使用酶联免疫斑点分析（ELISPOT分析）在单细胞水平上检测细胞因子释放。

Measurement of cytokine release at the single cell level using the ELISPOT assay.

作者信息

Cox Josephine H, Ferrari Guido, Janetzki Sylvia

机构信息

U.S. Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, 13 Taft Court, Rockville, MD 20850, USA.

出版信息

Methods. 2006 Apr;38(4):274-82. doi: 10.1016/j.ymeth.2005.11.006.

DOI:10.1016/j.ymeth.2005.11.006

PMID:16473524

Abstract

The enzyme-linked immunospot (ELISPOT) assay took its design concept from traditional ELISA techniques and evolved over the years from a method for detecting antibodies secreted from B cells to a method for detecting cytokines or other soluble mediators secreted from a variety of different cell types. The ELISPOT assay allows the quantitative measurement of frequency of cytokine secreting cells at the single cell level directly ex vivo without elaborate in vitro expansion or manipulation of cell populations. The function of cells can be inferred from the pattern of cytokines secreted by cells in response to diverse antigenic stimuli and thus the ELISPOT assay has become a powerful method for monitoring immune responses in health and disease. The ELISPOT assay like the ELISA assay is relatively easy to perform and it has the promise of robustness, reliability, and reproducibility of performance for use as a diagnostic tool. The history, applications, validation process, and future challenges of the ELISPOT assay are discussed in this chapter.

摘要

酶联免疫斑点（ELISPOT）测定法的设计理念源自传统酶联免疫吸附测定（ELISA）技术，多年来已从一种检测B细胞分泌抗体的方法演变为一种检测多种不同细胞类型分泌的细胞因子或其他可溶性介质的方法。ELISPOT测定法能够在单细胞水平直接对分泌细胞因子的细胞频率进行定量测量，无需对细胞群体进行复杂的体外扩增或处理。细胞的功能可根据细胞对各种抗原刺激分泌的细胞因子模式来推断，因此ELISPOT测定法已成为监测健康和疾病中免疫反应的有力方法。与ELISA测定法一样，ELISPOT测定法相对易于操作，有望作为诊断工具具备稳健性、可靠性和性能可重复性。本章将讨论ELISPOT测定法的历史、应用、验证过程及未来挑战。