核桃中主要过敏原Jug r 1的致敏性研究
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摘要
近些年来,食物过敏的发病率呈逐年上升趋势。核桃是引起食物过敏的主要食品之一。目的:本文旨在研究核桃中主要过敏原Jug r 1与致敏性相关的免疫学特征,确定其线性抗原表位。方法:首先对核桃过敏原Jug r 1进行消化稳定性和热稳定性的研究,然后利用人体血清学分析其特异性抗体的结合情况,结合生物信息学对其氨基酸一级序列的相关特征参数的分析,预测其可能存在线性抗原表位,同时合成覆盖其全部氨基酸序列的重叠肽库,利用圆点免疫法(dot-blot)筛选出可与过敏患者血清中Ig E特异性结合的肽段,从而确定出Jug r1的线性抗原表位。结果 :核桃中主要过敏原Jug r 1在模拟胃液中60 min时仍未被完全消化;在90℃水浴中加热60 min其条带仍然存在,且可以与核桃过敏患者血清中的Ig E特异性结合。通过生物信息学软件和血清筛选出可结合的抗原表位。结论:核桃中主要过敏原Jug r 1具有较强的免疫学特性,且其存在3个免疫显性的抗原表位。对深入研究其过敏原改造和过敏机理提供理论依据。
The annual rate of food allergy increased gradually. As a widely consumed nut, walnut is an important kind of allergen. Objective: This study was aimed at immunity characteristics of the Jug r 1 which is the major allergen of walnut, and mapping the linear epitopes. Method: The digestion resistibility and the heat stability of Jug r 1 were estimated by simulated gastric digestion and high-temperature processing essay. The capacity of binding specific Ig E was compared with human serological analysis. The potential epitopes could be predicted using the parameter information DNAstar. And then overlapping peptides were synthesized according to the primary sequence of the amino acids. The linear epitopes were mapped by Dot-blot. Result: The major allergen of walnut, Jug r 1, was stable during the heating and digestion. Also Jug r 1 could bind the Ig E from patients' serum. The linear epitopes of Jug r 1 were mapped using the bioinformatic tools and Dot-blot. Conclusion: The immunity characteristics of Jug r 1 were stable and there were 3 linear eptiopes. This research could provide a reference for the allergen modification and allergic mechanism research.
The annual rate of food allergy increased gradually. As a widely consumed nut, walnut is an important kind of allergen. Objective: This study was aimed at immunity characteristics of the Jug r 1 which is the major allergen of walnut, and mapping the linear epitopes. Method: The digestion resistibility and the heat stability of Jug r 1 were estimated by simulated gastric digestion and high-temperature processing essay. The capacity of binding specific Ig E was compared with human serological analysis. The potential epitopes could be predicted using the parameter information DNAstar. And then overlapping peptides were synthesized according to the primary sequence of the amino acids. The linear epitopes were mapped by Dot-blot. Result: The major allergen of walnut, Jug r 1, was stable during the heating and digestion. Also Jug r 1 could bind the Ig E from patients' serum. The linear epitopes of Jug r 1 were mapped using the bioinformatic tools and Dot-blot. Conclusion: The immunity characteristics of Jug r 1 were stable and there were 3 linear eptiopes. This research could provide a reference for the allergen modification and allergic mechanism research.
引文
[1]刘玲,韩本勇,陈朝银.核桃蛋白研究进展[J].食品与发酵工业,2009,35(9):116-118.
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[6]方娟,陈朝银,赵声兰,等.核桃过敏原及其检测方法的研究进展[J].食品工业科技,2013,34(23):361-365.
[7]PASTORELLO E A,FARIOLIL L,PRAVETTONI V,et al.Lipid transfer protein and vicilin are important walnut allergens in patients not allergic to pollen[J].Journal of Allergy and Clinical Immunology,2004,114(4):908-914.
[8]ALLERGENS T N.Tree nut allergens[J].International Archives of Allergy and Immunology,2003,131(4):234-244.
[9]LWW P W,HEFLE S L,TAYLOR S L.Sandwich enzyme-linked immunosorbent assay(ELISA)for detection of mustard in foods[J].Journal of Food Science,2008,73(4):62-68.
[10]WEGRZYN N K,SAMPSON H A.Adverse reactions to foods[J].The Medical Clinics of North America,2006,90(1):97-127.
[11]BENDER A E,MATTHEWS D R.Adverse reactions to foods[J].British Journal of Nutrition,1981,46(1):403-407.
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[13]BARRE A,BORGES J P,CULERRIER R,et al.Homology modelling of the major peanut allergen Ara h 2 and surface mapping of Ig E-binding epitopes[J].Immunology Letters,2005,100(2):153-158.
[14]ALVAREZ P A,BOYE J I.Food production and processing considerations of allergenic food ingredients:a review[J].Journal of Allergy,2012,2012:746125.
[15]TEUBER S S,DANDEKAR A M,PETERSON W R,et al.Cloning and sequencing of a gene encoding a 2S albumin seed storage protein precursor from English walnut(Juglans regia),a major food allergen[J].Journal of Allergy and Clinical Immunology,1998,101(6):807-814.
[16]SORDET C,CULERRIER R,GRANIER C,et al.Expression of Jug r 1,the 2S albumin allergen from walnut(Juglans regia),as a correctly folded and functional recombinant protein[J].Peptides,2009,30(7):1213-1221.
[17]ROBOTHAM J M,TEUBER S S,SATHE S K,et al.Linear Ig E epitope mapping of the English walnut(Juglans regia)major food allergen,Jug r 1[J].Journal of Allergy and Clinical Immunology,2002,109(1):143-149.
[18]ASTWOOD J D,LEACH J N,FUCHS R L.Stability of food allergens to digestion in vitro[J].Nat Biotechnol,1996,14(10):1269-1273.
[19]蒋显斌,樊庆鲁,黄芊,等.抗除草剂转基因稻米蛋白的消化稳定性和热稳定性分析[J].西南农业学报,2014,27(5):1797-1800.
[20]刘光明,沈苑,曹敏杰,等.虾类过敏原的识别、纯化和检测技术研究[J].中国食品学报,2008,8(6):142-148.
[21]ZHENG L N,HONG L,PAWAR R,et al.Mapping Ig E binding epitopes of major shrimp(Penaeus monodon)al lergen with immunoinformatics tools[J].Food&Chemical Toxicology An International Journal Published for the British Industrial Biological Research Association,2011,49(11):2954-2960.
[22]SORDET C,CULERRIER R,GRANIER C,et al.Expression of Jug r 1,the 2S albumin allergen from walnut(Juglans regia),as a correctly folded and functional recombinant protein[J].Peptides,2009,30(7):1213-1221.
[23]蔡秋凤,王锡昌,郭城,等.加工处理对鲢小清蛋白免疫活性的影响[J].水产学报,2013,37(2):297-302.
[24]GROZDANOVIC M,POPOVIC M,POLOVIC N,et al.Evaluation of Ig E reactivity of active and thermally inactivated actinidin,a biomarker of kiwifruit allergy[J].Food and Chemical Toxicology,2012,50(3/4):1013-1018.
[25]郑礼娜,林洪,刘一璇,等.不同热加工方式对刀额新对虾过敏原活性的影响[J].水产学报,2011,35(2):466-471.
[26]TONG P,GAO J Y,CHEN H B,et al.Effect of heat treatment on the potential allergenicity and conformational structure of egg allergen ovotransferrin[J].Food Chemistry,2012,131(2):603-610.
[27]CABANILLAS B,MALEKI S J,RODR魱GUEZ J,et al.Heat and pressure treatments effects on peanut allergenicity[J].Food Chemistry,2012,132(1):360-366.
[28]KORBER B,LABUTE M,YUSIM K.Immunoinformatics comes of age[J].Plos Computational Biology,2006,2(6):e71.
[29]SAHA S,RAGHAVA G P.Prediction of continuous B-cell epitopes in an antigen using recurrent neural network[J].Proteins,2006,65(1):4048.
[30]夏宏林,何颖,邹泽红,等.苹果过敏原Mal d4蛋白抗原表位预测及交叉反应分析[J].中国免疫学杂志,2012(1):57-61.
[31]李江英,白雪娟,梁艳,等.用DNAStar软件预测Rv1410c结核分枝杆菌蛋白抗原表位[J].细胞与分子免疫学杂志,2015,31(4):474-477.
[32]李海侠,毛旭虎.蛋白质抗原表位研究进展[J].微生物学免疫学进展,2007,35(1):54-58.
[33]LEE J M,KIM E J,KWON D G,et al.Clinical characteristics of walnut allergy and evaluation of cross-reactivity between walnut and peanut in children under 4 years of age[J].Pediatric Allergy&Respiratory Disease,2011,21(4):261-268.
[2]NIEMANN L,TAYLOR S L,HEFLE S L.Detection of walnut residues in foods using an enzyme-linked immunosorbent assay[J].Journal of Food Science,2009,74(6):51-57.
[3]SAKAI S,ADACHI R,AKIYAMA H,et al.Determination of walnut protein in processed foods by enzyme-linked immunosorbent assay:interlaboratory study[J].Food Composition and Additives,2010,93(4):1255-1261.
[4]SICHERER S H,FURLONG A M,SAMPSON H A.Prevalence of peanut and tree nut allergy in the United States determined by means of a random digit dial telephone survey:a 5-year follow-up study[J].The Journal of Allergy and Clinical Immunology.2003,112(6):1203-1207.
[5]WANG H Y,YUAN F,WU Y J,et al.Detection of allergen walnut component in food by an improved RealTime PCR method[J].Journal of Food Protection,2009,72(11):2433-2435.
[6]方娟,陈朝银,赵声兰,等.核桃过敏原及其检测方法的研究进展[J].食品工业科技,2013,34(23):361-365.
[7]PASTORELLO E A,FARIOLIL L,PRAVETTONI V,et al.Lipid transfer protein and vicilin are important walnut allergens in patients not allergic to pollen[J].Journal of Allergy and Clinical Immunology,2004,114(4):908-914.
[8]ALLERGENS T N.Tree nut allergens[J].International Archives of Allergy and Immunology,2003,131(4):234-244.
[9]LWW P W,HEFLE S L,TAYLOR S L.Sandwich enzyme-linked immunosorbent assay(ELISA)for detection of mustard in foods[J].Journal of Food Science,2008,73(4):62-68.
[10]WEGRZYN N K,SAMPSON H A.Adverse reactions to foods[J].The Medical Clinics of North America,2006,90(1):97-127.
[11]BENDER A E,MATTHEWS D R.Adverse reactions to foods[J].British Journal of Nutrition,1981,46(1):403-407.
[12]BRUIJNZEEL K C,ORTOLANI C,AAS K,et al.Adverse reactions to food[J].Allergy,1995,51(8):623-635.
[13]BARRE A,BORGES J P,CULERRIER R,et al.Homology modelling of the major peanut allergen Ara h 2 and surface mapping of Ig E-binding epitopes[J].Immunology Letters,2005,100(2):153-158.
[14]ALVAREZ P A,BOYE J I.Food production and processing considerations of allergenic food ingredients:a review[J].Journal of Allergy,2012,2012:746125.
[15]TEUBER S S,DANDEKAR A M,PETERSON W R,et al.Cloning and sequencing of a gene encoding a 2S albumin seed storage protein precursor from English walnut(Juglans regia),a major food allergen[J].Journal of Allergy and Clinical Immunology,1998,101(6):807-814.
[16]SORDET C,CULERRIER R,GRANIER C,et al.Expression of Jug r 1,the 2S albumin allergen from walnut(Juglans regia),as a correctly folded and functional recombinant protein[J].Peptides,2009,30(7):1213-1221.
[17]ROBOTHAM J M,TEUBER S S,SATHE S K,et al.Linear Ig E epitope mapping of the English walnut(Juglans regia)major food allergen,Jug r 1[J].Journal of Allergy and Clinical Immunology,2002,109(1):143-149.
[18]ASTWOOD J D,LEACH J N,FUCHS R L.Stability of food allergens to digestion in vitro[J].Nat Biotechnol,1996,14(10):1269-1273.
[19]蒋显斌,樊庆鲁,黄芊,等.抗除草剂转基因稻米蛋白的消化稳定性和热稳定性分析[J].西南农业学报,2014,27(5):1797-1800.
[20]刘光明,沈苑,曹敏杰,等.虾类过敏原的识别、纯化和检测技术研究[J].中国食品学报,2008,8(6):142-148.
[21]ZHENG L N,HONG L,PAWAR R,et al.Mapping Ig E binding epitopes of major shrimp(Penaeus monodon)al lergen with immunoinformatics tools[J].Food&Chemical Toxicology An International Journal Published for the British Industrial Biological Research Association,2011,49(11):2954-2960.
[22]SORDET C,CULERRIER R,GRANIER C,et al.Expression of Jug r 1,the 2S albumin allergen from walnut(Juglans regia),as a correctly folded and functional recombinant protein[J].Peptides,2009,30(7):1213-1221.
[23]蔡秋凤,王锡昌,郭城,等.加工处理对鲢小清蛋白免疫活性的影响[J].水产学报,2013,37(2):297-302.
[24]GROZDANOVIC M,POPOVIC M,POLOVIC N,et al.Evaluation of Ig E reactivity of active and thermally inactivated actinidin,a biomarker of kiwifruit allergy[J].Food and Chemical Toxicology,2012,50(3/4):1013-1018.
[25]郑礼娜,林洪,刘一璇,等.不同热加工方式对刀额新对虾过敏原活性的影响[J].水产学报,2011,35(2):466-471.
[26]TONG P,GAO J Y,CHEN H B,et al.Effect of heat treatment on the potential allergenicity and conformational structure of egg allergen ovotransferrin[J].Food Chemistry,2012,131(2):603-610.
[27]CABANILLAS B,MALEKI S J,RODR魱GUEZ J,et al.Heat and pressure treatments effects on peanut allergenicity[J].Food Chemistry,2012,132(1):360-366.
[28]KORBER B,LABUTE M,YUSIM K.Immunoinformatics comes of age[J].Plos Computational Biology,2006,2(6):e71.
[29]SAHA S,RAGHAVA G P.Prediction of continuous B-cell epitopes in an antigen using recurrent neural network[J].Proteins,2006,65(1):4048.
[30]夏宏林,何颖,邹泽红,等.苹果过敏原Mal d4蛋白抗原表位预测及交叉反应分析[J].中国免疫学杂志,2012(1):57-61.
[31]李江英,白雪娟,梁艳,等.用DNAStar软件预测Rv1410c结核分枝杆菌蛋白抗原表位[J].细胞与分子免疫学杂志,2015,31(4):474-477.
[32]李海侠,毛旭虎.蛋白质抗原表位研究进展[J].微生物学免疫学进展,2007,35(1):54-58.
[33]LEE J M,KIM E J,KWON D G,et al.Clinical characteristics of walnut allergy and evaluation of cross-reactivity between walnut and peanut in children under 4 years of age[J].Pediatric Allergy&Respiratory Disease,2011,21(4):261-268.