Department of Pharmaceutical Services, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
Department of Pharmaceutical Services, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan; Department of Frontier Science for Pharmacotherapy, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
Allergol Int. 2022 Jan;71(1):94-102. doi: 10.1016/j.alit.2021.08.007. Epub 2021 Aug 28.
Autoantibodies (AAbs) against immunoglobulin E (IgE) antibodies (Abs) and their high-affinity receptor alpha subunits (FcεRIα) are key factors in the elicitation of type IIb autoimmune chronic spontaneous urticaria (type IIb aiCSU). In this study, we aimed to develop a new method to detect functional anti-FcεRIα and anti-IgE AAbs, which can crosslink the plural FcεRІα molecules and IgE Abs on the surface of mast cells and basophils, in sera from aiCSU patients using the amplified luminescence proximity homogeneous assay (Alpha).
Sera were obtained from 14 aiCSU patients, as diagnosed by recurrent chronic spontaneous urticaria episodes and positive results for the autologous serum skin test and/or histamine release test (HRT). The AAbs to FcεRIα and IgE Abs were determined in sera from aiCSU patients using enzyme-linked immunosorbent assay (ELISA) and Alpha by cross-linking (AlphaCL) of IgE Abs and/or FcεRІα.
Serum anti-FcεRIα and anti-IgE AAb levels were not significantly different between aiCSU patients and healthy subjects in ELISA. Anti-FcεRIα AAbs were detected in 10 of 14 aiCSU patients who displayed positive (5/5) and negative (5/9) results in the HRT for anti-FcεRIα AAbs by AlphaCL, whereas no signals were observed in healthy subjects. Additionally, anti-IgE AAbs were detected in two of four aiCSU patients who displayed positive results in the HRT for anti-IgE AAbs.
A new assay method using AlphaCL can detect anti-FcεRIα and anti-IgE AAbs with FcεRIα- and IgE-crosslinking abilities in sera from aiCSU patients. This simple and practical assay method may be available as a diagnostic tool for urticaria patients.
针对免疫球蛋白 E（IgE）抗体和其高亲和力受体 alpha 亚基（FcεRIα）的自身抗体（AAbs）是引发 IIb 型自身免疫性慢性自发性荨麻疹（IIb 型 aiCSU）的关键因素。在这项研究中，我们旨在开发一种新方法，使用放大发光接近均相测定法（Alpha），检测来自 aiCSU 患者血清中能够交联肥大细胞和嗜碱性粒细胞表面的多个 FcεRІα 分子和 IgE Ab 的功能性抗-FcεRIα 和抗-IgE AAbs。
从 14 名 aiCSU 患者中获得血清，这些患者的诊断依据是反复发作的慢性自发性荨麻疹发作以及自体血清皮肤试验和/或组胺释放试验（HRT）阳性。使用酶联免疫吸附试验（ELISA）和 Alpha 通过交联 IgE Abs 和/或 FcεRІα 来测定 aiCSU 患者血清中的抗-FcεRIα 和抗-IgE Ab。
在 ELISA 中，aiCSU 患者和健康受试者的血清抗-FcεRIα 和抗-IgE Ab 水平没有显著差异。在 AlphaCL 中，抗-FcεRIα Ab 检测到 14 名 aiCSU 患者中的 10 名（5/5）和 5 名（9/9）HRT 阳性的 aiCSU 患者（5/5）和 9 名（9/9）HRT 阴性的 aiCSU 患者（5/9）存在抗-FcεRIα Ab，而在健康受试者中则没有观察到信号。此外，在 HRT 中抗-IgE Ab 检测到 4 名 aiCSU 患者中的 2 名（2/4）为阳性。
使用 AlphaCL 的新测定方法可以检测来自 aiCSU 患者血清中具有 FcεRIα 和 IgE 交联能力的抗-FcεRIα 和抗-IgE AAbs。这种简单实用的测定方法可能成为荨麻疹患者的一种诊断工具。
