Characterization of allergens secreted by Anisakis simplex parasite:: clinical relevance in comparison with somatic allergens
Characterization of allergens secreted by Anisakis simplex parasite:: clinical relevance in comparison with somatic allergens
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DOI：
10.1111/j.1365-2222.2004.01883.x
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发表时间：
2004-02-01
影响因子：
6.1
通讯作者：
Zubeldia, JM
中科院分区：
文献类型：
--
作者：
Baeza, ML;Rodríguez, A;Zubeldia, JM
研究背景单纯异尖线虫过敏反应的诊断方法基于幼虫的全身提取物的研究显然是不够的。目的研究寄生虫分泌的蛋白质的变应原性。与体细胞抗原的比较和确定其在过敏patients.Methods的临床重要性排泄/分泌（E/S）提取物的产生通过培养第三阶段的A.S.幼虫它被用来进行即时皮肤试验，并确定10例诊断为过敏的患者特异性IgE。将这两种测试与全身提取物（体细胞（S））获得的结果进行比较。通过免疫印迹法测定其过敏原的分子量（MW），并进行E/S蛋白的单盲安慰剂对照口服激发。最后探讨了过敏原对胃蛋白酶和酸性pH的抵抗力。幼虫分泌的过敏原比S提取物中存在的过敏原更有效。E/S分子在皮肤点刺试验中所产生的风团面积和用这些变应原测定特异性IgE（ELISA）所获得的吸光度比用S提取物所获得的吸光度大5.8倍。E/S提取物中72和56 kDa的MW变应原和S提取物中56、48和43 kDa的MW变应原被超过50%的患者识别。通过免疫印迹抑制研究揭示了它们之间的部分交叉反应性。所有患者经口激发E/S浸提液（高达479 μ g）均为阴性。用胃蛋白酶处理E/S蛋白可抑制E/S过敏原与特异性IgE的结合。酸性pH不影响IgE与E/S提取物的总体结合。在pH 4和2时，A.s.分别降低15.23%和19.96%，但差异无统计学意义。分泌比体细胞抗原更有效的过敏原，应用于诊断程序。这些过敏原被胃蛋白酶灭活，这支持了活幼虫是诱发大多数患者过敏反应所必需的理论。
Background Diagnostic methods for the study of allergic reactions to Anisakis simplex (A.s.) based on whole-body extracts of the larva are clearly insufficient.Objectives To study the allergenicity of the proteins secreted by the parasite. Comparison with somatic antigens and determination of their clinical importance in allergic patients were also addressed.Methods An excretory/secretory (E/S) extract was produced by culturing third-stage A.s. larvae. It was used to perform immediate skin tests and to determine specific IgE in 10 patients diagnosed with allergy to A.s. Both tests were compared with the results obtained with the whole-body extract (somatic (S)). The molecular weight (MW) of their allergens was determined by immunoblotting, and a single-blind placebo-controlled oral challenge with E/S proteins was performed. Finally, allergens' resistance to gastric pepsin and acid pH was explored.Results A.s. larvae secreted allergens more potent than those present in the S extract. The skin prick test wheal area produced by E/S molecules and the absorbance obtained in the determination of specific IgE with these allergens (ELISA) were 5.8 times bigger than those obtained with S extract. MW allergens of 72 and 56 kDa in E/S extracts and those of 56, 48 and 43 kDa in S extract were recognized by more than 50% of the patients. Partial cross-reactivity between them was revealed by immunoblotting inhibition studies. Oral challenge with E/S extract (up to 479 mug) was negative in all the patients. Treatment of E/S proteins with gastric pepsin inhibited the binding of the E/S allergens for specific IgE. The acid pH did not affect the overall binding of IgE to E/S extract. It decreased by 15.23% and 19.96% at pH 4 and 2, but the difference was not statistically significant.Conclusion A.s. secretes allergens more potent than somatic antigens and should be used in the diagnostic procedures. These allergens are inactivated by the pepsin, which supports the theory that live larva is necessary to induce an allergic reaction in most of the patients.