James J M, Sixbey J P, Helm R M, Bannon G A, Burks A W
Department of Pediatrics, University of Arkansas for Medical Sciences, Arkansas Children's Hospital Research Institute, Little Rock 72202, USA.
J Allergy Clin Immunol. 1997 Feb;99(2):239-44. doi: 10.1016/s0091-6749(97)70103-9.
Low molecular weight allergens may be responsible for hypersensitivity reactions after the ingestion of wheat.
The purpose of this investigation was to identify relevant, low molecular weight allergens after the ingestion of wheat protein.
Serum samples were collected from seven children with wheat allergy and one adult with baker's asthma. Control serum samples were collected from wheat-tolerant patients. Wheat extracts were prepared and separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in 12.5% gels revealing numerous protein bands. IgE immunoblot analysis of crude wheat extracts identified multiple IgE-binding proteins. Wheat proteins were separated further with two-dimensional gel electrophoresis, which was followed by IgE immunoblotting investigations.
Immunoblot analysis identified a 15 kd wheat protein that bound IgE from all five children with wheat allergy who were evaluated. No IgE binding to this wheat protein was demonstrated in any of the control subjects. Samples representing the 15 kd wheat protein (isoelective point, 5.85) were selected. The N-terminal peptide sequence of this protein (residues 1 to 20) matched to a wheat alpha-amylase inhibitor.
These data demonstrate that wheat alpha-amylase inhibitor is a relevant allergen in patients experiencing hypersensitivity reactions after the ingestion of wheat protein. This wheat protein, which has been implicated as an important allergen in patients with baker's asthma, represents a sensitizing allergen after both ingestion and inhalation.
低分子量变应原可能是摄入小麦后发生超敏反应的原因。
本研究的目的是鉴定摄入小麦蛋白后相关的低分子量变应原。
采集7名小麦过敏儿童和1名面包师哮喘成人的血清样本。从耐受小麦的患者中采集对照血清样本。制备小麦提取物,并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)在12.5%的凝胶中进行分离,显示出许多蛋白条带。对粗制小麦提取物进行IgE免疫印迹分析,鉴定出多种IgE结合蛋白。用二维凝胶电泳进一步分离小麦蛋白,随后进行IgE免疫印迹研究。
免疫印迹分析鉴定出一种15 kd的小麦蛋白,该蛋白与所有接受评估的5名小麦过敏儿童的IgE结合。在任何对照受试者中均未证明该小麦蛋白与IgE结合。选择代表15 kd小麦蛋白(等电点为5.85)的样本。该蛋白的N端肽序列(第1至20位氨基酸残基)与一种小麦α淀粉酶抑制剂匹配。
这些数据表明,小麦α淀粉酶抑制剂是摄入小麦蛋白后发生超敏反应患者的相关变应原。这种小麦蛋白在面包师哮喘患者中被认为是一种重要的变应原,在摄入和吸入后均代表一种致敏变应原。