摘要
建立牛乳蛋白、河虾蛋白的ICR小鼠过敏模型,检测食品过敏原体外激发小鼠致肥大细胞的组胺释放率,探讨食品过敏原鉴定并验证所建过敏原体外评价新方法。方法以牛奶蛋白、河虾蛋白,氢氧化铝佐剂免疫ICR小鼠,建立IgE高反应性食物过敏动物模型。间接ELISA测定激发后第7、14、21d血清中过敏原sIgE水平;并在激发第14d分离小鼠腹腔肥大细胞,过敏原体外激发,紫外分光光度法检测类胰蛋白酶定向释放水平。①模型小鼠血清sIgE水平明显高于对照组,且在过敏激发后第14dsIgE效价最高,河虾组和牛奶蛋白组的sIgE效价分别达到1/25 600和1/12 800。②牛奶组类胰蛋白酶释放率达38.5%;河虾组的类胰蛋白酶释放率达47.1%,所有模型组与对照组间差异均达显著水平(P<0.05)。建立了一种小鼠过敏模型,通过检测过敏原体外激发类胰蛋白酶释放率可以作为一种鉴定与评价食品过敏原的有效方法。
Objectives To establish allergic model with ICR mice for milk and shrimp protein,develop a novel technique based on determining histamine release from mouse sensitized cells stimulated in vitro for assessing and identifying food allergens.Methods The ICR mice were immunized by injection of river shrimp proteins or milk proteins with aluminum hydroxide as adjuvant.The sensitized mouse were challenged for one time at 7th day after sensitization.The sIgE of food allergens was measured by indirect ELISA at 7/14and 21days after challenge;and the peritoneal Mast cells were collected at 14th day and were stimulated by allergen in vitro.The liberation of tryptase was measured by ultraviolet spectrophotometry assay and was converted to releasing rate for statistical analysis with Student's system.Results ①The levels of sIgE in the experimental groups were higher than on control at three different points.They reached the peak at14day after challenge,with a value of 1/25600 in river shrimp and 1/12800 in milk group.②The tryptase liberation ratio group was 38.5% in milk group and 36.14% in control group;The liberation ratio was 74.5% in river shrimp group and 47.1% in the control.All the model groups revealed significant difference compared with the control(P< 0.05).Conclusion The present data indicated that tryptase release test with sensitized cells from mouse model challenged by food allergen in vitro maybe a prospective technique for assessing and identifying food allergens.
出处
《食品与生物技术学报》
CAS
CSCD
北大核心
2012年第5期499-504,共6页
Journal of Food Science and Biotechnology
基金
国家自然科学基金项目(20806033)
国家“十二五”科技支撑计划项目(2011BAK10B03)
国家973计划项目(2012CB720804)
2010年度公益性行业(农业)科研专项项目(201003008-08)
江苏省质监局立项项目(KJ122704)
