Molecular Immunology Laboratory, Women's and Children's Health Research Institute, Women's and Children's Hospital, North Adelaide, South Australia, Australia.
Immunol Cell Biol. 2012 Sep;90(8):812-21. doi: 10.1038/icb.2012.18. Epub 2012 Jun 12.
Natural Regulatory T cells (Tregs) are defined by stable expression of the cell surface proteins CD4 and CD25, low surface expression of CD127 and expression of the transcription factor FOXP3. The contribution of Treg to the prevention of autoimmunity and the maintenance of immune homoestasis is the subject of ongoing interest, as alterations in Treg numbers and function are implicated in a wide range of diseases. The in vitro benchmark for determining Treg function is suppression of proliferation of unmatched effector T cells in a mixed lymphocyte reaction (MLR) over a 3-6-day time period. As an alternative to this assay, we show that a 7-h CD154 expression assay is rapid, simple and provides a reliable readout of suppressor function. Using multiple Treg-like cell types including natural (n)Treg, inducible (i)Treg and Treg cell lines, we show that suppression of CD154 expression is a surrogate for suppression of proliferation. We propose this as a suitable alternative to the MLR assay, as it is rapid and may be more amenable to high-throughput screening, analysing large cohorts of clinical samples or assaying transiently suppressive populations.
天然调节性 T 细胞（Tregs）的特征是稳定表达细胞表面蛋白 CD4 和 CD25，低表面表达 CD127 和转录因子 FOXP3。Treg 对预防自身免疫和维持免疫内稳的贡献是目前研究的热点，因为 Treg 数量和功能的改变与多种疾病有关。体外测定 Treg 功能的基准是在混合淋巴细胞反应（MLR）中抑制匹配效应 T 细胞在 3-6 天时间内的增殖。作为该测定的替代方法，我们表明，7 小时 CD154 表达测定快速、简单，并提供了抑制功能的可靠读数。使用包括天然（n）Treg、诱导（i）Treg 和 Treg 细胞系在内的多种 Treg 样细胞类型，我们表明 CD154 表达的抑制是增殖抑制的替代物。我们提出这是 MLR 测定的合适替代方法，因为它快速，可能更适合高通量筛选、分析大量临床样本或测定瞬时抑制群体。
