Induction of antigen-specific regulatory T cells (Tregs) in vivo is the holy grail of current immune-regulating therapies in autoimmune diseases, such as type 1 diabetes. Tolerogenic dendritic cells (tolDCs) generated from monocytes by a combined treatment with vitamin D and dexamethasone (marked by CD52hi and CD86lo expression) induce antigen-specific Tregs. We evaluated the phenotypes of these Tregs using high-dimensional mass cytometry to identify a surface-based T cell signature of tolerogenic modulation. Naïve CD4+ T cells were stimulated with tolDCs or mature inflammatory DCs pulsed with proinsulin peptide, after which the suppressive capacity, cytokine production and phenotype of stimulated T cells were analysed. TolDCs induced suppressive T cell lines that were dominated by a naïve phenotype (CD45RA+CCR7+). These naïve T cells, however, did not show suppressive capacity, but were arrested in their naïve status. T cell cultures stimulated by tolDC further contained memory-like (CD45RA-CCR7-) T cells expressing regulatory markers Lag-3, CD161 and ICOS. T cells expressing CD25lo or CD25hi were most prominent and suppressed CD4+ proliferation, while CD25hi Tregs also effectively supressed effector CD8+ T cells. We conclude that tolDCs induce antigen-specific Tregs with various phenotypes. This extends our earlier findings pointing to a functionally diverse pool of antigen-induced and specific Tregs and provides the basis for immune-monitoring in clinical trials with tolDC.

中文翻译：

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耐受性树突状细胞诱导的胰岛素原肽特异性调节性T细胞的多维分析。

体内抗原特异性调节性T细胞（Tregs）的诱导是目前自身免疫性疾病（例如1型糖尿病）中免疫调节疗法的圣杯。通过用维生素D和地塞米松（以CD52hi和CD86lo表达为标志）联合处理，从单核细胞产生的耐受性树突状细胞（tolDC）诱导抗原特异性Treg。我们使用高维质量流式细胞仪评估这些Treg的表型，以鉴定致耐受性调节的基于表面的T细胞标志。用tolDCs或用胰岛素原肽脉冲的成熟炎性DCs刺激幼稚的CD4 + T细胞，然后分析刺激的T细胞的抑制能力，细胞因子产生和表型。TolDCs诱导的抑制性T细胞系以幼稚的表型（CD45RA + CCR7 +）为主。但是，这些幼稚的T细胞 没有表现出抑制能力，但以幼稚的身份被捕。受tolDC刺激的T细胞培养物进一步包含表达调节标记Lag-3，CD161和ICOS的记忆样（CD45RA-CCR7-）T细胞。表达CD25lo或CD25hi的T细胞最为突出，并抑制了CD4 +增殖，而CD25hi Tregs也有效地抑制了效应CD8 + T细胞。我们得出的结论是，tolDCs诱导具有各种表型的抗原特异性Treg。这扩展了我们先前的发现，指向功能多样的抗原诱导的和特定的Treg，并为tolDC的临床试验提供了免疫监测的基础。表达CD25lo或CD25hi的T细胞最为突出，并抑制了CD4 +增殖，而CD25hi Tregs也有效地抑制了效应CD8 + T细胞。我们得出的结论是，tolDCs诱导具有各种表型的抗原特异性Treg。这扩展了我们先前的发现，指向功能多样的抗原诱导的和特定的Treg，并为tolDC的临床试验提供了免疫监测的基础。表达CD25lo或CD25hi的T细胞最为突出，并抑制了CD4 +增殖，而CD25hi Tregs也有效地抑制了效应CD8 + T细胞。我们得出的结论是，tolDCs诱导具有各种表型的抗原特异性Treg。这扩展了我们先前的发现，指向功能多样的抗原诱导的和特定的Treg，并为tolDC的临床试验提供了免疫监测的基础。