Division of Palynology and Environmental Biology, Department of Botany, Bose Institute, Calcutta, India.
Ann Allergy Asthma Immunol. 2009 Dec;103(6):515-24. doi: 10.1016/S1081-1206(10)60269-4.
Delonix regia and Peltophorum pterocarpum pollen are important aeroallergens for type 1 hypersensitivity in the tropics. The IgE-binding proteins of D regia and their cross-allergenity with P pterocarpum pollen have not been evaluated.
To isolate and characterize the IgE-binding proteins of D regia pollen for the first time and to investigate the cross-allergenity with P pterocarpum pollen belonging to the same family (Leguminosae).
Allergenic activities were determined by in vivo and in vitro analyses. Pollen extract was fractionated by a combination of 2 columns (diethyl amino ethyl Sephadex and Sephacryl S-200). Protein components were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, periodic acid-Schiff staining, and immunoblotting. In vitro inhibition tests were performed to evaluate the cross-reactivity.
The skin prick test results of the patients with respiratory allergies in Calcutta, India, showed 31.1% positivity with D regia pollen. Nine IgE-reactive protein components were found in the crude extract. An optimum IgE-reactive fraction was resolved into 4 subfractions. Subfraction A, which showed maximum IgE reactivity, contained 2 (96- and 66-kDa) IgE-reactive protein components. The 66-kDa component was found to be glycoprotein. Remarkable cross-reactivity between D regia and P pterocarpum pollen was found on IgE enzyme-linked immunosorbent assay inhibition and dot blotting. Shared IgE-binding components (66, 56, 32, 28, 25, and 23 kDa) were observed between D regia and P pterocarpum pollen extracts, whereas the 96- and 43-kDa components were specific to D regia.
The purification of the IgE-binding proteins and the identification of the shared/cross-reactive proteins in these taxonomically related pollen members should be helpful for the diagnosis and therapy of patients susceptible to these pollens.
在热带地区,无忧树花粉和紫檀花粉是 1 型超敏反应的重要过敏原。无忧树花粉的 IgE 结合蛋白及其与同科(豆科)紫檀花粉的交叉过敏原性尚未得到评估。
首次分离和鉴定无忧树花粉的 IgE 结合蛋白,并研究其与同科(豆科)紫檀花粉的交叉过敏原性。
通过体内和体外分析测定变应原活性。花粉提取物通过 2 种柱(二乙基氨基乙基 Sephadex 和 Sephacryl S-200)的组合进行分级。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、过碘酸-希夫染色和免疫印迹法对蛋白质成分进行表征。进行体外抑制试验以评估交叉反应性。
印度加尔各答的呼吸道过敏患者的皮肤点刺试验结果显示,31.1%的患者对无忧树花粉呈阳性。在粗提物中发现了 9 种 IgE 反应性蛋白成分。最佳 IgE 反应性部分进一步分离为 4 个亚部分。显示最大 IgE 反应性的亚部分 A 含有 2 个(96-和 66-kDa)IgE 反应性蛋白成分。发现 66-kDa 成分是糖蛋白。在 IgE 酶联免疫吸附试验抑制和点印迹中发现了无忧树和紫檀花粉之间的显著交叉反应性。在无忧树和紫檀花粉提取物之间观察到共享 IgE 结合成分(66、56、32、28、25 和 23 kDa),而 96-和 43-kDa 成分是无忧树特有的。
这些分类相关花粉成员的 IgE 结合蛋白的纯化以及共享/交叉反应性蛋白的鉴定,应该有助于这些花粉易感患者的诊断和治疗。