摘要
小麦是八大类食物致敏原之一,近期研究表明,球蛋白1S(globulin-1 S allele,GSA)是一种糖基化小麦致敏蛋白。为解析GSA致敏机制并探究糖基化在GSA致敏性中所发挥的作用,本研究利用N-糖肽酶F(peptide-N-glycosidase F,PNGase F)去除GSA上的N-糖基化,并建立DC2.4树突状细胞模型探究糖基化对GSA免疫识别的影响。蛋白荧光标记法揭示了糖基化对DC2.4免疫识别GSA的促进作用,荧光定量PCR实验结果发现,糖基化GSA激活DC2.4表达细胞因子肿瘤坏死因子a(tumour necrosis factor α,TNF-α)。利用siRNA在DC2.4中对潜在的糖基化受体进行基因敲减,在此基础上挖掘糖基化GSA受体及其功能,结果显示C型凝集素(C-type lectin receptors,CLRs)受体树突状细胞特异性细胞间黏附分子3捕获非整联蛋白(dendritic cell-specific interellular adhesion molecule-3-grabbing non-integrin,DC-SIGN)介导了GSA的免疫激活活性。本研究发现DC-SIGN介导小麦糖基化致敏原球蛋白1S免疫识别并初步解析其机理,为小麦过敏症的预防和控制提供了潜在的靶点。
Wheat is one of the major food allergen sources worldwide.Recent studies identified globulin-1 S allele(GSA)as a novel glycosylated allergen in wheat.To elucidate the mechanism of GSA-induced allergy and investigate the role of glycosylation,we used PNGase F to eliminate N-glycans on GSA and established a DC2.4 dendritic cell model to study the role of glycosylation in the immune recognition of GSA.By using fluorescent-labeled GSA,we proved that glycosylation promoted the immune recognition of GSA by DC2.4.Further qPCR experiments revealed that glycosylation mediated the expression of cytokine TNF-α in DC2.4 after GSA stimulation.Moreover,the potential glycan receptors were knocked out by siRNA in DC2.4,thus elucidating the specific receptor of glycosylated GSA and the relevant function.Our results demonstrate that the C-type lectin receptor DC SIGN mediated the immune stimulation activity of GSA.This work elucidates the mechanism of DC SIGN mediated immune recognition of wheat allergen GSA,provides potential targets for the prevention and control of wheat allergy.
作者
王翀
朱城波
王彦波
傅玲琳
WANG Chong;ZHU Chengbo;WANG Yanbo;FU Linglin(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)
出处
《未来食品科学》
2021年第4期1-10,共10页
Future Food Science
基金
“十三五”国家重点研发计划项目(2019YFC1605002)。
