A Converse 4-1BB and CD40 Ligand Expression Pattern Delineates Activated Regulatory T Cells (Treg) and Conventional T Cells Enabling Direct Isolation of Alloantigen-Reactive Natural Foxp3+ Treg
A Converse 4-1BB and CD40 Ligand Expression Pattern Delineates Activated Regulatory T Cells (Treg) and Conventional T Cells Enabling Direct Isolation of Alloantigen-Reactive Natural Foxp3+ Treg
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DOI：
10.4049/jimmunol.1201090
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发表时间：
2012-12-15
影响因子：
4.4
通讯作者：
Thiel, Andreas
中科院分区：
文献类型：
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作者：
Schoenbrunn, Anne;Frentsch, Marco;Thiel, Andreas
天然调节性 T 细胞 (nTreg) 在免疫耐受的诱导和维持中发挥着核心作用。实验移植模型和最近的临床试验表明 nTreg 可以控制同种异体反应性。为了将基于 Treg 的细胞疗法升级为选择性抑制不良免疫反应，只有 Ag 特异性 nTreg 的转移才是合适的治疗选择。然而，Ag 特异性 nTreg 在外周以极低的频率存在，并且迄今为止缺少用于精确识别的适当表面标记。在这项研究中，我们证明激活的 nTreg 细胞和激活的常规 T 细胞的 4-1BB 和 CD40 配体 (CD40L) 表达特征不同，从而可以清楚地区分彼此。基于4-1BB的表达和CD40L表达的缺失，可以直接从MLR培养物中高纯度地分离出人同种异体抗原反应性Foxp3(+) nTreg。同种抗原反应性4-1BB(+)CD40L(-) nTreg的特征是完全去甲基化的Treg特异性去甲基化区域，并且表现出优于多克隆Treg的同种抗原特异性抑制特性。重要的是，分离的4-1BB(+)CD40L(-) nTreg 在体外扩增后仍保持nTreg 表型和同种异体抗原反应性。我们的结果提供了同时分析 Ag 特异性 nTreg 和常规 T 细胞的可能性，并利用 Ag 特异性 nTreg 建立细胞疗法，旨在特异性抑制不需要的免疫。免疫学杂志，2012，189：5985-5994。
Natural regulatory T cells (nTreg) play a central role in the induction and maintenance of immunological tolerance. Experimental transplant models and recent clinical trials demonstrate that nTreg can control alloreactivity. To upgrade Treg-based cell therapies to a selective suppression of undesired immune reactions, only the transfer of Ag-specific nTreg represents the appropriate therapeutic option. However, Ag-specific nTreg are present at extremely low frequencies in the periphery, and so far appropriate surface markers for their precise identification are missing. In this study, we demonstrate that activated nTreg and activated conventional T cells differ in their 4-1BB and CD40 ligand (CD40L) expression signatures, allowing a clear dissection from each other. Based on the expression of 4-1BB and absence of CD40L expression, human alloantigen-reactive Foxp3(+) nTreg can be directly isolated from MLR cultures with high purity. Alloantigen-reactive 4-1BB(+)CD40L(-) nTreg were characterized by a completely demethylated Treg-specific demethylated region and showed alloantigen-specific suppressive properties superior to polyclonal Treg. Importantly, isolated 4-1BB(+)CD40L(-) nTreg maintain the nTreg phenotype and alloantigen-reactivity after in vitro expansion. Our results offer the possibility to simultaneously analyze Ag-specific nTreg and conventional T cells, and to establish cellular therapies with Ag-specific nTreg aiming at a specific inhibition of unwanted immunity. The Journal of Immunology, 2012, 189: 5985-5994.