Localization of Four Allergens in Artemisia Pollen by Immunofluorescent Antibodies
Localization of Four Allergens in Artemisia Pollen by Immunofluorescent Antibodies
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DOI：
10.1159/000497321
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发表时间：
2019-01-01
影响因子：
2.8
通讯作者：
van Ree, Ronald
中科院分区：
文献类型：
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作者：
Gao, Zhong-Shan;Fu, Wan-Yi;van Ree, Ronald
背景：青蒿花粉极有可能诱发过敏症状。已经鉴定出七种过敏原成分，但只有Art v 7被定位在花粉粒中。方法：采用艾叶属（a.a argyi）和黄花艾属（a.a annua）两种中国艾属植物的花粉提取物免疫BALB/c小鼠。采用重组Art v 1和Art v 3过敏原筛选特异性单克隆抗体（mab）。用3个单抗纯化天然过敏原，然后用质谱法分析。如前所述，用合成的Art和7的3个肽段免疫家兔获得了多克隆抗体。采用常规组织学方法对4种艾属（a.a argyi, a.a annua, a.capilaris和a.sieversiana）的花粉进行观察，并通过荧光和共聚焦激光显微镜记录过敏原图像。结果：获得2个针对Art v 1、1个针对Art v 2、4个针对Art v 3同源物的特异性单克隆抗体。Art v 1和Art v 3同源蛋白主要位于花粉壁上，Art v 7同源蛋白定位于细胞核周围的细胞内。Art v2同源蛋白的位置在不同物种间存在差异，在黄花a.a annua和银叶a.a argyi中位于细胞核周围的细胞内，而在毛毛a.a和sieversiana中位于花粉壁和细胞核周围。结论：4种艾草变应原均定位于花粉中，Art v1和Art v3变应原主要位于花粉壁。
Background:Artemisia pollens have a high potential to induce allergic symptoms. Seven allergen components have been identified, but only Art v 7 has been localized in the pollen grain. This study aimed to localize the allergens in the pollen grains of 4 Artemisia spp. Methods: Pollen extracts from 2 Chinese Artemisia spp., A. argyi and A. annua, were used to immunize BALB/c mice. Recombinant Art v 1 and Art v 3 allergens were used to select specific monoclonal antibodies (mAbs). Three mAbs were used to purify the natural allergens and were then analyzed by mass spectrometry. As reported previously, polyclonal antibodies were obtained from rabbits immunized with 3 synthesized peptides of Art an 7. Using conventional histology procedures with pollens from 4 Artemisia spp. (A. argyi, A.annua, A. capilaris, and A. sieversiana), allergen images were observed and recorded by fluorescence and confocal laser microscopy. Results: We obtained 2 specific mAbs against Art v 1, 1 against Art v 2, and 4 against Art v 3 homologs. The Art v 1 and Art v 3 homologs were mainly located on the pollen walls, and the Art v 7 homologous protein was localized intracellularly around nuclei. The location of the Art v 2 homologous protein varied across species, being intracellular around nuclei for A. annua and A.argyi, and in both the pollen wall and around nuclei for A. capilaris and A.sieversiana. Conclusions: Four mugwort allergens were localized in the pollen, and the major Art v 1 and Art v 3 allergens were located mainly in the pollen wall.