Department of Dermatology, Institute of Biomedical & Health Sciences, Hiroshima University, Hiroshima, Japan.
Unit of Biotechnology, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan.
Allergol Int. 2020 Apr;69(2):253-260. doi: 10.1016/j.alit.2019.09.007. Epub 2019 Nov 7.
Fish roe allergy is a common health problem in countries where sea food is a major part of the diet, such as Japan. β'-component (β'-c) in fish roe has been identified as a major antigen for patients who show hypersensitivity to various fish roes. However, little is known about causative antigens for patients reactive to fish roe of specific species.
Serum and basophils were obtained from patients who had reactivity to roes of Gadus chalcogrammus (GC) and/or other fish species. GC roe specific antigens were analyzed by immunoblotting, histamine release assay (HRA) and mass spectrometry. Recombinant-fragments of vitellogenin (Vg) were obtained by the Escherichia coli expression system.
Serum IgE of a patient with specific reactions to GC roe bound to 15, 28, 40 and 70 kDa-proteins in GC roe extract. Mass spectrometry analysis revealed that proteins in these bands contained fragments corresponding to Vg. Immunoblotting of Vg immunoprecipitated by rabbit anti-Vg antiserum from the extract revealed 15, 28 and 54 kDa fragments bound by the patient's IgE. These bindings were inhibited by the pretreatment of recombinant phosvitin (rPv) and β'-c (rβ'-c). Fractions obtained by native gel electrophoresis containing 15, 28 and 54 kDa proteins, but not the other fractions, induced significant histamine release from the patient's basophils. Sera of the other patients with GC roe specific-IgE showed IgE binding to rPv and/or rβ'-c.
The 15, 28 and 54 kDa-fragments of Vg which include structures of Pv and β'-c, could be antigens for GC roe specific type-I-hypersensitivity.
鱼卵过敏是在以海鲜为主要饮食的国家（如日本）常见的健康问题。β'-成分（β'-c）已被确定为对各种鱼卵过敏的患者的主要抗原。然而，对于对特定鱼类鱼卵反应的患者，导致抗原的原因知之甚少。
从对 Gadus chalcogrammus（GC）和/或其他鱼类鱼卵有反应的患者中获得血清和嗜碱性粒细胞。通过免疫印迹、组胺释放测定（HRA）和质谱分析来分析 GC 鱼卵特异性抗原。通过大肠杆菌表达系统获得卵黄蛋白（Vg）的重组片段。
对 GC 鱼卵有特异性反应的患者的血清 IgE 与 GC 鱼卵提取物中的 15、28、40 和 70 kDa 蛋白结合。质谱分析表明，这些带中的蛋白含有对应于 Vg 的片段。用兔抗 Vg 抗血清从提取物中免疫沉淀 Vg 后进行免疫印迹，显示与患者 IgE 结合的 15、28 和 54 kDa 片段。这些结合物可通过重组磷蛋白（rPv）和β'-c（rβ'-c）预处理来抑制。含有 15、28 和 54 kDa 蛋白的天然凝胶电泳获得的分数，但不是其他分数，可从患者嗜碱性粒细胞中诱导显著的组胺释放。其他具有 GC 鱼卵特异性 IgE 的患者的血清显示 IgE 与 rPv 和/或 rβ'-c 结合。
Vg 的 15、28 和 54 kDa 片段包含 Pv 和β'-c 的结构，可能是 GC 鱼卵特异性 I 型超敏反应的抗原。
