Forty-two patients with WDEIA (5 negative/37 positive for ω-5 gliadin sIgE) were enrolled. For comparison, 8 patients with immediate-type wheat allergy without WDEIA and 20 healthy controls without wheat allergy were also enrolled. Extracted wheat proteins were separated by 2D-PAGE. Proteins that reacted with serum IgE antibody in 2D Western blotting (2D-WB) were identified using mass spectrometry. Recombinant proteins were synthesized in Escherichia coli, and the antigenicity was tested using ELISA and the basophil activation test.
In 2D-WB, nine proteins reacted with the serum IgE antibody from at least 60% of patients with WDEIA (n ≥ 25/42). ELISA revealed that alpha/beta gliadin MM1 exhibited the highest positive immunoreactivity in 23 of 26 patients who were positive for ω-5 gliadin sIgE (88%) and in 5 of 5 patients who were negative for ω-5 gliadin sIgE (100%). Alpha/beta gliadin MM1 exhibited significantly higher basophil activation in 14 patients with WDEIA when compared to 5 individuals without a wheat allergy.
Alpha/beta gliadin MM1 sIgE exhibited the highest seropositivity, even among patients who were negative for ω-5 gliadin sIgE. The inclusion of alpha/beta gliadin MM1 in allergen-sIgE tests may improve the sensitivity for diagnosing WDEIA.
方法:纳入42例WDEIA患者(ω-5麦醇溶蛋白sIgE阴性/阳性37例)。为了比较,还招募了8名没有WDEIA的立即型小麦过敏患者和20名没有小麦过敏的健康对照。通过2D-PAGE分离提取的小麦蛋白。使用质谱鉴定在2DWestern印迹(2D-WB)中与血清IgE抗体反应的蛋白质。重组蛋白在大肠杆菌中合成,用ELISA和嗜碱性粒细胞活化试验检测抗原性。
结果:在2D-WB中,来自至少60%的WDEIA患者(n≥25/42)的9种蛋白质与血清IgE抗体发生反应.ELISA显示,α/β麦醇溶蛋白MM1在ω-5麦醇溶蛋白sIgE阳性的26例患者中有23例(88%)和ω-5麦醇溶蛋白sIgE阴性的5例患者中表现出最高的阳性免疫反应性(100%)。与5名没有小麦过敏的个体相比,14名WDEIA患者的α/β麦醇溶蛋白MM1表现出明显更高的嗜碱性粒细胞活化。
结论:α/β麦醇溶蛋白MM1sIgE表现出最高的血清阳性,甚至在ω-5麦醇溶蛋白sIgE阴性的患者中。在过敏原-sIgE测试中包含α/β麦醇溶蛋白MM1可能会提高诊断WDEIA的敏感性。