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Reduction of antigenicity of allergen proteins by the attachment of polysaccjarides and induction of immune tolerance

通过附着多糖降低过敏原蛋白的抗原性并诱导免疫耐受

基本信息

批准号：
13556019
负责人：
KATO Akio
金额：
$ 6.02万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13556019/
关键词：
Lysozyme Japanese ceder pollen allergen Cryj 1 soy allergen protein-polysaccharicle conjugate Immune tolerance S.cerevisiae Reduction of antigen city スギアレルゲン経口免疫寛容リゾチーム-抗体 IgE IgG アレルギースギ花粉症 Cryj1 多糖修飾メイラード型蛋白-多糖複合体

项目摘要

An attempt was made to mask the allergenic structure of major allergen protein, Cryj 1., in Japanese cedar pollen using the Maillard-type polysaccharide conjugation. The SDS-PAGE pattern of the Cryj 1-galactomannan conjugate prepared by Maillard reaction showed the broad bands widely distributed from 50 kDa to more than 100 kDa, suggesting the attachment of galactomannan. The competitive enzyme linked immunosorbent assay revealed that the IgE antibody in the sera of ceder pollen-sensitive patients strongly reacted with Cryj 1, while it did not react with the Cryj 1-galactomannan conjugate. This result suggests that the allergenicity of Cryj 1 is greatly reduced by the conjugation with galactomannan. The soy protein-chitosan conjugate was also formed by the Maillard reaction in dry state (relative humidity 65%) at 60 ℃ for 2 weeks to improve the functional properties. The antimicrobial activity of the Maillard-type soy protein-chitosan conjugates enhanced 2〜3 times that of soy protein-chitosan mixture. The soy protein-chitosan conjugate showed excellent emulsifying property with the progress of Maillard-type conjugation. The allergenicity of soy protein was greatly decreased by the attachment of chitosan through Maillard reaction. The immonoblotting analysis with patient's sera revealed that soy protein-chitosan conjugate was more effective to mask the allergen structure of soy protein causing from 34kDa-protein (Gly m Bd 30K) than soy protein-galactomannan conjugate. The western blotting showed that allergen (34kDa-protein) was completely masked by soy protein-chitosan conjugation, while it was not completely masked by soy protein galactomannan conjugation

试图掩盖主要过敏原蛋白Cryj 1的过敏原结构，在日本雪松花粉使用美拉德型多糖共轭。通过Maillard反应制备的Cryj 1-半乳甘露聚糖偶联物的SDS-PAGE图谱显示宽带分布在50 kDa至100 kDa以上，表明半乳甘露聚糖的附着。竞争性酶联免疫吸附试验表明，雪松花粉敏感患者血清中的IgE抗体与Cryj 1强烈反应，而它不与Cryj 1-半乳甘露聚糖结合物反应。该结果表明，通过与半乳甘露聚糖缀合，Cryj 1的变应原性大大降低。在干燥状态下（相对湿度65%），于60 ℃下反应2周，制备大豆蛋白-壳聚糖结合物，以改善其功能特性。美拉德型大豆蛋白-壳聚糖偶联物的抗菌活性是大豆蛋白-壳聚糖混合物的2 ~ 3倍。大豆蛋白-壳聚糖复合物具有良好的乳化性能，且随着Maillard型复合的发展而不断提高。通过美拉德反应将壳聚糖接枝到大豆蛋白上，大大降低了大豆蛋白的致敏性。用患者血清进行的免疫印迹分析显示，大豆蛋白-壳聚糖缀合物比大豆蛋白-半乳甘露聚糖缀合物更有效地掩蔽由34 kDa蛋白（Gly m Bd 30 K）引起的大豆蛋白过敏原结构。免疫印迹结果表明，大豆蛋白-壳聚糖结合物可完全掩蔽34 kDa蛋白，而大豆蛋白-半乳甘露聚糖结合物不能完全掩蔽34 kDa蛋白