Tundo Silvio, Lupi Roberta, Lafond Mickael, Giardina Thierry, Larré Colette, Denery-Papini Sandra, Morisset Martine, Kalunke Raviraj, Sestili Francesco, Masci Stefania
Department of Agriculture and Forest Science (DAFNE), University of Tuscia, Via S. C. de Lellis snc, 01100 Viterbo, Italy.
Present address: Department of Land, Environment, Agriculture and Forestry (TESAF), University of Padova, Viale dell'Università 16, 35020 Legnaro (PD), Italy.
Plants (Basel). 2018 Nov 16;7(4):101. doi: 10.3390/plants7040101.
Although wheat is a staple food for most of the human population, some of its components trigger adverse reactions. Among wheat components, the alpha-amylase/trypsin inhibitors (ATI) are important triggers of several allergies and activators of innate immunity. ATI are a group of exogenous protease inhibitors and include several polypeptides. The three ATI polypeptides named CM3, CM16 and 0.28 are considered major allergens, and might also play a role in other common wheat-related pathologies, such as Non Celiac Wheat Sensitivity and even Celiac Disease. On this basis, we pointed to obtain high amounts of them in purity and to evaluate their allergenicity potential. We thus isolated the mRNA corresponding to the three ATI genes CM3, CM16 and 0.28 from 28 days post-anthesis wheat kernels and the corresponding cDNAs were used for heterologous expression in . The three purified proteins were tested in degranulation assay against human sera of patients with food allergy to wheat. A large range of degranulation values was observed for each protein according to the sera tested. All of the three purified proteins CM3, CM16 and 0.28 were active as allergens because they were able to induce basophils degranulation on wheat allergic patients' sera, with the highest values of β-hexosaminidase release observed for CM3 protein.
尽管小麦是大多数人类的主食，但其某些成分会引发不良反应。在小麦成分中，α-淀粉酶/胰蛋白酶抑制剂（ATI）是几种过敏反应的重要触发因素和先天免疫的激活剂。ATI是一组外源性蛋白酶抑制剂，包括几种多肽。名为CM3、CM16和0.28的三种ATI多肽被认为是主要过敏原，它们也可能在其他常见的小麦相关病症中起作用，如非乳糜泻性小麦敏感甚至乳糜泻。在此基础上，我们旨在大量获取纯度高的它们，并评估其潜在的致敏性。因此，我们从开花后28天的小麦籽粒中分离出与三个ATI基因CM3、CM16和0.28相对应的mRNA，并将相应的cDNA用于在……中进行异源表达。对三种纯化的蛋白质进行了针对对小麦食物过敏患者血清的脱颗粒试验。根据所测试的血清，每种蛋白质都观察到了大范围的脱颗粒值。三种纯化的蛋白质CM3、CM16和0.28均具有过敏原活性，因为它们能够诱导小麦过敏患者血清中的嗜碱性粒细胞脱颗粒，其中CM3蛋白观察到的β-己糖胺酶释放值最高。
