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番茄肌动蛋白结合蛋白Lyc e 1:IgE交叉反应性和致敏效力。

Tomato profilin Lyc e 1: IgE cross-reactivity and allergenic potency.

作者信息

Westphal S, Kempf W, Foetisch K, Retzek M, Vieths S, Scheurer S

机构信息

Department of Allergology, Paul-Ehrlich-Institut, Langen, Germany.

出版信息

Allergy. 2004 May;59(5):526-32. doi: 10.1046/j.1398-9995.2003.00413.x.

Abstract

BACKGROUND

To date, very little data are available about the nature of tomato allergens. Immunoglobulin E (IgE) cross-reactive profilins have been suggested to account for allergic symptoms in patients suffering from tomato allergy.

METHODS

The cDNA of tomato profilin was amplified by reversely transcribed polymerase chain reaction (RT-PCR) from total RNA extracted from ripe tomato fruit. The gene was cloned into the pET101D expression plasmid and the protein was produced in Escherichia coli BL21. Purification was performed via poly-l-proline (PLP) affinity chromatography. IgE reactivity of recombinant tomato profilin was investigated by immunoblot and enzyme-linked immunosorbent assay. IgE-inhibition studies were performed to analyse cross-reactivity with other profilins. To determine the allergenic activity of the recombinant protein, basophil histamine release assays using sera of patients with adverse reactions to tomato were performed.

RESULTS

Profilin was identified as a new minor allergen in tomato fruits. The recombinant tomato profilin comprises 131 amino acids and high sequence identity to other allergenic food and pollen profilins. It was shown to be IgE-reactive with a prevalence of 22% (11/50) in tomato-allergic patients. In patients with tomato allergy and multiple sensitization to other foods and birch pollen, IgE directed against tomato profilin showed a strong cross-reactivity with profilins from plant food sources and birch pollen. The tomato profilin was able to induce mediator release from human basophils.

CONCLUSION

The tomato profilin is a minor allergen in tomato fruit. Thus, it shows biological activity, as confirmed by in vitro histamine release assays with human basophils and thereby has the potential to account for clinical symptoms in tomato-allergic patients.

摘要

背景

迄今为止,关于番茄过敏原的性质,可获得的数据非常少。免疫球蛋白E(IgE)交叉反应性肌动蛋白单体结合蛋白被认为是导致番茄过敏患者出现过敏症状的原因。

方法

通过逆转录聚合酶链反应(RT-PCR)从成熟番茄果实中提取的总RNA中扩增番茄肌动蛋白单体结合蛋白的cDNA。将该基因克隆到pET101D表达质粒中,并在大肠杆菌BL21中表达该蛋白。通过聚-L-脯氨酸(PLP)亲和层析进行纯化。通过免疫印迹和酶联免疫吸附测定研究重组番茄肌动蛋白单体结合蛋白的IgE反应性。进行IgE抑制研究以分析与其他肌动蛋白单体结合蛋白的交叉反应性。为了确定重组蛋白的致敏活性,使用对番茄有不良反应患者的血清进行嗜碱性粒细胞组胺释放试验。

结果

肌动蛋白单体结合蛋白被鉴定为番茄果实中的一种新的次要过敏原。重组番茄肌动蛋白单体结合蛋白由131个氨基酸组成,与其他致敏性食物和花粉肌动蛋白单体结合蛋白具有高度的序列同一性。在番茄过敏患者中,其显示出IgE反应性,患病率为22%(11/50)。在对番茄过敏且对其他食物和桦树花粉多重致敏的患者中,针对番茄肌动蛋白单体结合蛋白的IgE与来自植物性食物来源和桦树花粉的肌动蛋白单体结合蛋白显示出强烈的交叉反应性。番茄肌动蛋白单体结合蛋白能够诱导人嗜碱性粒细胞释放介质。

结论

番茄肌动蛋白单体结合蛋白是番茄果实中的一种次要过敏原。因此,它具有生物学活性,人体嗜碱性粒细胞体外组胺释放试验证实了这一点,从而有可能导致番茄过敏患者出现临床症状。

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