Möbs Christian, Schmidt Thomas
Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany.
Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany.
J Invest Dermatol. 2016 Jun;136(6):e55-e59. doi: 10.1016/j.jid.2016.04.009.
The enzyme-linked immunospot (ELISPOT) assay allows characterization of single-cell immune responses through detection of secreted analytes. Although ELISPOT analysis shares similarities with ELISA, it has some essential differences. In general, the ELISPOT assay uses antibodies to capture and detect analytes of interest released by activated immune cells. Released analytes form specific antibody-antigen complexes and are visualized as spots of enzyme-substrate precipitates. These spots indicate both how many cells secrete the respective analyte and how much analyte is produced per individual cell. Initially developed for the detection of antibody-secreting cells, ELISPOT assays are now frequently performed both in the context of clinical diagnostics and in research on T-cell responses, in particular antigen-specific T-cell subpopulations, as related to allergy, cancer, infections, or autoimmune diseases. The one spot-one cell principle allows sensitive detection of specific and rare immune cell subsets. Here we present general principles, applications, and recent modifications of the ELISPOT technique.
酶联免疫斑点（ELISPOT）测定法可通过检测分泌的分析物来表征单细胞免疫反应。尽管ELISPOT分析与ELISA有相似之处，但也存在一些本质区别。一般来说，ELISPOT测定法使用抗体来捕获和检测活化免疫细胞释放的目标分析物。释放的分析物形成特定的抗体 - 抗原复合物，并可视化为酶 - 底物沉淀物的斑点。这些斑点既表明有多少细胞分泌相应的分析物，也表明每个细胞产生了多少分析物。ELISPOT测定法最初是为检测抗体分泌细胞而开发的，现在常用于临床诊断以及T细胞反应的研究，特别是与过敏、癌症、感染或自身免疫性疾病相关的抗原特异性T细胞亚群。一个斑点代表一个细胞的原则允许灵敏地检测特定且罕见的免疫细胞亚群。在此，我们介绍ELISPOT技术的一般原理、应用及最近的改进。
