by formalin. Then, we extracted existing nucleus protein by liquid chromatography in this virus film and confirmed it by Western blot method and used this as virus antigen nucleus protein for vaccine making. After subcutaneous or intramuscular injection of inactivation vaccine itself or the antigen nucleus protein in F344 rats (seven-weeks old of age) as a laboratory animal, we collected blood after before and after injection regularly and measured influenza antibody titer of the sera by ELISA method and compared them. We confirmed that I examined CTL activity ability at the same time, and the instruction of the immune system of the host side was caused.・Design of rat peculiar CpG motif :It was recognized that CpG motif including the arrangement of TGACGTT raised an immunity eff … More ect most such as B cell activity, a cytokine production instruction, so we used it for an experiment. We collected sera and lymphocytes before and after injection regularly, and measured the antibody titer measurement by ELISA method, the quantity of cytokine production by ELISPOT method added to CTL activity.2) Development of the nasal mucosa vaccine in influenza virus・Examination of the reactivity of the nasal mucous membrane vaccine :We mixed virus antigen nucleus protein and CpG motif as 1:1, and added two kinds of cellulose ( hydroxypropyl cellulose (HPC) 8: microcrystal glass cellulose (MCC) 2) to it. Finally, we kept the ratio and atomized it in a nasal cavity.・Comparison with the liquid vaccine :We compared the effect between the inactivation liquid vaccine and the powdered vaccine which adjusted to the most suitable density. Actually, liquid vaccine superior to our vaccine in antibody production (IgG/IgA), HA antibody titer, both points of the cell cytotoxicity function. In this regard, it was recognized that we needed a further device to the clinical trial study of the practical use of this agent. However, it was suggested that our vaccine had the effect of 75% of intranasal and 60% of subcutaneous injection with liquid vaccine.Finally, we decided to go while adding improvement and compiled the result to the existing stage as an article (in press). And then, we hope that we contribute the study to foreign countries magazine in future. Less

1)病毒抗原核蛋白、大鼠特有的CpG基序和特殊纤维素的混合物·病毒抗原核蛋白的提取：在福尔马林灭活流感病毒PR 8（H3 N2）的状态下使用。然后，我们用液相色谱法提取了该病毒膜中存在的核蛋白，并用Western blot方法进行了鉴定，并将其作为疫苗制备中的病毒抗原核蛋白。以F344大鼠（7周龄）为实验动物，皮下或肌肉注射灭活疫苗本身或抗原核蛋白后，定期采集注射前后的血液，用ELISA法测定血清流感抗体效价，并进行比较。我们确认我同时检查了CTL活性能力，并引起了宿主方免疫系统的指令。大鼠特异性CpG基序的设计：含TGACGTT排列的CpG基序被认为具有免疫效应 ...更多信息 ect大多数如B细胞活性，一个细胞因子的产生指令，所以我们用它做了一个实验。流感病毒鼻粘膜疫苗的研制：将流感病毒抗原核蛋白与CpG基序按1：1混合，并加入两种纤维素酶，制备了流感病毒鼻粘膜疫苗，并对疫苗的免疫原性进行了检测（羟丙基纤维素（HPC）8：微晶玻璃纤维素（MCC）2）。最后，我们保持比例并将其雾化在鼻腔中。与液体疫苗的比较：将灭活液体疫苗与调整至最适浓度的粉末疫苗进行效果比较。实际上，液体疫苗在抗体产生（IgG/伊加）、HA抗体滴度、细胞毒作用这两点上都上级我们的疫苗。在这方面，人们认识到，我们需要进一步的设备，以临床试验研究的实际使用这种代理。但是，有人提出，我们的疫苗有75%的鼻内和60%的皮下注射液体疫苗的效果。最后，我们决定一边去，一边增加改进，并将结果汇编到现有阶段作为一篇文章（出版中）。同时，我们希望将来能将我们的研究贡献给国外的杂志。少

项目成果

Development of nasal mucosa vaccine in influenza virus

流感病毒鼻粘膜疫苗的研制

• 期刊: Vaccine (in press)
• 作者: Mori M;et al.
• 通讯作者: et al.

海外ワクチン雑誌に投稿準備中

准备向海外疫苗杂志投稿

• 作者: 森 雅亮