Baghersad Ali, Shams Mohammad Hossein, Shahsavar Farhad, Sohrabi Seyyed Mohsen, Varzi Ali Mohammad, Baharvand Peyman Amanolahi, Eskandari Nahid, Darestani Nadia Ghasemi, Sereshki Hale Abdoli, Doosti Fatemeh, Assarehzadegan Mohammad-Ali
Hepatitis Research Center and Department of Medical Immunology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
Hepatitis Research Center and Department of Medical Immunology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
Int Immunopharmacol. 2023 Nov;124(Pt B):110953. doi: 10.1016/j.intimp.2023.110953. Epub 2023 Sep 25.
Pollens, particularly tree and plant pollens, are one of the major causes of allergic respiratory diseases worldwide. Allergy to pollens of different species of Salix trees has been reported in various regions of the world. The most common type of Salix tree in Iran is white willow (Salix alba).
This study aimed to identify and determine the immunochemical characteristics of allergenic proteins in S. alba tree pollen extract using SDS-PAGE and IgE- immunoblotting methods. Moreover, the cross-reaction pattern of the specific IgE antibody of S. alba tree pollen proteins with pollen allergens of common allergenic trees, i.e., Populus nigra (P. nigra), Cupressus sempervirens (C. sempervirens), Pinus brutia (P. brutia) and Platanus orientalis (P. orientalis) in the region was investigated.
The reaction of allergenic proteins in S. alba pollen extract with specific IgE antibodies in patients' sera was investigated using SDS-PAGE and IgE-immunoblotting methods. The cross-reaction of specific IgE antibodies of the proteins present in S. alba pollen extract with pollen allergens of common allergenic trees in the region was investigated using ELISA and immunoblotting inhibition methods. In silico methods such as phylogenetic tree drawing and alignment of amino acid sequences were used to examine the evolutionary relationship and homology structure of common allergenic proteins (Panallergens) responsible for cross reactions.
More than 11 protein bands binding to specific IgE antibodies in patients' sera with a molecular weight between 13 and 95 kDa were identified in the S. alba tree pollen extract. ELISA and immunoblotting inhibition results showed that P. nigra extract could inhibit the binding of IgE antibodies to S. alba pollen extract proteins to a greater extent than C. sempervirens, P. brutia, and P. orientalis tree extracts. In silico methods investigated the results of ELISA and immunoblotting inhibition methods. Moreover, a high structural homology and evolutionary relationship were observed between S. alba and P. nigra tree pollen panallergens.
In this study, it was found that more than 80 % of the sensitive patients who were examined had specific IgE antibodies reacting with the approximately a 15 kDa-protein present in the S. alba pollen extract. Furthermore, the specific IgE-binding proteins found in the pollens of S. alba and P. nigra trees had relative structural homology, and it is likely that if recombinant forms are produced, they can be used for diagnostic and therapeutic purposes for both of the trees.
花粉，尤其是树木和植物花粉，是全球过敏性呼吸道疾病的主要病因之一。世界各地区均有对不同种类柳树花粉过敏的报道。伊朗最常见的柳树品种是白柳（Salix alba）。
本研究旨在采用十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）和免疫球蛋白E（IgE）免疫印迹法，鉴定并确定白柳树花粉提取物中变应原蛋白的免疫化学特征。此外，还研究了白柳树花粉蛋白特异性IgE抗体与该地区常见致敏树木即黑杨（Populus nigra）、地中海柏木（Cupressus sempervirens）、土耳其松（Pinus brutia）和悬铃木（Platanus orientalis）花粉变应原的交叉反应模式。
采用SDS - PAGE和IgE免疫印迹法研究白柳花粉提取物中变应原蛋白与患者血清中特异性IgE抗体的反应。采用酶联免疫吸附测定（ELISA）和免疫印迹抑制法研究白柳花粉提取物中蛋白质的特异性IgE抗体与该地区常见致敏树木花粉变应原的交叉反应。采用系统发育树绘制和氨基酸序列比对等计算机模拟方法，研究负责交叉反应的常见致敏蛋白（泛变应原）的进化关系和同源结构。
在白柳树花粉提取物中鉴定出11条以上与患者血清中特异性IgE抗体结合的蛋白条带，分子量在13至95 kDa之间。ELISA和免疫印迹抑制结果表明，黑杨提取物比地中海柏木、土耳其松和悬铃木提取物更能抑制IgE抗体与白柳花粉提取物蛋白的结合。计算机模拟方法验证了ELISA和免疫印迹抑制方法的结果。此外，白柳和黑杨花粉泛变应原之间存在高度的结构同源性和进化关系。
本研究发现，超过80%接受检查的敏感患者具有与白柳花粉提取物中约15 kDa蛋白发生反应的特异性IgE抗体。此外，白柳和黑杨花粉中发现的特异性IgE结合蛋白具有相对的结构同源性，若生产重组形式，它们可能可用于这两种树的诊断和治疗目的。
