Department of Pharmaceutical Services, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8553, Japan.
Department of Frontier Science for Pharmacotherapy, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
Sci Rep. 2021 Mar 8;11(1):5400. doi: 10.1038/s41598-021-84752-2.
Shrimp is a causative food that elicits food-dependent exercise-induced anaphylaxis (FDEIA). In this study, we sought to identify IgE-binding allergens in patients with shrimp-FDEIA. Sera were obtained from eight patients with shrimp-FDEIA and two healthy control subjects. Proteins were extracted from four shrimp species by homogenization in Tris buffer. Immunoblot analysis revealed that IgE from patient sera bound strongly to a 70-kDa and a 43-kDa protein in a preparation of Tris-soluble extracts from Litopenaeus vannamei. Mass spectrometry identified the 70-kDa and 43-kDa proteins as a P75 homologue and fructose 1,6-bisphosphate aldolase (FBPA), respectively. To confirm that the putative shrimp allergens were specifically recognized by serum IgE from shrimp-FDEIA patients, the two proteins were purified by ammonium sulfate precipitation followed by reversed-phase HPLC and/or anion-exchange hydrophobic interaction chromatography and then subjected to immunoblot analysis. Purified P75 homologue and FBPA were positively bound by serum IgE from one and three, respectively, of the eight patients with shrimp-FDEIA, but not by sera from control subjects. Thus, P75 homologue and FBPA are identified as IgE-binding allergens for shrimp-FDEIA. These findings could be useful for the development of diagnostic tools and desensitization therapy for shrimp-FDEIA patients.
虾是一种诱发食物,可引发与食物相关的运动诱发过敏反应(FDEIA)。在这项研究中,我们试图确定虾-FDEIA 患者中的 IgE 结合过敏原。从 8 例虾-FDEIA 患者和 2 例健康对照中获得血清。通过在 Tris 缓冲液中匀浆从四种虾中提取蛋白质。免疫印迹分析显示,来自患者血清的 IgE 与 Tris 可溶提取物制剂中的 70 kDa 和 43 kDa 蛋白强烈结合。质谱鉴定 70 kDa 和 43 kDa 蛋白分别为 P75 同源物和果糖 1,6-二磷酸醛缩酶(FBPA)。为了确认假定的虾过敏原被虾-FDEIA 患者的血清 IgE 特异性识别,将两种蛋白质通过硫酸铵沉淀、反相 HPLC 和/或阴离子交换疏水性相互作用色谱进行纯化,然后进行免疫印迹分析。来自 8 例虾-FDEIA 患者中的 1 例和 3 例的血清 IgE 分别与纯化的 P75 同源物和 FBPA 阳性结合,但与对照血清无反应。因此,P75 同源物和 FBPA 被鉴定为虾-FDEIA 的 IgE 结合过敏原。这些发现可能对虾-FDEIA 患者的诊断工具和脱敏治疗的开发有用。