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Abstract：
Regulatory T cell (Treg)-mediated immunosuppression is considered a major obstacle for successful cancer immunotherapy. The association between clinical outcome and Tregs is being studied extensively in clinical trials, but unfortunately, no consensus has been reached about (a) the markers and (b) the gating strategy required to define human Tregs in this context, making it difficult to draw final conclusions. Therefore, we have organized an international workshop on the detection and functional testing of Tregs with leading experts in the field, and 40 participants discussing different analyses and the importance of different markers and context in which Tregs were analyzed. This resulted in a rationally composed ranking list of \"Treg markers\". Subsequently, the proposed Treg markers were tested to get insight into the overlap/differences between the most frequently used Treg definitions and their utility for Treg detection in various human tissues. Here, we conclude that the CD3, CD4, CD25, CD127, and FoxP3 markers are the minimally required markers to define human Treg cells. Staining for Ki67 and CD45RA showed to provide additional information on the activation status of Tregs. The use of markers was validated in a series of PBMC from healthy donors and cancer patients, as well as in tumor-draining lymph nodes and freshly isolated tumors. In conclusion, we propose an essential marker set comprising antibodies to CD3, CD4, CD25, CD127, Foxp3, Ki67, and CD45RA and a corresponding robust gating strategy for the context-dependent analysis of Tregs by flow cytometry.
摘要：
调节性T细胞（Treg）介导的免疫抑制被认为是成功的癌症免疫疗法的主要障碍。临床试验中正在广泛研究临床结果与Tregs之间的关联，但不幸的是,关于（a）标记和（b）在这种情况下定义人类Treg所需的门控策略尚未达成共识，很难得出最后的结论。因此,我们与该领域的领先专家组织了一次关于Tregs检测和功能测试的国际研讨会，40名参与者讨论了不同的分析以及不同标记的重要性以及分析Tregs的背景。这导致了“Treg标记”的合理组成的排名列表。随后，对所提出的Treg标记进行了测试,以深入了解最常用的Treg定义之间的重叠/差异及其在各种人体组织中检测Treg的实用性.这里,我们得出的结论是,CD3,CD4,CD25,CD127和FoxP3标志物是定义人Treg细胞所需的最低限度标志物.Ki67和CD45RA的染色显示提供了有关Tregs激活状态的其他信息。在一系列健康供体和癌症患者的PBMC中验证了标记物的使用，以及肿瘤引流淋巴结和新鲜分离的肿瘤。总之,我们提出了一个包含针对CD3,CD4,CD25,CD127,Foxp3,Ki67和CD45RA的抗体的基本标志物集,以及一个相应的稳健门控策略,用于通过流式细胞术对Tregs进行上下文相关分析.
