IDENTIFICATION OF A MAJOR PEANUT ALLERGEN, ARA-H-I, IN PATIENTS WITH ATOPIC-DERMATITIS AND POSITIVE PEANUT CHALLENGES
IDENTIFICATION OF A MAJOR PEANUT ALLERGEN, ARA-H-I, IN PATIENTS WITH ATOPIC-DERMATITIS AND POSITIVE PEANUT CHALLENGES
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DOI：
10.1016/0091-6749(91)90325-i
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发表时间：
1991-08-01
影响因子：
14.2
通讯作者：
OBRIEN, T
中科院分区：
文献类型：
--
作者：
BURKS, AW;WILLIAMS, LW;OBRIEN, T
花生是食物引起的速发型超敏反应最常见的原因之一。使用9名特应性皮炎患者的血清（这些患者在双盲、安慰剂对照的花生食物激发试验中呈阳性）来开始鉴定和纯化主要花生过敏原的过程。通过阴离子交换柱色谱法、十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳、酶联免疫吸附测定（ELISA）、薄层等电聚焦以及IgE特异性免疫印迹法完成了对一种主要花生过敏原的鉴定。阴离子交换色谱法显示出几个能与激发试验阳性患者血清中的IgE结合的组分。通过在ELISA和IgE特异性免疫印迹法中测量抗花生特异性IgE，我们通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳鉴定出一种具有两条考马斯亮蓝染色带的致敏成分，其平均分子量为63.5 kd。通过用个体血清进行IgE抗花生ELISA检测以及用混合血清进行ELISA抑制试验对该组分进行检测，我们确定该组分为一种主要过敏原。对这个63.5 kd组分进行薄层等电聚焦和免疫印迹分析，发现其等电点为4.55。根据国际免疫学会联合会过敏原命名小组委员会的过敏原命名法，这种过敏原被命名为Ara h I（花生）。
Peanuts are among the most common causes of immediate hypersensitivity reactions to foods. Serum from nine patients with atopic dermatitis and a positive double-blind, placebo-controlled, food challenge to peanut were used to begin the process of identification and purification of the major peanut allergens. Identification of a major peanut allergen was accomplished by use of anion-exchange column chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, ELISA, thin-layer isoelectric focusing, and IgE-specific immunoblotting. Anion-exchange chromatography revealed several fractions that bound IgE from the serum of the challenge-positive patient pool. By measuring antipeanut-specific IgE in the ELISA and in IgE-specific immunoblotting, we identified an allergenic component with two Coomassie brilliant blue staining bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a mean molecular weight of 63.5 kd. Examination of this fraction by the IgE antipeanut ELISA with individual serum and by the ELISA-inhibition assay with pooled serum, we identified this fraction as a major allergen. Thin-layer isoelectric focusing and immunoblotting of this 63.5 kd fraction revealed it to have an isoelectric point of 4.55. Based on allergen nomenclature of the IUIS Subcommittee for Allergen Nomenclature, this allergen is designated, Ara h I (Arachis hypogaea).