小麦主要致敏蛋白ω-5 醇溶蛋白HLA-DR分子限制性T细胞表位预测与鉴定

小麦依赖运动诱发严重过敏反应是食物过敏的特殊类型，患者在食入面食后4-6小时后运动即可发生严重过敏反应，ω-5醇溶蛋白是其发病的主要致敏蛋白。ω-5醇溶蛋白氨基酸序列已确定，并重组用于临床诊断。HLA-DRB1基因的高度多态性在一定程度上影响抗原提呈细胞提呈抗原肽种类以及激活特异性CD4+T细胞的能力，这可能是不同个体对同一蛋白组分产生不同免疫应答的原因。我们通过PCR-SBT方法对HLA-DRB1进行基因分型，分析出与ω-5醇溶蛋白致敏相关的HLA-DRB1等位基因；通过生物信息学方法利用T细胞表位预测网站预测出候选HLA-DR分子限制性T细胞表位，候选表位合成纯化后通过ELISPOT斑点酶联免疫分析技术、流式细胞术、CFSE标记技术对候选表位肽进行功能验证，从而鉴定出优势T细胞表位。ω-5醇溶蛋白HLA-DR限制性T细胞表位鉴定为变应原特异性免疫治疗提供了新的靶点。

Wheat dependent exercise induced anaphylaxis (WDEIA)is a special type of food allergy，patiens will experience severse allergic reaction when they exercise after 4-6 hours eating wheat food. ω-5 gliadin is the major allergen causing WDEIA ,which has been recombined and used for the diagnosis of WDEIA .To a certain extent,the high polymorphisms of HLA-DRB1 gene influence the ability of presentation antigen ,which cause the diffence of T cell response to same allergen component among individuals. In this research,we collect the subjects of WDEIA and the healthy individuals for HLA-DRB1 genotyping by PCR-SBT.We analyze the HLA-DRB1 genotype that is associated with ω-5 gliadin sensitization.The prediction of HLA-DR restricted T cell epitope derived from the ω-5 gliadin is based on the web of Syfpeithy 、MHC-THREAD、 Epipredict、ProPred. We characterized synthetic candidate epitopes in relative HLA -DRB1 genotype positive healthy individuals by using Elispot assay. We evaluate the ability of proliferation of the epitope-specific CD4+ T cell by CFSE staining. Prediction and identification of HLA-DR restricted T cell epitope derived from ω-5 gliadin which is utility in epitope design for the development of HLA based vaccines and antigen-specific immunotherapy.