Functional analysis of highly defined, FACS-isolated populations of human regulatory CD4+CD25+ T cells
Functional analysis of highly defined, FACS-isolated populations of human regulatory CD4+CD25+ T cells
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DOI：
10.1016/j.clim.2005.02.018
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发表时间：
2005-04-01
影响因子：
8.6
通讯作者：
Hafler, DA
中科院分区：
文献类型：
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作者：
Baecher-Allan, C;Wolf, E;Hafler, DA
CD 4（+）CD 25（+）调节性T细胞（Treg）在维持免疫稳态中的重要性已经通过在小鼠中的许多自身免疫性疾病模型中对它们的操纵而在体内直接证明。在人调节细胞的研究中，我们发现，通过将CD 25（+）CD 4 T细胞的分离限制于仅表达最高水平的CD 25的那些细胞（约占总CD 4 T细胞的2-3%），可以最好地鉴定出始终显示出与体外鼠细胞所描述的最相似的体外调节活性的细胞。因此，CD 4（+）CD 25（高）亚群表现出与最初在小鼠中表征的CD 4（+）CD 25（+）调节细胞相同的体外特征。此外，表达中等至低水平的CD 25的细胞不仅不直接表现出离体抑制活性，而且实际上含有显著比例的CD 62 L（-）CD 4 T细胞，其被认为是体内活化的T细胞。由于使用CD 25作为Treg细胞纯化的标志物的固有困难，选择CD 4（+）CD 25（+）T细胞的CD 25高亚群使污染的活化CD 4 T细胞的共分离最小化的发现对于这些Treg细胞在人类疾病中的未来研究是重要的。为了进行这些研究，我们首先必须建立一个高度可重复的“微量体外共培养”测定系统，以使来自FACS分选的高纯度但低产量的调节群体的功能分析成为可能。有了这个系统，我们准备解剖高度特异性的CD 4（+）CD 25（+）细胞亚群所采用的机制的潜在异质性。(c)2005年爱思唯尔公司All rights reserved.
The importance of CD4(+)CD25(+) regulatory T cells (Treg) in maintaining immune homeostasis has been directly demonstrated in vivo by their manipulation in a number of autoimmune disease models in the mouse. In the study of human regulatory cells, we have found that the cells that consistently demonstrate the in vitro regulatory activity most similar to that described for murine cells in vitro are best identified by restricting the isolation of CD25(+)CD4 T cells to those cells expressing only the highest levels of CD25, representing approximately 2-3% of total CD4 T cells. Thus, it is the CD4(+)CD25(high) subset that exhibits the in vitro characteristics that are identical to the CD4(+)CD25(+) regulatory cells initially characterized in mice. Furthermore, the cells expressing medium to low levels of CD25 not only do not exhibit suppressive activity directly ex vivo, but also actually contain a significant proportion of CD62L(-)CD4 T cells which are believed to be in vivo activated T cells. Due to the inherent difficulties in using CD25 as a marker for the purification of Treg cells, the finding that selection of the CD25 high subset of CD4(+)CD25(+) T cells minimizes the co-isolation of contaminating activated CD4 T cells is important for future studies of these Treg cells in human disease. In order to perform these studies, we first had to establish a highly reproducible 'micro in vitro co-culture' assay system to enable the functional analysis of high-purity, but low-yield regulatory populations derived from FACS sorting. With this system in place, we are poised to dissect the potential heterogeneity of mechanisms employed by highly specific subpopulations of CD4(+)CD25(+) cells. (c) 2005 Elsevier Inc. All rights reserved.