Mapping of the IgE and IgG4 sequential epitopes of milk allergens with a peptide microarray-based immunoassay
Mapping of the IgE and IgG4 sequential epitopes of milk allergens with a peptide microarray-based immunoassay
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DOI：
10.1016/j.jaci.2008.06.040
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发表时间：
2008-09-01
影响因子：
14.2
通讯作者：
Sampson, Hugh A.
中科院分区：
文献类型：
--
作者：
Cerecedo, Inmaculada;Zamora, Javier;Sampson, Hugh A.
背景：肽芯片分析是一种新的方法，可以提供有用的信息，对特定的过敏性质。目的：我们试图通过基于肽芯片的免疫分析，确定IgE和IgG4抗体结合α (s1)-、α (s2)-、β -和kappa-酪蛋白和β -乳球蛋白序列表位的特异性和多样性。方法：对31例ige介导的牛奶过敏患儿血清进行微阵列免疫分析，其中16例口服奶攻毒结果为阳性（即反应组），15例口服奶攻毒结果为阴性（即耐受组）。由20个氨基酸（aa）重叠17个（3-偏移）组成的肽库，对应于α (s1)-, α (s2)-， β -和kappa-酪蛋白和β -乳球蛋白的初级序列，在环氧涂层载玻片上打印。如果一个区域在统计学上与反应性组相关，并且被至少75%的反应性患者识别，则该区域被定义为表位。结果：该方法共鉴定出10个抗原表位：α (s1), AAs 28 ~ 50, 75%反应性，26.7%耐受性；a、2、AAs I ~ 20,75%反应性和13.3%耐受性；AAs 13 ~ 32, 75%反应性，26.7%耐受性；AAs 67 ~ 86, 75%反应性和33.3%耐受性；AAs 181 ~ 207, 75%反应性和20%耐受性；β -酪蛋白，AAs 25 ~ 50, 75%反应性和33.3%耐受性，AAs 52 ~ 74, 81.3%反应性和26.7%耐受性；AAs 154 ~ 173，反应性75%，耐受性33.3%；-乳球蛋白，AAs 58 ~ 77，反应性81.3%，耐受性40%；kappa-酪蛋白，AAs 34 ~ 53，反应性87.5%，耐受性40%。结论：几个区域被定义为表位，在反应性和耐受性患者之间表现出不同的识别模式。需要进一步的研究来验证这种测定在临床实践中的效用。
Background: Peptide microarray analysis is a novel method that can provide useful information on the nature of specific allergies.Objective: We sought to determine the specificity and diversity of IgE and IgG4 antibodies binding to sequential epitopes of alpha(s1)-, (alpha(s2)-, beta-, and kappa-caseins and beta-lactoglobulin by using a peptide microarray-based immunoassay.Methods: A microarray immunoassay was performed with sera from 31 children with IgE-mediated milk allergy (16 with positive oral milk challenge results [ie, the reactive group] and 15 with negative oral milk challenge results [ie, the tolerant group]). A library of peptides, consisting of 20 amino acids (AAs) overlapping by 17 (3-offset), corresponding to the primary sequences of alpha(s1)-, alpha(s2)-, beta-, and kappa-caseins and beta-lactoglobulin was printed on epoxy-coated slides. A region was defined as an epitope if it was statistically associated with reactive groups and recognized by at least 75% of reactive patients.Results: By using this method, a total of 10 epitopes were identified: alpha(s1), AAs 28 to 50, 75% reactive and 26.7% tolerant; a,2, AAs I to 20, 75% reactive and 13.3% tolerant; AAs 13 to 32, 75% reactive and 26.7% tolerant; AAs 67 to 86, 75% reactive and 33.3% tolerant, and AAs 181 to 207, 75% reactive and 20% tolerant; beta-casein, AAs 25 to 50, 75% reactive and 33.3% tolerant, AAs 52 to 74, 81.3% reactive and 26.7% tolerant; and AAs 154 to 173, 75% reactive and 33.3% tolerant; beta-lactoglobulin, AAs 58 to 77, 81.3% reactive and 40% tolerant; and kappa-casein, AAs 34 to 53, 87.5% reactive and 40% tolerant.Conclusion: Several regions have been defined as epitopes, which showed differential recognition patterns between reactive and tolerant patients. Further studies are needed to validate the utility of this assay in clinical practice.