Miyara Makoto, Chader Driss, Burlion Aude, Goldstein Jérémie, Sterlin Delphine, Norol Françoise, Trebeden-Nègre Hélène, Claër Laetitia, Sakaguchi Shimon, Marodon Gilles, Amoura Zahir, Gorochov Guy
Department of immunology, AP-HP Pitié Salpêtrière, Paris, France.
Sorbonne Universités, UPMC Univ Paris 06, INSERM, CNRS, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), Paris, France.
Oncotarget. 2016 Jul 28;8(62):104733-104744. doi: 10.18632/oncotarget.10914. eCollection 2017 Dec 1.
FOXP3 regulatory T cell (Treg) based cellular therapies represent promising therapeutic options in autoimmunity, allergy, transplantation and prevention of Graft Versus Host (GVH) Disease. Among human FOXP3-expressing CD4T cells, only the CD45RA naïve Treg (nTreg) subset is suitable for expansion. However, FoxP3 expression decays in cells using currently described culture protocols. Rapamycin alone was not able to prevent FOXP3 loss in nTregs cells, as only a half of them maintained FOXP3 expression after 14 days of culture. In contrast we report a novel combined drug regimen that can drastically stabilize FOXP3 expression in cultured Tregs. IL-2, rapamycin, histone deacetylase and DNA methyltransferase inhibitors act in synergy to allow expansion of human regulatory T cells with sustained high expression of FOXP3 and CD15s with potent suppressive capacities and control of murine xeno-GVH reactions. Of note, an additional subsequent infusion of expanded nTreg cells did not improve survival of mice. Combination of IL-2, rapamycin, histone deacetylase and DNA methyltransferase inhibitors is optimal for the expansion of pure effective nTreg maintaining high levels of FOXP3 for therapeutic purposes.
基于叉头状转录因子3（FOXP3）调节性T细胞（Treg）的细胞疗法在自身免疫性疾病、过敏、移植以及预防移植物抗宿主病（GVHD）方面展现出了极具前景的治疗选择。在人类表达FOXP3的CD4T细胞中，只有CD45RA初始Treg（nTreg）亚群适合扩增。然而，按照目前所描述的培养方案，细胞中的FoxP3表达会逐渐衰减。单独使用雷帕霉素无法阻止nTreg细胞中FOXP3的丢失，因为在培养14天后，只有一半的细胞仍维持FOXP3表达。相比之下，我们报告了一种新型联合用药方案，该方案能够显著稳定培养的Treg细胞中的FOXP3表达。白细胞介素-2（IL-2）、雷帕霉素、组蛋白去乙酰化酶和DNA甲基转移酶抑制剂协同作用，可使人调节性T细胞得以扩增，同时维持FOXP3和CD15s的持续高表达，并具备强大的抑制能力，还能控制小鼠异种GVHD反应。值得注意的是，额外输注扩增后的nTreg细胞并不能提高小鼠的存活率。IL-2、雷帕霉素、组蛋白去乙酰化酶和DNA甲基转移酶抑制剂的联合使用对于扩增纯有效的nTreg最为理想，这种nTreg能够维持高水平的FOXP3，可用于治疗目的。
