Ishii Soichiro, Koga Yuki, Yokooji Tomoharu, Kakino Misaki, Ogino Ryohei, Taogoshi Takanori, Matsuo Hiroaki
Department of Pharmaceutical Services, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8551, Japan.
Department of Frontier Science for Pharmacotherapy, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8553, Japan.
Foods. 2024 Aug 27;13(17):2713. doi: 10.3390/foods13172713.
The specific detection of serum IgE antibodies specific to allergens (sIgE Abs) that can crosslink the plural high-affinity IgE receptor (FcεRIα) molecules on the surface of mast cells or basophils with a multivalent allergen can reduce the false-positive diagnoses observed in chemiluminescent and fluorescence enzyme immunoassays for type-I allergic patients. In this study, we detected sIgE Abs to the egg-allergen ovalbumin (OVA) and the wheat-allergen gluten in the sera of rats sensitized with each allergen using an amplified luminescence proximity homogeneous assay by crosslinking (AlphaCL). OVA and gluten were reacted with each sIgE Ab in the sera. Then, acceptor and donor beads labeled with the human FcεRIα were added to the reacted solution. The luminescence intensity for anti-OVA IgE Abs in the sera with the removal of IgG Abs was observed in five of seven (71.4%) of the sensitized rats, whereas no signals were observed in any of the unsensitized rats. The AlphaCL could also detect anti-gluten sIgE Abs in the sera of sensitized rats, but not of unsensitized rats. In conclusion, we successfully detected sIgE Abs in the sera of rats sensitized to two allergens using the AlphaCL. This detection method has the potential to be used as a new diagnostic tool for type-I allergic patients.
能够通过多价变应原使肥大细胞或嗜碱性粒细胞表面的多个高亲和力IgE受体（FcεRIα）分子发生交联的变应原特异性血清IgE抗体（sIgE Abs）的特异性检测，可减少I型过敏患者在化学发光和荧光酶免疫分析中出现的假阳性诊断。在本研究中，我们使用交联增强发光邻近均相分析（AlphaCL），检测了用每种变应原致敏的大鼠血清中针对卵清蛋白（OVA）这一鸡蛋变应原和麸质这一小麦变应原的sIgE Abs。OVA和麸质与血清中的每种sIgE Ab发生反应。然后，将标记有人FcεRIα的受体珠和供体珠加入到反应溶液中。在去除IgG抗体的情况下，七只致敏大鼠中有五只（71.4%）的血清中观察到了抗OVA IgE Abs的发光强度，而未致敏大鼠中均未观察到信号。AlphaCL还能检测致敏大鼠血清中的抗麸质sIgE Abs，但不能检测未致敏大鼠血清中的抗麸质sIgE Abs。总之，我们使用AlphaCL成功检测了对两种变应原致敏的大鼠血清中的sIgE Abs。这种检测方法有可能用作I型过敏患者的一种新型诊断工具。
