Laccase/caffeic acid-catalyzed crosslinking coupled with galactomannan alters the conformational structure of ovalbumin and alleviates Th2-mediated allergic asthma
Ishfaq Ahmed , Suidong Ouyang , Shengquan Wu , Haochang Song , Miaoyuan Zhang , Renxing Luo , Peishan Lu , Jiaqi Deng , Tingting Zheng , Yanyan Wang , Xinguang Liu , Gonghua Huang
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Abstract
Ovalbumin (OVA) is the major allergenic protein that can induce T helper 2 (Th2)-allergic reactions, for which current treatment options are inadequate. In this study, we developed a polymerized hypoallergenic OVA product via laccase/caffeic acid (Lac/CA)-catalyzed crosslinking in conjunction with galactomannan (Man). The formation of high molecular weight crosslinked polymers and the IgG-binding were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The study indicated that Lac/CA-catalyzed crosslinking plus Man conjugation substantially altered secondary and tertiary structures of OVA along with the variation in surface hydrophobicity. Gastrointestinal digestion stability assay indicated that crosslinked OVA exhibited less resistance in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Mouse model study indicated that Lac-Man/OVA ameliorated eosinophilic airway inflammatory response and efficiently downregulated the expression of Th2-related cytokines (interleukin (IL)-4, IL-5, and IL-13), and upregulated IFN-γ and IL-10 expression. Stimulation of bone marrow-derived dendritic cells with Lac-Man/OVA suppressed the expression of phenotypic maturation markers (CD80 and CD86) and MHC class II molecules, and suppressed the expression levels of proinflammatory cytokines. The knowledge obtained in the present study offers an effective way to acquire a hypoallergenic OVA product that can have a therapeutic effect in alleviating OVA-induced allergic asthma.
卵清蛋白（OVA）是一种主要的致敏蛋白，可诱发T辅助细胞2（Th2）过敏反应，而目前的治疗方法还不足以解决这一问题。在这项研究中，我们通过漆酶/咖啡酸（Lac/CA）催化交联与半乳甘露聚糖（Man）结合，开发出了一种聚合的低过敏性 OVA 产品。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和 Western 印迹法分析了高分子量交联聚合物的形成和与 IgG 的结合。研究表明，Lac/CA 催化交联和 Man 共轭大大改变了 OVA 的二级和三级结构以及表面疏水性的变化。胃肠道消化稳定性试验表明，交联的 OVA 在模拟胃液（SGF）和模拟肠液（SIF）中表现出较低的抗性。小鼠模型研究表明，Lac-Man/OVA 可改善嗜酸性粒细胞气道炎症反应，有效下调 Th2 相关细胞因子（白细胞介素 (IL)-4、IL-5 和 IL-13）的表达，并上调 IFN-γ 和 IL-10 的表达。用 Lac-Man/OVA 刺激骨髓树突状细胞可抑制表型成熟标志物（CD80 和 CD86）和 MHC II 类分子的表达，并抑制促炎细胞因子的表达水平。本研究获得的知识为获得低过敏性 OVA 产品提供了一种有效途径，这种产品对缓解 OVA 引起的过敏性哮喘具有治疗作用。
期刊介绍：
Food Science and Human Wellness is an international peer-reviewed journal that provides a forum for the dissemination of the latest scientific results in food science, nutriology, immunology and cross-field research. Articles must present information that is novel, has high impact and interest, and is of high scientific quality. By their effort, it has been developed to promote the public awareness on diet, advocate healthy diet, reduce the harm caused by unreasonable dietary habit, and directs healthy food development for food industrial producers.
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