Institute for Medical Immunology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
Berlin Center for Advanced Therapies (BeCAT), Charité-Universitätsmedizin Berlin, Berlin, Germany.
Front Immunol. 2019 Feb 5;10:150. doi: 10.3389/fimmu.2019.00150. eCollection 2019.
CD4CD25FoxP3 human regulatory T (T) are promising candidates for reshaping undesired immunity/inflammation by adoptive cell transfer, yet their application is strongly dependent on robust assays testing their functionality. Several studies along with first clinical data indicate T to be auspicious to use for future cell therapies, e.g., to induce tolerance after solid organ transplantation. To this end, T suppressive capacity has to be thoroughly evaluated prior to any therapeutic application. A 7 h-protocol for the assessment of T function by suppression of the early activation markers CD154 and CD69 on CD4CD25 responder T (T) upon polyclonal stimulation via αCD3/28-coated activating microbeads has previously been published. Even though this assay has since been applied by various groups, the protocol comes with a critical pitfall, which is yet not corrected by the journal of its original publication. Our results demonstrate that the observed decrease in activation marker frequency on T is due to competition for αCD3/28-coated microbeads as opposed to a T-attributable effect and therefore the protocol cannot further be used as a diagnostic test to assess suppressive T function.
CD4CD25FoxP3 人类调节性 T（T）细胞是通过过继细胞转移重塑不良免疫/炎症的有前途的候选者，但它们的应用强烈依赖于测试其功能的强大检测方法。多项研究和初步临床数据表明，T 细胞有望用于未来的细胞治疗，例如，在实体器官移植后诱导耐受。为此，在进行任何治疗性应用之前，必须彻底评估 T 的抑制能力。先前已经发表了一种 7 小时方案，用于通过αCD3/28 包被的激活微珠对多克隆刺激后的 CD4CD25 应答 T（T）细胞上的早期激活标志物 CD154 和 CD69 的抑制来评估 T 细胞的功能。尽管该检测方法已被不同的研究小组应用，但该方案存在一个关键缺陷，原始出版物的期刊尚未对此进行纠正。我们的结果表明，T 细胞上观察到的激活标志物频率下降是由于对αCD3/28 包被的微珠的竞争所致，而不是 T 细胞归因的效应，因此该方案不能再用作评估抑制性 T 细胞功能的诊断测试。
