San Raffaele Telethon Institute for Gene Therapy, Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan, Italy.
Mol Ther. 2012 Sep;20(9):1778-1790. doi: 10.1038/mt.2012.71. Epub 2012 Jun 12.
Type 1 regulatory T (Tr1) cells are an inducible subset of CD4(+) Tr cells characterized by high levels of interleukin (IL)-10 production and regulatory properties. Several protocols to generate human Tr1 cells have been developed in vitro. However, the resulting population includes a significant fraction of contaminating non-Tr1 cells, representing a major bottleneck for clinical application of Tr1 cell therapy. We generated an homogeneous IL-10-producing Tr1 cell population by transducing human CD4(+) T cells with a bidirectional lentiviral vector (LV) encoding for human IL-10 and the marker gene, green fluorescent protein (GFP), which are independently coexpressed. The resulting GFP(+) LV-IL-10-transduced human CD4(+) T (CD4(LV-IL-10)) cells expressed, upon T-cell receptor (TCR) activation, high levels of IL-10 and concomitant low levels of IL-4, and markers associated with IL-10. Moreover, CD4(LV-IL-10) T cells displayed typical Tr1 features: the anergic phenotype, the IL-10, and transforming growth factor (TGF)-β dependent suppression of allogeneic T-cell responses, and the ability to suppress in a cell-to-cell contact independent manner in vitro. CD4(LV-IL-10) T cells were able to control xeno graft-versus-host disease (GvHD), demonstrating their suppressive function in vivo. These results show that constitutive over-expression of IL-10 in human CD4(+) T cells leads to a stable cell population that recapitulates the phenotype and function of Tr1 cells.
1 型调节性 T（Tr1）细胞是 CD4（+）Tr 细胞的一个诱导亚群，其特征是高水平的白细胞介素（IL）-10 产生和调节特性。已经开发了几种体外生成人 Tr1 细胞的方案。然而，由此产生的群体包括大量污染的非 Tr1 细胞，这是 Tr1 细胞治疗临床应用的主要瓶颈。我们通过转导带有双向慢病毒载体（LV）的人 CD4（+）T 细胞生成了一种同质的 IL-10 产生 Tr1 细胞群体，该载体编码人 IL-10 和标记基因绿色荧光蛋白（GFP），它们独立共表达。所得的 GFP（+）LV-IL-10 转导的人 CD4（+）T（CD4（LV-IL-10））细胞在 T 细胞受体（TCR）激活时表达高水平的 IL-10 和伴随的低水平的 IL-4，以及与 IL-10 相关的标记物。此外，CD4（LV-IL-10）T 细胞表现出典型的 Tr1 特征：无反应性表型、IL-10 和转化生长因子（TGF）-β依赖性抑制同种异体 T 细胞反应，以及在体外以非细胞接触方式抑制的能力。CD4（LV-IL-10）T 细胞能够控制异种移植物抗宿主病（GvHD），证明了它们在体内的抑制功能。这些结果表明，在人 CD4（+）T 细胞中持续过表达 IL-10 可导致稳定的细胞群体，该群体再现了 Tr1 细胞的表型和功能。
