Comparison of the ELISPOT and cytokine flow cytometry assays for the enumeration of antigen-specific T cells
Comparison of the ELISPOT and cytokine flow cytometry assays for the enumeration of antigen-specific T cells
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DOI：
10.1016/j.jim.2003.09.001
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发表时间：
2003-12-01
影响因子：
2.2
通讯作者：
Sinclair, E
中科院分区：
文献类型：
--
作者：
Karlsson, AC;Martin, JN;Sinclair, E
近年来，基于细胞因子检测的方法的发展极大地促进了抗原特异性T细胞应答的计数。特别是，酶联免疫斑点（ELISPOT）和细胞因子流式细胞术（CFC）测定已变得流行。由于这两种检测方法可能会继续广泛使用，因此评价其结果是否具有可比性非常重要。在目前的研究中，我们比较了ELISPOT和CFC检测中获得的结果，使用对应于CMV和HIV-1蛋白的肽池在慢性HIV-1感染者。对肽库的T细胞应答的分析表明，CMV pp 65和HIN-1 Gag CFC和ELISPOT衍生的结果在统计学上相关。然而，每种检测方法获得的结果在重要方面不同：CFC检测方法的响应幅度始终较高，而CFC检测方法检测低水平响应的可能性低于ELISPOT检测方法。此外，ELISPOT和CFC结果之间缺乏数字一致性。对于需要检测低水平应答或将应答定义为阳性或阴性的研究，ELISPOT测定可能是首选。相比之下，CFC具有更大的动态范围，并允许响应细胞的表型区分，使其成为大多数其他应用的首选测定。(C)2003 Elsevier B. V.保留所有权利。
The enumeration of antigen-specific T cell responses has been greatly facilitated in recent years by the development of methods based on the detection of cytokines. In particular, the enzyme-linked immunospot (ELISPOT) and cytokine flow cytometry (CFC) assays have become popular. Since both assays are likely to continue to be in widespread use, it is important to evaluate whether their results are comparable. In the current study, we compared the results obtained in the ELISPOT and CFC assays using peptide pools corresponding to CMV and HIV-1 proteins in chronically HIV-1-infected individuals. Analysis of T cell responses to peptide pools indicated that the CMV pp65 and HIN-1 Gag CFC and ELISPOT-derived results were statistically correlated. However, the results obtained with each assay differed in important ways: the magnitude of the response was consistently higher in the CFC assay while the CFC assay was less likely than the ELISPOT assay to detect low-level responses. Furthermore, there was a lack of numeric agreement between ELISPOT and CFC results. For studies that require the detection of low-level responses, or definition of responses as positive or negative, the ELISPOT assay may be preferable. In contrast, the CFC has a greater dynamic range and allows for phenotypic discrimination of responding cells, making it the assay of choice for most other applications. (C) 2003 Elsevier B.V. All rights reserved.