Long Anna E, Tatum Megan, Mikacenic Carmen, Buckner Jane H
Translational Research Program, Benaroya Research Institute at Virginia Mason, Seattle, WA, USA.
Department of Medicine, University of Washington, Seattle, WA, USA.
J Immunol Methods. 2017 Oct;449:15-22. doi: 10.1016/j.jim.2017.06.009. Epub 2017 Jun 22.
Measuring regulatory T cell suppression provides important insight into T cell dysfunction in autoimmune disease. However, to date, suppression assays are limited by the requirement for freshly isolated cells, and significant cell numbers. Here, we present a novel and rapid in vitro assay using effector T cell surface expression of both CD25 and CD134 as a surrogate marker of regulatory T cell-mediated suppression. This surface marker-based suppression assay works for frozen samples and for samples with limited cell numbers. It is also shorter taking two days to complete compared to the four days required for proliferation-based assays. Furthermore, this assay works with both in vitro expanded and natural Tregs, as well as anti-CD3/anti-CD28 bead-based and APC stimulation conditions. In conclusion, we have developed and validated a new suppression assay for cryopreserved samples with limited cell numbers that may be helpful to investigate T cell regulation in the context of infection or autoimmune diseases.
测量调节性T细胞抑制作用可为深入了解自身免疫性疾病中的T细胞功能障碍提供重要线索。然而，迄今为止，抑制试验受到新鲜分离细胞的需求以及大量细胞数量的限制。在此，我们提出了一种新型快速体外试验，利用效应T细胞表面表达的CD25和CD134作为调节性T细胞介导抑制作用的替代标志物。这种基于表面标志物的抑制试验适用于冷冻样本和细胞数量有限的样本。与基于增殖的试验所需的四天相比，它完成时间更短，只需两天。此外，该试验适用于体外扩增的天然调节性T细胞，以及基于抗CD3/抗CD28磁珠和抗原呈递细胞刺激的条件。总之，我们已经开发并验证了一种针对细胞数量有限的冷冻样本的新型抑制试验，这可能有助于在感染或自身免疫性疾病背景下研究T细胞调节。
