虾过敏C57/BL6小鼠动物模型的建立

The establishment of C57/BL6 mice as a model of shrimp allergy

目的:建立C57/BL6小鼠虾过敏动物模型及其腹腔肥大细胞过敏模型。方法:运用虾蛋白粗提液免疫致敏C57/BL6小鼠,采用Western Blot鉴定致敏小鼠血清中特异性lgE和lgG1抗体;收集小鼠腹腔致敏肥大细胞(PMC),运用虾不同的过敏原组分诱导PMC体外定向释放组胺,HPLC和荧光酶标仪法检测组胺释放水平。结果:Western Blot结果显示致敏小鼠血清IgE和IgG1与相对分子量为36ku的原肌球蛋白反应率分别为57.1%和74.1%,与80ku过敏原的反应率均为42.9%,与21ku过敏原的反应率分别为42.9%和28.6%。HPLC和荧光酶标仪法检测PMC定向释放组胺的结果无显著差异,36ku的原肌球蛋白定向诱导组胺的释放率最高,分别为18.52%和21.59%,80ku和21ku蛋白次之,表明36ku的原肌球蛋白是C57/BL6小鼠的主要虾过敏原,21ku和80ku蛋白是次要过敏原,这与人类虾过敏的状况相一致。结论:C57/BL6小鼠是一种有效的虾过敏动物模型,其腹腔肥大细胞是一种可行的检测和评价食物过敏原的细胞模型。

In order to establish the C57/BL6 mouse allergic model and in vitro cell model of its mast cell, six-weekold C57/BL6 mice were immunized with shrimp protein extracts.Western Blot was used to analyze the specificity of sera IgE and lgG1 from the immunized mice.Sensitized peritoneal mast cells（PMC） were collected and induced to release histamine with various kinds of shrimp allergens, and then histamine was detected by HPLC and fluorescence microplate reader, respectively.Western Blot showed tropomyosin at 36ku could be recognized by 57.1% IgE and 74.1% lgG1 form immunized mice sera,80ku shrimp allergen by 42.9% IgGl and IgE,and 21ku shrimp allergen by 28.6% lgGl and 42.9 % lgE.The release rates of histamine detected by HPLC and fluorescence microplate reader with no significant difference, and the highest histamine release rate with 21.59% and 18.52% from PMC induced by tropomyosin, followed by 80ku and 21ku allergens. Tropomyosin was the major shrimp allergen of C57/BL6 mice,and 80ku and 21ku allergens were the minor allergens,which was identical with human shrimp allergy.The results demonstrated that C57/BL6 mouse was a very ideal shrimp allergic animal model and PMC was an available cell model to detect and assess food allergens.