致敏小鼠血清代替过敏患者血清筛选Jug r 1 IgE线性抗原表位的可行性研究
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摘要
目的:利用核桃过敏患者和核桃过敏原Jug r 1致敏小鼠血清筛选Jug r 1的IgE线性抗原表位,比较两者所筛选的表位的差异性,为今后利用小鼠血清替代人血清筛选食物过敏原表位提供理论依据,对食物过敏原研究及治疗具有重要意义。方法:利用固相合成肽法合成44个覆盖了Jug r 1一级序列的重叠短肽,通过圆点印迹试验利用核桃过敏患者血清和核桃过敏原Jug r 1致敏小鼠血清分别筛选出IgE抗原表位。结果:核桃过敏患者可以识别的IgE结合的线性抗原表位分别是肽段~1AALLVALLFVANAAA~(15)、4LVALLFVANAAAFRT18、~(16)FRTTITTMEIDEDID~(30)及~(125)CGISSQRCEIRRSWF~(139);核桃过敏原Jug r 1致敏小鼠所识别的抗原表位分别是1AALLVALLFVANAAA15和~(125)CGISSQRCEIRRSWF~(139)。结论:通过小鼠过敏血清筛选出的两个IgE线性抗原表位存在于过敏患者血清筛选出的4个IgE抗原表位中,提示在利用致敏小鼠血清代替过敏患者血清筛选IgE抗原表位时,可能出现假阴性结果,即有部分IgE线性抗原表位没有被识别。
Objective: The aim of this study was to screen the IgE linear Epitope of walnut major food allergen Jug r 1, using sensitized mice sera and pools of sera from walnut-allergic patients, and to compare the differences of the selected epitopes. Methods: We synthesized 44 solid-phase synthetic overlapping peptides which can cover the primary sequence of Jug r 1 to map the linear epitope of Jug r 1 with the Pools of sera from walnut-allergic patients and sensitized mice. Results: The results showed that peptide ~1AALLVALLFVANAAA~(15)、~4LVALLFVANAAA~(18),~(16)FRTTITTMEIDEDID~(30) and ~(125)CGISSQRCEIRRSWF~(139) can bind the specific IgE antibody from the patients. Peptides ~1AALLVALLFVANAAA~(15) and ~(125)CGISSQRCEIRRSWF~(139) can bind the specific IgE antibody from the sensitized mice. Conclusions: The IgE linear epitopes screened by sensitized mice were found in the four IgE epitopes screened by sera from walnut-allergic patients, suggesting that when IgE epitopes were screened by sera from sensitized mouse instead of from allergic patients, the tese may give us a false and negative result, that there will be part of the IgE linear epitopes are not recognized.
Objective: The aim of this study was to screen the IgE linear Epitope of walnut major food allergen Jug r 1, using sensitized mice sera and pools of sera from walnut-allergic patients, and to compare the differences of the selected epitopes. Methods: We synthesized 44 solid-phase synthetic overlapping peptides which can cover the primary sequence of Jug r 1 to map the linear epitope of Jug r 1 with the Pools of sera from walnut-allergic patients and sensitized mice. Results: The results showed that peptide ~1AALLVALLFVANAAA~(15)、~4LVALLFVANAAA~(18),~(16)FRTTITTMEIDEDID~(30) and ~(125)CGISSQRCEIRRSWF~(139) can bind the specific IgE antibody from the patients. Peptides ~1AALLVALLFVANAAA~(15) and ~(125)CGISSQRCEIRRSWF~(139) can bind the specific IgE antibody from the sensitized mice. Conclusions: The IgE linear epitopes screened by sensitized mice were found in the four IgE epitopes screened by sera from walnut-allergic patients, suggesting that when IgE epitopes were screened by sera from sensitized mouse instead of from allergic patients, the tese may give us a false and negative result, that there will be part of the IgE linear epitopes are not recognized.
引文
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[13]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.
[14]SAHA S,RAGHAVA G P.Prediction of continuous B-cell epitopes in an antigen using recurrent neural network[J].Proteins Structure Function&Bioinformatics,2006,65(1):40-48.
[15]ROBOTHAM J M,TEUBER S S,SATHE S K,et al.Linear IgE epitope mapping of the English walnut(Juglans regia)major food allergen,Jug r 1[J].Journal of Allergy&Clinical Immunology,2002,109(1):143-149.
[16]卢桂兰.北京地区人偏肺病毒全基因组序列、血清流行率分析及B细胞表位研究[D].北京:北京协和医学院,2009.
[17]DENERYPAPINI S,BODINIER M,PINEAU F,et al.Immunoglobulin-E-binding epitopes of wheat allergens in patients with food allergy to wheat and in mice experimentally sensitized to wheat proteins[J].Clinical&Experimental Allergy,2011,41(10):1478-1492.
[18]BATTAIS F,PINEAU F,POPINEAU Y,et al.Food allergy to wheat:identification of immunogloglin E and immunoglobulin G-binding proteins with sequential extracts and purified proteins from wheat flour[J].Clinical&Experimental Allergy,2003,33(7):962-970.
[2]宋帅,李春玲,贾爱卿,等.抗原表位研究方法进展[J].动物医学进展,2010,31(12):87-91.
[3]文雪霞,陈化兰,熊永忠,等.抗原表位鉴定方法的研究进展[J].中国畜牧兽医,2012,39(7):66-70.
[4]王艳华,张德林,殷宏,等.抗原表位预测方法的研究进展[J].中国兽医科学,2009,39(10):938-940.
[5]吴敬,吴梧桐.B细胞蛋白质抗原表位的研究方法进展[J].药物生物技术,2000,7(4):239-242.
[6]JUNG G,BECK-SICKINGER A G.Multiple peptide synthesis methods and their applications[J].Angewandte Chemie International Edition,1992,23(4):317-317.
[7]PAVI M R,SAWYER T K,MOOS W H.The generation of molecular diversity[J].Bioorganic&Medicinal Chemistry Letters,1993,3(3):387-396.
[8]FAO J,CONSULTATION W H O E,DERIVED F.Evaluation of allergenicity of genetically modified foods[J].Pediatric Allergy and Respiratory Disease,2008,18(4):292-304.
[9]刘光明,沈苑,曹敏杰,梁银龙,杨景成,苏文金.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.
[10]BRUIJNZEEL-KOOMEN C,ORTOLANI C,AAS K,et al.Adverse reactions to food[J].Allergy,1995,90(8):623-635.
[11]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.
[12]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.
[13]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.
[14]SAHA S,RAGHAVA G P.Prediction of continuous B-cell epitopes in an antigen using recurrent neural network[J].Proteins Structure Function&Bioinformatics,2006,65(1):40-48.
[15]ROBOTHAM J M,TEUBER S S,SATHE S K,et al.Linear IgE epitope mapping of the English walnut(Juglans regia)major food allergen,Jug r 1[J].Journal of Allergy&Clinical Immunology,2002,109(1):143-149.
[16]卢桂兰.北京地区人偏肺病毒全基因组序列、血清流行率分析及B细胞表位研究[D].北京:北京协和医学院,2009.
[17]DENERYPAPINI S,BODINIER M,PINEAU F,et al.Immunoglobulin-E-binding epitopes of wheat allergens in patients with food allergy to wheat and in mice experimentally sensitized to wheat proteins[J].Clinical&Experimental Allergy,2011,41(10):1478-1492.
[18]BATTAIS F,PINEAU F,POPINEAU Y,et al.Food allergy to wheat:identification of immunogloglin E and immunoglobulin G-binding proteins with sequential extracts and purified proteins from wheat flour[J].Clinical&Experimental Allergy,2003,33(7):962-970.
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