Duggleby Richard C, Shaw Tovah N F, Jarvis Lorna B, Kaur Gurman, Gaston J S Hill
Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
Immunology. 2007 May;121(1):129-39. doi: 10.1111/j.1365-2567.2006.02550.x.
It is clear that regulatory T cells (Treg) have an important role in preventing autoimmunity and modulating responses to pathogens. Full characterization of Treg cell function in human patients would be greatly facilitated by practical methods for expanding Treg in vitro. Methods for expansion have been reported but whether expression of surface and intracellular markers associated with freshly isolated Treg following expansion correlates with the maintenance of function is unclear. Our aim was to investigate the various methods of expansion and to correlate regulatory activity with expression of these markers. We show that, of the markers associated with freshly isolated Treg, only CD27 expression correlated with regulatory activity and could be used to isolate cells with regulatory activity from lines expanded from CD4+ CD25+ cells. Also, cells expressing high levels of the transcription factor forkhead box P3 (Foxp3) were confined to the CD27+ population within these lines. Expression of CD27 by cells in lines expanded from CD4+ CD25- cells varied depending on the stimulus used for expansion, but these lines did not have significant regulatory activity even when the CD27+ cells were tested. Analysis of synovial CD4+ CD25+ cells from reactive arthritis patients revealed that they were predominantly CD27 positive. This also applied to CD25(high) and CD25(intermediate) CD4+ cells, despite their reported different abilities to regulate. We conclude that, whilst CD27 is useful for identifying Treg in the cell lines obtained after expansion of CD4+ CD25+ cells, its expression may not reliably identify the Treg cell population in other T-cell populations such as those found in joints.
很明显，调节性T细胞（Treg）在预防自身免疫和调节对病原体的反应中发挥着重要作用。体外扩增Treg的实用方法将极大地促进对人类患者Treg细胞功能的全面表征。已经报道了扩增方法，但扩增后与新鲜分离的Treg相关的表面和细胞内标志物的表达是否与功能维持相关尚不清楚。我们的目的是研究各种扩增方法，并将调节活性与这些标志物的表达相关联。我们发现，在与新鲜分离的Treg相关的标志物中，只有CD27表达与调节活性相关，并且可用于从CD4 + CD25 +细胞扩增的细胞系中分离具有调节活性的细胞。此外，在这些细胞系中，高水平表达转录因子叉头框P3（Foxp3）的细胞局限于CD27 +群体。从CD4 + CD25-细胞扩增的细胞系中细胞的CD27表达因用于扩增的刺激而异，但即使对CD27 +细胞进行测试，这些细胞系也没有显著的调节活性。对反应性关节炎患者滑膜CD4 + CD25 +细胞的分析显示，它们主要为CD27阳性。这也适用于CD25（高）和CD25（中）CD4 +细胞，尽管它们报道的调节能力不同。我们得出结论，虽然CD27有助于识别CD4 + CD25 +细胞扩增后获得的细胞系中的Treg，但其表达可能无法可靠地识别其他T细胞群体（如关节中发现的那些）中的Treg细胞群体。
