Food Allergy & Immunology Laboratory, Department of Food Science & Human Nutrition, Michigan State University, East Lansing, MI 48824, USA.
Wheat Breeding & Genetics Laboratory, Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI 48824, USA.
Int J Mol Sci. 2022 Jun 10;23(12):6505. doi: 10.3390/ijms23126505.
Wheat allergies are potentially life-threatening because of the high risk of anaphylaxis. Wheats belong to four genotypes represented in thousands of lines and varieties. Monitoring changes to wheat allergens is critical to prevent inadvertent ntroduction of hyper-allergenic varieties via breeding. However, validated methods for this purpose are unavailable at present. As a proof-of-concept study, we tested the hypothesis that salt-soluble wheat allergens in our mouse model will be identical to those reported for humans. Groups of Balb/cJ mice were rendered allergic to durum wheat salt-soluble protein extract (SSPE). Using blood from allergic mice, a mini hyper-IgE plasma bank was created and used in optimizing an IgE Western blotting (IEWB) to identify IgE binding allergens. The LC-MS/MS was used to sequence the allergenic bands. An ancient wheat was grown in our greenhouse and extracted SSPE. Using the optimized IEWB method followed by sequencing, the cross-reacting allergens in wheat were identified. Database analysis showed all but 2 of the durum wheat allergens and all wheat allergens identified in this model had been reported as human allergens. Thus, this model may be used to identify and monitor potential changes to salt-soluble wheat allergens caused by breeding.
小麦过敏症具有潜在的生命威胁，因为其存在严重过敏反应的高风险。小麦属于四种基因型，有数千种品系和品种。监测小麦过敏原的变化对于防止通过育种无意中引入高致敏性品种至关重要。然而，目前没有为此目的验证的方法。作为概念验证研究，我们测试了这样一个假设，即在我们的小鼠模型中，盐溶性小麦过敏原将与报告的人类过敏原相同。将 Balb/cJ 组小鼠致敏于硬质小麦盐溶性蛋白提取物（SSPE）。使用来自过敏小鼠的血液，创建了一个小型超 IgE 血浆库，并用于优化 IgE 免疫印迹（IEWB）以鉴定 IgE 结合过敏原。LC-MS/MS 用于对过敏原条带进行测序。在我们的温室中种植了一种古老的小麦并提取了 SSPE。使用优化的 IEWB 方法和测序，鉴定了小麦中的交叉反应过敏原。数据库分析显示，在本模型中鉴定出的除 2 种硬质小麦过敏原和所有小麦过敏原均被报告为人类过敏原。因此，该模型可用于鉴定和监测由于育种而导致的盐溶性小麦过敏原的潜在变化。
