文献类型：期刊文献

中文题名：具有T细胞口服免疫耐受作用的β-乳球蛋白水解物的制备及鉴定

英文题名：Preparation and Identification of β-Lactoglobulin Hydrolysates with T-Cell Oral Immunotolerance

作者：田岭含[1];张倩倩[1];丛艳君[1];闫文杰[2]

第一作者：田岭含

机构：[1]北京工商大学食品与健康学院/北京市食品营养与人类健康高精尖创新中心/北京市食品添加剂工程技术研究中心,北京100048;[2]北京联合大学生物化学工程学院,北京100023

第一机构：北京工商大学食品与健康学院/北京市食品营养与人类健康高精尖创新中心/北京市食品添加剂工程技术研究中心,北京100048

年份：2023

卷号：41

期号：2

起止页码：57-69

中文期刊名：食品科学技术学报

外文期刊名：Journal of Food Science and Technology

收录：CSTPCD;;EI(收录号：20231914080023);Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家重点研发计划项目子课题(2019YFC1605002);国家自然科学基金资助项目(31872886)。

语种：中文

中文关键词：牛乳过敏;β-乳球蛋白;多肽;口服耐受;T细胞表位

外文关键词：cow's milk allergy;β-lactoglobulin;polypeptide;oral tolerance;T cell epitope

摘要：以牛乳中主要过敏原β-乳球蛋白为研究对象,制备和鉴定具有T细胞表位和口服免疫耐受性的水解物,旨在为牛乳过敏患者口服免疫治疗方法提供理论依据。采用生物信息学方法预测β-乳球蛋白的T细胞表位,通过质谱分析6种蛋白酶水解β-乳球蛋白水解物的氨基酸序列,用T细胞增殖实验鉴定水解物中多肽免疫耐受作用,体内实验测定小鼠血清特异性抗体(IgE、IgG1、IgG2a)、Th1细胞因子(IFN-γ、IL-17)、Th2细胞因子(IL-4、IL-5、IL-13)、组胺、几丁质酶-3样蛋白1的水平及小鼠脾脏细胞亚群的变化水平,验证β-乳球蛋白水解物的口服免疫治疗活性。研究结果表明:胰蛋白酶、复合蛋白酶和木瓜蛋白酶水解物具有口服耐受性,其中复合蛋白酶和木瓜蛋白酶水解物具有口服耐受性是创新性发现。中性蛋白酶水解物虽然含有T细胞表位,但是仍然具有致敏性。因此含有T细胞表位的水解物不一定具有口服免疫耐受性,需要体内实验验证。研究结论以期为新型抗过敏乳基料的开发和临床治疗牛乳过敏提供参考数据。
β-Lactoglobulin,one of major allergens in cow milk,was studied in this paper,and the T cell immune tolerance hydrolysates ofβ-lactoglobulin were prepared and identified to provide a theoretical basis for oral immunotherapy for patients with cow milk allergy.Firstly,the bioinformatics method was used to predict the possibleβ-lactoglobulin T-cell epitopes,and then,β-lactoglobulin was respectively hydrolyzed by six kinds of protease.The amino acid sequences of polypeptide ofβ-lactoglobulin hydrolysates were analyzed by mass spectrometry,and these peptides were synthesized by solid phase synthesis.The immune tolerance of the peptides were identified by T cell proliferation test.Therefore,trypsin,protamex,papain and neutrase hydrolysates were initially screened for T cell immune tolerance.Furthermore,the immune tolerance ofβ-lactoglobulin hydrolysates were evaluated by animal experiments.The levels of specific antibody(IgE,IgG 1,IgG 2a),Th1 cytokine(IFN-γ,IL-17),Th2 cytokine(IL-4,IL-5,IL-13),histamine,chitinase-3-like protein 1 in mice serum and the differentiation of mice spleen cell subsets were measured in vivo.The results showed that trypsin,protamex and papain hydrolysates had oral tolerance,among which protamex and papain hydrolysates had been reported innovatively.Although neutrase protease hydrolysates contained T cell epitopes,they were still allergenic.Therefore,the hydrolysates containing T cell epitopes did not necessarily have oral immune tolerance and need to be verified in vivo.The conclusion of this study could provide an important theoretical basis for the development of new anti-allergy milk-based materials and clinical treatment of milk allergy.