Skoczowski Andrzej, Obtułowicz Krystyna, Czarnobilska Ewa, Dyga Wojciech, Mazur Marcel, Stawoska Iwona, Waga Jacek
Institute of Biology, Pedagogical University of Cracow, Kraków, Poland.
Department of Clinical and Environmental Allergology, Collegium Medicum, Jagiellonian University, Kraków, Poland.
Ann Agric Environ Med. 2017 May 11;24(2):229-236. doi: 10.5604/12321966.1233572.
[b]Abstract Introduction and objective[/b]. Gluten proteins (gliadins and glutenins) are polymorphic wheat storage proteins of allergenic properties. Significant differences in chemical composition between both protein groups allow to expect highly specific immunological response of individual subunits and fractions in reactions with IgE sera of people allergic to wheat. The aim of these studies was to identify and characterize the most allergenic gluten proteins (GP) and nongluten proteins (NGP) occurred in two closely related wheat hybrid genotypes. [b]Materials and method.[/b] 3xC and 3xN wheat hybrids, which differ strongly in regard of gliadin composition, were analyzed. Seven people manifesting different symptoms of wheat allergy donated sera for the experiment. The technique of immunoblotting after SDS-PAGE was used for identification of allergenic subunits and fractions among GP and NGP. Immunologically active protein bands were visualized by chemiluminescence. [b]Results[/b]. Great variation of immunodetection spectra was observed. Results of immunoblotting showed LMW glutenins to be of highest, gliadins of medium, while NGP of lowest allergenicity for selected patients. The 43-kDa and 47-kDa LMW glutenin subunits, 40-kDa and 43-kDa γ-gliadin fractions and 49-kDa NGP can be considered as the most immunoreactive among all protein bands [b]separated by SDS-PAGE.
[/b] The observed differentiation of immunodetection spectra allows to model highly specific IgE-binding profiles of allergenic wheat proteins attributed to individual patients with symptoms of gluten intolerance. Highly immunoreactive subunits and fractions among GP and NGP were identified. The observed immunoreactivity of 49 kDa NGP is worth to emphasize, as it has never been reported as wheat allergenic protein before.
[b]摘要 引言与目的[/b]。麸质蛋白（醇溶蛋白和谷蛋白）是具有致敏特性的多态性小麦贮藏蛋白。这两类蛋白在化学组成上存在显著差异，这使得人们预期个体亚基和组分在与对小麦过敏人群的IgE血清反应时会产生高度特异性的免疫反应。这些研究的目的是鉴定和表征在两种密切相关的小麦杂交基因型中出现的最具致敏性的麸质蛋白（GP）和非麸质蛋白（NGP）。[b]材料与方法[/b]。分析了醇溶蛋白组成差异很大的3xC和3xN小麦杂交种。7名表现出不同小麦过敏症状的人捐献血清用于实验。采用SDS-PAGE后的免疫印迹技术鉴定GP和NGP中的致敏亚基和组分。通过化学发光使具有免疫活性的蛋白条带可视化。[b]结果[/b]。观察到免疫检测谱有很大差异。免疫印迹结果显示，对于选定的患者，低分子量谷蛋白的致敏性最高，醇溶蛋白的致敏性中等，而非麸质蛋白的致敏性最低。在SDS-PAGE分离的所有蛋白条带中，43-kDa和47-kDa低分子量谷蛋白亚基、40-kDa和43-kDa γ-醇溶蛋白组分以及49-kDa NGP可被认为是免疫反应性最强的。
[/b]观察到的免疫检测谱差异使得能够模拟归因于有麸质不耐受症状个体患者的致敏小麦蛋白的高度特异性IgE结合谱。鉴定出了GP和NGP中免疫反应性强的亚基和组分。49 kDa NGP观察到的免疫反应性值得强调，因为它以前从未被报道为小麦致敏蛋白。
