Foss Nicolai, Duranti Marcello, Magni Chiara, Frøkiaer Hanne
Biochemistry and Nutrition Group, BioCentrum-DTU, Technical University of Denmark, Kgs Lyngby, Denmark.
Int Arch Allergy Immunol. 2006;141(2):141-50. doi: 10.1159/000094716. Epub 2006 Jul 24.
The well-established murine model of IgE-mediated food allergy, based on oral administration of antigen and cholera toxin (CT), has within the previous years been used to evaluate various food proteins. Nonetheless, little knowledge on the factors that determine the allergenicity of food proteins is available so far. The use of proteins from the legume seed Lupinus albus as food ingredients calls for an evaluation of their allergenic potential, and therefore, we applied the cited model to investigate the putative allergenicity of three lupin protein preparations representing different matrices in which the four types of conglutins are present in varying concentrations.
Weekly, BALB/c A mice were orally immunized with the three lupin protein products together with CT. Total specific antibodies and IgE were determined by ELISA and Western blotting.
A dose-dependent Ig response against the analyzed proteins was observed for all three lupin products, while IgE responses against conglutins beta, gamma and delta, but not against conglutin alpha, were primarily detected after oral administration of lupin flakes. Whereas no differences among the samples for total specific Ig responses were seen, orally administered lupin flake extracts were much more efficient in inducing a conglutin-specific IgE response compared with fractionated lupin protein products.
Although the lupin-specific Ig response induced by coadministration of CT and lupin proteins appears to be dose dependent, the IgE response appears to depend merely on some intrinsic properties of the proteins as well as some factors of the protein matrix.
基于口服抗原和霍乱毒素(CT)建立的成熟的IgE介导的食物过敏小鼠模型,在过去几年中已被用于评估各种食物蛋白。然而,到目前为止,关于决定食物蛋白致敏性的因素知之甚少。将豆科植物种子白羽扇豆中的蛋白质用作食品成分需要评估其致敏潜力,因此,我们应用上述模型来研究三种羽扇豆蛋白制剂的假定致敏性,这三种制剂代表了不同的基质,其中四种凝集素以不同浓度存在。
每周给BALB/c A小鼠口服三种羽扇豆蛋白产品以及CT。通过ELISA和蛋白质印迹法测定总特异性抗体和IgE。
观察到所有三种羽扇豆产品对分析的蛋白质均有剂量依赖性的Ig反应,而口服羽扇豆片后主要检测到针对β、γ和δ凝集素的IgE反应,而未检测到针对α凝集素的反应。虽然在总特异性Ig反应的样本之间未观察到差异,但与分级分离的羽扇豆蛋白产品相比,口服羽扇豆片提取物在诱导凝集素特异性IgE反应方面效率更高。
虽然CT与羽扇豆蛋白共同给药诱导的羽扇豆特异性Ig反应似乎是剂量依赖性的,但IgE反应似乎仅取决于蛋白质的一些内在特性以及蛋白质基质的一些因素。