Bruger Annika M, Vanhaver Christophe, Bruderek Kirsten, Amodio Giada, Tavukçuoğlu Ece, Esendagli Günes, Gregori Silvia, Brandau Sven, van der Bruggen Pierre
de Duve Institute, Université catholique de Louvain, Brussels, Belgium.
Research Division, Department of Otorhinolaryngology, University Hospital Essen, West German Cancer Center, Essen, Germany.
Methods Enzymol. 2020;632:155-192. doi: 10.1016/bs.mie.2019.05.046. Epub 2019 Jul 8.
Inhibition of T-cell proliferation is the most common approach to assess human myeloid-derived suppressor cell (MDSC) functions. However, diverse methodologies hinder the comparison of results obtained in different laboratories. In this chapter, we present a T-cell proliferation assay procedure based on allogeneic MDSC and T-cells that is potentially suitable to multi-center studies. The T-cells are isolated from non-cancerous donors and frozen for later use in different research groups. We observed that pure thawed T-cells showed poor proliferative capacities. To retain proliferation, T-cell-autologous mature dendritic cells are supplemented after thawing. MDSC are isolated from clinical samples and represent the sole variant between assays. Flow cytometry is used to assess T-cell proliferation by the dilution of a tracking dye.
抑制T细胞增殖是评估人髓源性抑制细胞（MDSC）功能最常用的方法。然而，多种方法阻碍了不同实验室所得结果的比较。在本章中，我们介绍一种基于同种异体MDSC和T细胞的T细胞增殖检测程序，该程序可能适用于多中心研究。T细胞从非癌症供体中分离出来并冷冻，供不同研究组日后使用。我们观察到，单纯解冻的T细胞增殖能力较差。为保持增殖能力，解冻后补充T细胞自体成熟树突状细胞。MDSC从临床样本中分离出来，是各检测之间唯一的变量。通过追踪染料的稀释，利用流式细胞术评估T细胞增殖。
