Identification of a 62-kDa major allergen from Artemisia pollen as a putative galactose oxidase
Identification of a 62-kDa major allergen from Artemisia pollen as a putative galactose oxidase
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从青蒿花粉中鉴定一个62-kDa的半乳糖氧化酶主要过敏原
DOI：
10.1111/all.13375
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发表时间：
2018-05-01
期刊：
影响因子：
12.4
通讯作者：
van Ree, R.
中科院分区：
文献类型：
--
作者：
Fu, W.;Gao, Z.;van Ree, R.
背景大约 20 年前，据报道一种 60 至 70 kDa 的蛋白质是艾蒿花粉的主要过敏原。本研究旨在鉴定和表征其分子特性。方法使用来自 113 名中国和 20 名荷兰蒿过敏/致敏受试者（及其池）的血清来鉴定 60 至 70 kDa 过敏原。通过免疫印迹比较七种蒿属植物的花粉提取物。使用青蒿花粉的转录组学和蛋白质组学（质谱法）来鉴定假定的 60 至 70 kDa 蒿过敏原。通过ImmunoCAP 评估天然纯化和重组过敏原的IgE 反应性。对 14 名中国蒿过敏患者进行了皮内纯化天然过敏原测试。结果免疫印迹显示所有七种蒿属植物均在 12 和 25kDa 处有两个主要条带，在 70kDa 处有一条弱条带。使用转录组学和蛋白质组学相结合的方法，青蒿花粉中的高分子量过敏原被证明是一种 62 kDa 的推定半乳糖氧化酶，具有推定的 N-糖基化位点。超过 94% 的蒿花粉过敏患者对该过敏原有 IgE 反应。尽管仅在少数（16%）和低得多的 IgE 水平下确认了对非糖基化重组版本的识别，但这种差异不能简单地用对天然过敏原上碳水化合物部分的反应性来解释。九名致敏患者中有五人的天然过敏原皮内测试呈阳性。结论先前报道的蒿花粉的 60-至 70-kDa 过敏原很可能是一种 62-kDa 推定的半乳糖氧化酶，此处指定为 Art 和 7。
BackgroundAround 20years ago, a 60- to 70-kDa protein was reported as a major allergen of mugwort (Artemisia vulgaris) pollen. This study was to identify and characterize its molecular properties.MethodsSera from 113 Chinese and 20 Dutch Artemisia-allergic/sensitized subjects (and pools thereof) were used to identify the 60- to 70-kDa allergen. Pollen extracts of seven Artemisia species were compared by immunoblotting. Transcriptomics and proteomics (mass spectrometry) of A.annua pollen were used to identify the putative 60- to 70-kDa Artemisia allergen. Both the natural purified and recombinant allergens were evaluated for IgE reactivity by ImmunoCAP. Fourteen Chinese Artemisia-allergic patients were tested intradermally with purified natural allergen.ResultsImmunoblots revealed two major bands at 12 and 25kDa, and a weak band at 70kDa for all seven Artemisia species. Using a combined transcriptomic and proteomic approach, the high molecular mass allergen in A.annua pollen was shown to be a 62-kDa putative galactose oxidase, with a putative N-glycosylation site. More than 94% of Artemisia pollen-allergic patients had IgE response to this allergen. Although recognition of a nonglycosylated recombinant version was only confirmed in a minority (16%) and at much lower IgE levels, this discrepancy cannot be explained simply by reactivity to the carbohydrate moiety on the natural allergen. Intradermal testing with the natural allergen was positive in five of nine sensitized patients.ConclusionsThe previously reported 60- to 70-kDa allergen of Artemisia pollen is most likely a 62-kDa putative galactose oxidase here designated Art an 7.