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Analysis of antigenic determinant sites of influenza virus using epitope scanning method

表位扫描法分析流感病毒抗原决定簇位点

基本信息

批准号：
05670287
负责人：
NAKAJIMA Setsuko
金额：
$ 1.34万
依托单位：
The Institute of Public Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670287/
关键词：
Multipin peptide synthesis Influenza virus HA protein Antigenic determinant site 抗原決定基 HAタンパク

项目摘要

The purpose of this study is to synthesize peptides continuously which cover antigenic region of the hemagglutinin (HA) protein of influenza virus, and determine the antigenic determinant sites of the HA protein recognized by the human serum antibody. Furtheremore, we will analyze the degree of immunogenicity of each antigenic determinant site. We have obtained the following results until now.1. Multipin peptide synthesis was done according to the amino acid sequence of the HA polypeptide of influenza virus A/Kamata/14/91 (H3N2). The peptides consisting of eight or ten amino acids were continuously synthesized skipping two amino acids, covering the amino acid seuences between 100 to 300 of the HA polypeptide. Five paired sera of the patients who were infected with influenza A (H3N2) during the 1990/91 influenza season were used. The identification and the degree of immunogenicity of the ocntinuous epitopes were analyzed by ELISA assay.2. Five sera obtained after postinfection had 4 to 32-fold increased HI titers compared to those of the sera obtained at early stage of infection. However, the latter sera except one had HI titers of 20 to 40. When peptide antigens consisting of eight amino acids were used, about 1/3 of the peptides reacted with paired sera at defferent degrees. In one case who had a low HI titer at earky stage of infectin, postinfection serum reacted specifically with a few peptides. However, we could not specify antigenic determinant sites. We further synthesized peptides consisting of 10 amino acids. This time, antigenic determinant sites became more specifically identified compared to those of 8 amino acids, but further experiments were necessary to get clearcut conclusions.3. Further analyzes with the selection of patient's sera, determination of the number of amino acids consisting of the peptides to get more specificity, or severe control to see the degree of antigenicity, and so on, remained to be done.

本研究的目的是连续合成覆盖流感病毒血凝素（HA）蛋白抗原区的多肽，并确定人血清抗体识别HA蛋白的抗原决定簇位点。进一步分析各抗原决定簇位点的免疫原性程度。到目前为止，我们已经得到了以下结果.根据流感病毒A/Kamata/14/91（H3 N2）的HA多肽的氨基酸序列进行多针肽合成。由8个或10个氨基酸组成的肽连续合成，跳过两个氨基酸，覆盖HA多肽的100至300之间的氨基酸序列。使用1990/91流感季节期间感染甲型流感（H3 N2）的患者的五对血清。采用ELISA法对所获得的连续表位进行鉴定，并分析其免疫原性.感染后获得的5份血清的HI滴度比感染早期获得的血清高4 ~ 32倍。然而，除一份外，后者血清的HI滴度为20至40。当使用由8个氨基酸组成的肽抗原时，约1/3的肽与配对血清有不同程度的反应。1例感染早期HI滴度低者，感染后血清与少数肽段特异性反应。然而，我们不能指定抗原决定簇位点。我们进一步合成了由10个氨基酸组成的肽。这一次，抗原决定簇位点变得比8个氨基酸的那些更特异性地鉴定，但需要进一步的实验来得到明确的结论。进一步的分析与患者的血清的选择，确定的氨基酸组成的肽的数量，以获得更多的特异性，或严格的控制，看看抗原性的程度，等等，仍然有待完成。