Pastorello Elide A, Farioli Laura, Pravettoni Valerio, Scibilia Joseph, Conti Amedeo, Fortunato Donatella, Borgonovo Linda, Bonomi Simona, Primavesi Laura, Ballmer-Weber Barbara
Unit of Allergology and Immunology-Niguarda Ca' Granda Hospital, Piazza Ospedale Maggiore 3, 20162 Milan, Italy.
Anal Bioanal Chem. 2009 Sep;395(1):93-102. doi: 10.1007/s00216-009-2945-z. Epub 2009 Aug 11.
Italian patients with maize anaphylaxis have been shown to have IgE toward two major maize allergens: an alpha-amylase inhibitor and a 9-kDa LTP. A complete study on maize food allergens in patients with positive maize double-blind, placebo-controlled food challenge (DBPCFC) is lacking. The objective was to utilize the three maize protein fractions to identify and characterize the most relevant IgE-binding proteins recognized by the sera of Italian and Swiss patients with either a positive maize-DBPCFC or a history of maize-induced anaphylaxis. Osborne's protein fractions of maize were extracted to obtain water-soluble, total zein, and total protein fractions. Protein IgE-binding capacity was investigated by SDS-PAGE immunoblotting using the sera from DBPCFC-positive patients and from patients with maize-induced anaphylaxis. Purified maize LTP was used to inhibit the IgE immunoblotting of the three protein fractions. IgE immunoblotting demonstrated that the 9-kDa LTP was recognized by all the Italian patients and by none of the Swiss patients. Other allergens were: 14-kDa alpha-amylase inhibitor, 30-kDa endochitinases A and -B, 19 kDa zein-beta precursor, and 26 kDa zein-alpha precursor; a newly described allergen, the globulin-2 precursor, identified in the total protein fraction. It is noteworthy that maize LTP and endochitinase were cross-reactive with grape LTP and one grape endochitinase. LTP was found to be the only major allergen in Italian patients with either positive maize challenge or a history of maize-induced anaphylaxis. We have identified other maize allergens in subjects with maize food allergy, as grape cross-reactive endochitinase, however, the clinical significance of these proteins needs to be investigated in larger groups of patients with allergy to these food items.
已证实,患有玉米过敏症的意大利患者体内存在针对两种主要玉米过敏原的IgE:一种α-淀粉酶抑制剂和一种9 kDa的脂质转移蛋白(LTP)。目前缺乏对玉米双盲、安慰剂对照食物激发试验(DBPCFC)呈阳性的患者体内玉米食物过敏原的全面研究。目的是利用三种玉米蛋白组分,鉴定和表征意大利和瑞士玉米DBPCFC呈阳性或有玉米诱发过敏反应病史的患者血清中识别的最相关IgE结合蛋白。提取玉米的奥斯本蛋白组分,以获得水溶性、总醇溶蛋白和总蛋白组分。使用DBPCFC呈阳性患者和有玉米诱发过敏反应患者的血清,通过SDS-PAGE免疫印迹法研究蛋白的IgE结合能力。使用纯化的玉米LTP抑制三种蛋白组分的IgE免疫印迹。IgE免疫印迹表明,所有意大利患者都能识别9 kDa的LTP,而所有瑞士患者均不能识别。其他过敏原包括:14 kDa的α-淀粉酶抑制剂、30 kDa的内切几丁质酶A和B、19 kDa的醇溶蛋白β前体和26 kDa的醇溶蛋白α前体;在总蛋白组分中鉴定出一种新描述的过敏原——球蛋白-2前体。值得注意的是,玉米LTP和内切几丁质酶与葡萄LTP和一种葡萄内切几丁质酶存在交叉反应。在玉米激发试验呈阳性或有玉米诱发过敏反应病史的意大利患者中,LTP被发现是唯一的主要过敏原。我们在患有玉米食物过敏的受试者中鉴定出了其他玉米过敏原,如葡萄交叉反应性内切几丁质酶,然而,这些蛋白的临床意义需要在更多对这些食物过敏的患者群体中进行研究。