Huang Qing, Lam Avery J, Boardman Dominic A, Dawson Nicholas A J, Levings Megan K
Department of Surgery, University of British Columbia, Vancouver, Canada.
BC Children's Hospital Research Institute, Vancouver, Canada.
Bio Protoc. 2021 Nov 5;11(21):e4217. doi: 10.21769/BioProtoc.4217.
Regulatory T cells (Tregs) suppress immune responses via a variety of mechanisms and can be used as a cellular therapy to induce tolerance. The function of Tregs is commonly assessed in vitro using assays that measure suppression of effector T cell proliferation and/or cytokine production. However, Tregs can also suppress the function of antigen presenting cells, creating a need for methodology to routinely measure this aspect of their function. This protocol describes a method to measure human Treg-mediated suppression of CD80 and CD86 expression on mature, monocyte-derived dendritic cells. Representative data show suppression mediated by polyclonal Tregs as well as antigen-specific Tregs generated using chimeric antigen receptor (CAR) technology. This method can be used in parallel to T cell suppression assays to measure the functional activity of human Tregs.
调节性T细胞（Tregs）通过多种机制抑制免疫反应，可作为一种细胞疗法来诱导免疫耐受。Tregs的功能通常在体外通过检测效应T细胞增殖抑制和/或细胞因子产生的试验来评估。然而，Tregs也能抑制抗原呈递细胞的功能，因此需要一种方法来常规测量其这方面的功能。本方案描述了一种测量人Tregs介导的对成熟单核细胞来源的树突状细胞上CD80和CD86表达抑制作用的方法。代表性数据显示了多克隆Tregs以及使用嵌合抗原受体（CAR）技术产生的抗原特异性Tregs介导的抑制作用。该方法可与T细胞抑制试验并行使用，以测量人Tregs的功能活性。
