Primary Study of the Universal Influenza Vaccine that Images Viral Receptor

病毒受体成像通用型流感疫苗的初步研究

• 批准号: 01880021
• 负责人: SUZUKI Yasuo
• 金额: $ 2.56万
• 依托单位: University of Shizuoka, School of Pharmaceutical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B).
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01880021/
• 关键词: Influenza Virus; Receptor; Hemagglutinin; Neuraminidase; Ganglioside; Monoclonal Antibody; Vaccine; Antiidiotype antiody; インフルエンザワクチン; ノイラミニダ-ゼ; モノクロ-ナル抗体; 抗イディオタイプ抗体

This project is aimed to study primarily the developing the universal influenza vaccine, which is widely effective, independent of the variation of the viral hemagglutinin gene in relation to the anti-idiotypic antibody against the antibody directed to the influenza virus receptor.During the course of this study, following new results were obtained.1) A new simple and improved assay method to determine the receptor specificity of influenza virus variant has been developed. This method is based on the binding of the virus to the sialoglycosphingolipids (gangliosides) or sialogly-coprotein developed in silica gel thin layer plate or film blotted from the gel after electrophoresis.2) The simple method for the fractionation and isolation of receptor gangliosides for influenza viruses was developed. This method is based on the mapping the ganglioside species by chromatography using a new anion exchange beads (Q-sepharose).3) ALL the base sequences of the gene and the receptor specificity of … More viral hemagglutinin subtype (H1 - H13) were determined. It was found that NeuAcalpha2, 3 (6) Galbeta1, 3 (6) GlcNAcbeta1- is the most active receptor determinant which is commonly recognized by all the hemagglutin subtypes. The recogn ition of the hemadgglutinin the linkage between NeuAc and Gal (alpha2, 3 and alpha2, 6) was not always depend on the amino acid 226 (Leu or Gln) in the receptor binding pocket in the hemagglutinin molecule.4) Single amino acid substitution of Ser (205) located distant from the receptor binding pocket, antigenic site D of A/Mem/1/71-Bel/42 (H3N1), to Tyr changed the receptor binding specificity from 2, 3 to 2, 6. Ser (205) is located across the subunit interface from the receptor-binding site. The bultky hydrophobic tyr in the variant may cause a conformational change in the receptor-binding pocket on the neighboring subunit and influence the receptor binding.5) A new monoclonal antibody (NS24) directed to sialylparagloboside (NeuAcalpha2, 3Galbeta1, 4GlcNAcbeta1, 3Galbeta1, 4Glcbeta1-Ceramide), a common receptor gangioside of influenza A viruses was prepared. This antibody was highly specific toNeuAc2, 3Gal1, 4GlcNAc1- residue of sialylparagloboside and inhibit the binding of the A virus to erythrocytes which had been treated with sialidase and then reconstituted with sialylparagloboside. This antibody may be useful as an universal influenza vaccine effective to the wide variety of subtypes of influenza virus hemagglutinin.6) The strategy for the developing the universal influenza virus vaccine was discussed in relation to the anti-idiotypic antibody against the antibody directed to the influenza virus receptor, sialylparagloboside.7) A new common competitive inhibitor of influenza virus neuraminidase, ganglioside which contained thio-glycoside linkage between NeuAc and Gal was found. Less

本项目旨在初步研究通用流感疫苗的研制，该疫苗不受病毒血凝素基因变异与针对流感病毒受体抗体的抗独特型抗体的影响。在研究过程中，获得了以下新结果：1)建立了一种新的简单、改进的检测流感病毒变异株受体特异性的方法。这种方法是基于病毒与硅胶薄层平板或薄膜中形成的唾液酸鞘糖脂(神经节苷脂)或唾液酸辅蛋白结合，经电泳后印迹。2)建立了流感病毒受体神经节苷脂的简便分离方法。该方法基于一种新的阴离子交换小球(Q-Sepharose)的层析定位神经节苷脂物种。3)基因的所有碱基序列和…的受体特异性检测出更多的病毒血凝素亚型(H1-H13)。研究发现，NeuAcalpha2，3(6)Galbeta1，3(6)GlcNAcbeta1-是所有血凝素亚型共同识别的最活跃的受体决定簇。NeuAc和Gal(Alpha2，3和Alpha2，6)之间的血凝素的识别并不总是依赖于血凝素分子中受体结合口袋中的氨基酸226(Leu或Gln)。4)远离受体结合口袋的单一氨基酸取代A/Mem/1/71-Bel/42(H3N1)的抗原D位到Tyr，使受体结合特异性从2，3变为2，6。Ser(205)位于受体结合部位的亚基界面上。5)制备了针对甲型流感病毒常见受体神经节苷脂(NeuAcalpha2，3Galbeta1，4GlcNAcbeta1，3Galbeta1，4Glcbeta1-Ceramine)的新型单抗(NS24)。该抗体对唾液酸副糖苷的NeuAc2，3Gal1，4GlcNAc1-残基具有高度特异性，并能抑制A病毒与唾液酸酶处理后与唾液酸侧副糖苷重组的红细胞的结合。6)结合针对流感病毒受体的抗独特型抗体，探讨了开发通用流感病毒疫苗的策略。7)发现了一种新的流感病毒神经氨酸酶的竞争性抑制剂，神经节苷脂，它含有NeuAc和Gal之间的硫苷连接。较少

项目成果

Yasuo Suzuki: "Recognition of receptor sialylglycocojugates by influenza virus hemagglutinin" IXth International Carbohydrate syposium (August 12ー17,1990,Yokohama,Japan),Abstract Book pp.130(1990). 130 (1990)

Yasuo Suzuki：“流感病毒血凝素对受体唾液酸糖缀合物的识别”第 IX 届国际碳水化合物研讨会（1990 年 8 月 12-17 日，日本横滨），摘要书第 130 页（1990 年）。

鈴木康夫: "O__ーーアセチル化シアル酸の生物学的意義" Trends in Glycoscience and Glycotechnology. 2. (1990)

Yasuo Suzuki：“O__--乙酰化唾液酸的生物学意义”糖科学和糖技术趋势2。（1990）

Yasuo Suzuki,K.Sato,M.Kiso,A.Hasegawa: "New Ganglioside analogs that inhibite influenza virus sialidase" IXth International Catbohydrate Symposium(August 12ー17,1990,Yokohama,Japan),Abstract Book pp.240(1990). 240 (1990)

Yasuo Suzuki、K. Sato、M. Kiso、A. Hasekawa：“抑制流感病毒唾液酸酶的新神经节苷脂类似物”第九届国际猫碳水化合物研讨会（1990 年 8 月 12-17 日，日本横滨），摘要书第 240 页（1990 年） .240 (1990)

Yasuo Suzuki: "Glycolipid recognition of influenza virus hemagglutinin and neuraminidase" J.Glycosphingolipid Research. 1. 25-27 (1990)

Yasuo Suzuki：“流感病毒血凝素和神经氨酸酶的糖脂识别”J.Glycosphingolipid Research。

鈴木 康夫: "インフルエンザウイルスヘマグルチニンおよびノイラミュタンゼの細胞膜シアル酸含有糖鎖認識の機構" 生化学. 62. 231-260 (1990)

Yasuo Suzuki：“流感病毒血凝素和神经突变酶识别质膜唾液酸的糖链的机制”生物化学 62. 231-260 (1990)。