Activation-induced CD137 is a fast assay for identification and multi-parameter flow cytometric analysis of alloreactive T cells
Activation-induced CD137 is a fast assay for identification and multi-parameter flow cytometric analysis of alloreactive T cells
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DOI：
10.1111/cei.12152
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发表时间：
2013-10-01
影响因子：
4.6
通讯作者：
Betjes, M. G. H.
中科院分区：
文献类型：
--
作者：
Litjens, N. H. R.;de Wit, E. A.;Betjes, M. G. H.
在单细胞水平上检测和分离活的同种异体反应性 T 细胞需要在 T 细胞受体激活时特异性诱导的细胞表面标记。在这项研究中，研究了肿瘤坏死因子受体 (TNFR) 家族成员 CD137 (4-1BB) 识别循环同种异体 T 细胞总数的潜力。建立了敏感和特异性检测循环 T 细胞上同种异体诱导的 CD137 表达的最佳条件。此后，通过多参数流式细胞术对CD137(+)同种异体反应性T细胞进行表型和功能表征。同种抗原诱导的CD137表达鉴定出同种异体反应性CD8(+) T细胞(平均值+/-平均值标准误差：0.21 +/- 0.07%)和同种异体反应性CD4(+) T细胞(0.21 +/- 0.05%)。 CD137(+)同种异体反应性T细胞在不同的T细胞亚群中检测到，包括初始T细胞，但优先在CD28(+) T细胞中发现，而不是在终末分化T细胞亚群中发现。在同种异体（再）刺激下，T 细胞的细胞因子产生和增殖能力主要位于 CD137 表达部分。大约10%的CD137(+)同种反应性T细胞产生干扰素(IFN)-γ、白细胞介素(IL)-2和TNF-α的任意组合。由多种细胞因子表达定义的多功能同种反应性 T 细胞很少被观察到。总之，激活诱导的 CD137 表达是一种快速检测方法，允许通过多参数流式细胞术在单细胞水平上对总同种异体反应性 T 细胞区室进行检测和功能分析。
Detection and isolation of viable alloreactive T cells at the single-cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)-family, CD137 (4-1BB) was investigated for its potential to identify the total pool of circulating alloreactive T cells. Optimal conditions for sensitive and specific detection of allogeneic-induced CD137 expression on circulating T cells were established. Thereafter, CD137(+) alloreactive T cells were phenotypically and functionally characterized by multi-parameter flow cytometry. Alloantigen-induced CD137 expression identified both alloreactive CD8(+) T cells (mean +/- standard error of the mean: 0.21 +/- 0.07%) and alloreactive CD4(+) T cells (0.21 +/- 0.05%). CD137(+) alloreactive T cells were detected in different T cell subsets, including naive T cells, but were found preferentially in CD28(+) T cells and not in the terminally differentiated T cell subset. Upon allogeneic (re-)stimulation, the cytokine-producing as well as proliferative capacity of T cells resided mainly within the CD137-expressing fraction. About 10% of the CD137(+) alloreactive T cells produced any combination of interferon (IFN)-gamma, interleukin (IL)-2 and TNF-alpha. Polyfunctional alloreactive T cells, defined by multiple cytokine expression, were observed infrequently. In conclusion, activation-induced CD137 expression is a fast assay allowing for detection and functional analysis of the total alloreactive T cell compartment at the single-cell level by multi-parameter flow cytometry.