A novel allergen Tab y 1 with inhibitory activity of platelet aggregation from salivary glands of horseflies
A novel allergen Tab y 1 with inhibitory activity of platelet aggregation from salivary glands of horseflies
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具有抑制马蝇唾液腺血小板聚集活性的新型过敏原 表 1
DOI：
10.1111/j.1398-9995.2011.02683.x
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发表时间：
2011-11-01
期刊：
影响因子：
12.4
通讯作者：
Lai, R.
中科院分区：
文献类型：
--
作者：
An, S.;Ma, D.;Lai, R.
背景：马蝇叮咬可引起人体过敏反应。然而，我们对马蝇过敏原的了解仍然很少。目的：鉴定新型马蝇过敏原并对其性质进行表征。方法：采用凝胶过滤和离子交换色谱法，从麦夸马蝇的唾液腺中纯化出一种天然变应原蛋白Tab y 1 （apyrase）。通过Edman降解和cDNA克隆确定其序列。采用特异性IgE免疫印迹、嗜碱性粒细胞活化试验、皮肤点刺试验（SPT）和竞争性酶联免疫吸附试验（ELISA）评估其致敏性。结果：表y1在SDS-PAGE上显示一个70 kDa的单扩散带。70%（7/10）的马蝇过敏患者在SPT中Tab y 1检测呈阳性；81%（30/37）患者的血清在western blots上对Tab y 1有反应。纯化的Tab y 1在竞争性ELISA上降低了约42%的血清对马蝇唾液腺提取物的IgE反应性。表1上调CD63和CCR3在被动致敏的嗜碱性细胞上的表达，上调幅度约为4.9倍。表y1还显示酶水解ATP和ADP的活性，以及有效的抗血小板聚集和抗血栓活性。结论：本研究发现了一种新的马蝇主要过敏原Tab y 1，该过敏原具有抗血小板聚集和抗血栓活性，可能与Tab y 1帮助马蝇吸食宿主血液同时引起宿主过敏有关。
Background: Horsefly sting causes allergic reactions in human body. However, our knowledge on horsefly allergens remains poor.Objectives: To identify the novel horsefly allergens and characterize their properties.Methods: A native allergen protein Tab y 1 (apyrase) was purified from the salivary glands of the horsefly Tabanus yao Macquart by gel filtration and ion exchange chromatography. Its sequence was determined by Edman degradation and cDNA cloning. Its allergenicity was assessed by immunoblotting for specific IgE, basophil activation test, skin prick test (SPT), and competitive enzyme-linked immunosorbent assay (ELISA).Results: Tab y 1 showed a single diffusion band of 70 kDa on SDS-PAGE. Seventy percent (7/10) of patients with horsefly allergy tested positive to Tab y 1 in SPT; sera from 81% (30/37) of patients reacted to Tab y 1 on western blots. Purified Tab y 1 reduced approximately 42% sera IgE reactivity to horsefly salivary gland extract on a competitive ELISA. Tab y 1 upregulated the expression of CD63 and CCR3 on passively sensitized basophils by up to approximately 4.9-fold. Tab y 1 also showed enzymatic activity to hydrolyze ATP and ADP, and potent antiplatelet aggregation and antithrombotic activities.Conclusion: The current work identified a novel major allergen of horsefly, Tab y 1, with antiplatelet aggregation and antithrombotic activities, which implicates Tab y 1 in helping horseflies suck host blood, meanwhile causing allergy in their human hosts.