食品过敏原体外激发小鼠致敏肥大细胞组胺释放  

Histamine releasing from sensitized Mast cells from mouse model stimu la ted by food allergen in vitro

作　　者：向军俭[1] 张在军[1] 毛露甜[2] 邓宁[1] 杨红宇[1] 

机构地区：[1]暨南大学生命科学技术学院分子免疫与抗体工程实验室,广州510630 [2]广东省惠州学院生物系,广东惠州516015

出　　处：《广东医学》2005年第5期593-595,共3页Guangdong Medical Journal

基　　金：广东省自然科学基金资助项目[编号:2003(36708)]

摘　　要：目的检测食品过敏原体外激发小鼠致敏肥大细胞的组胺释放率,探讨食品过敏原鉴定与评价方法。方法以河虾蛋白和鸡卵清蛋白,氢氧化铝佐剂免疫Balb/c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7,14,21天血清中过敏原sIgE水平;并在激发后第14天分离小鼠腹腔肥大细胞,过敏原体外激发,荧光光度法检测组胺释放量,计算释放率,Student’s统计分析显著性意义。结果①模型组小鼠血清sIgE水平比对照组明显升高,且在过敏原激发后第14天sIgE效价最高,河虾和鸡卵清蛋白组分别达到1/6400和1/3200。②鸡卵清蛋白组定向组胺释放率达60.75%;河虾组腹腔或皮下免疫分别达到71.53%和88.48%,所有模型组与对照组比较均呈显著差异(P<0.05)。结论利用小鼠过敏模型,检测食品过敏原体外激发组胺释放率可能成为食品过敏原鉴定与评价的方法。Objective To develop a novel te c hnique based on determining histamine release from mouse sensitized cells stimul ated in vitro for assessing and identifying food allergens. Methods The Balb/c mice were immunized by injection of ri ver shrimp proteins or HEW(hen egg white) 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、 14 and 21 days after challenge; and the peritoneal Mast cells were collecte d at 14th day and were stimulated by allergen in vitro. The liberation of histamine was measured by fluorescence assay and was converted to releasing rate for statistical analysis with Student's system. Results ①The levels of sIgE in the experimental groups w ere higher than that in control at three different time points. They reached the peak at 14 day after challenge, with a value of 1/6?400 in riv er shrimp and 1/3?200 in HEW group. ② The histamine directional liberation ratio group was 60.75% in HEW group and 88.48% was 37.19% in control group; The liberation ratiowas 71.53% and by intraperitoneal injection (i.p) or 88. 48% by hypodermic injection (i.h), but it was was 43.76% in the control. All the model groups revealed significant difference compared with the control(P <0.05). Conclusion The present data indicated that histamine release test with sensitized cells fro m mouse model challenged by food allergen in vitro maybe a prospective technique for assessing and identifying food allergens.

关 键 词：过敏原 激发 食品 体外 组胺释放 致敏 间接ELISA测定 Balb/c小鼠 IGE水平 氢氧化铝佐剂 腹腔肥大细胞 荧光光度法 释放率 卵清蛋白 动物模型 食物过敏 高反应性 统计分析 小鼠血清 sIgE 蛋白组分 皮下免疫 对照组 检测 

分 类 号：R725.622.5[医药卫生—儿科] R155.5[医药卫生—临床医学]