Gregori Silvia, Bacchetta Rosa, Passerini Laura, Levings Megan K, Roncarolo Maria Grazia
San Raffaele Telethon Institute for Gene Therapy, Milan, Italy.
Methods Mol Biol. 2007;380:83-105. doi: 10.1007/978-1-59745-395-0_6.
Regulatory T cells play a central role in controlling homeostasis, and in inducing and maintaining tolerance to both foreign and self-antigens. Several types of T cells with regulatory activity have been described both in mice and humans, and those within the CD4+ subset have been extensively studied. Among them, the best characterized are the naturally occurring CD4+CD25+ regulatory T (Treg) cells, and the adaptive type 1 regulatory T (Tr1) cells. Natural Treg cells can arise directly from the thymus, are characterized by the constitutive expression of the transcription factor Foxp3, and suppress T cell responses in a cell-cell contact mediated mechanism. On the contrary, adaptive Tr1 cells arise in the periphery upon encountering antigen in a tolerogenic environment, produce high levels of interleukin (IL)-10 and mediate suppression via IL-10. During the last decade, much effort has been placed on developing protocols to generate regulatory T-cell lines and clones, to further define the similarities and differences between various regulatory T-cell subsets. In this chapter, we will outline protocols to expand naturally occurring Treg cells, to differentiate homogeneous population of Tr1 cells in vitro, and to generate natural Treg and Tr1 cell clones and cell lines.
调节性T细胞在控制体内平衡以及诱导和维持对外源和自身抗原的耐受性方面发挥着核心作用。在小鼠和人类中均已描述了几种具有调节活性的T细胞类型，其中CD4+亚群中的T细胞已得到广泛研究。其中，特征最明确的是天然存在的CD4+CD25+调节性T（Treg）细胞和适应性1型调节性T（Tr1）细胞。天然Treg细胞可直接源自胸腺，其特征在于转录因子Foxp3的组成性表达，并通过细胞间接触介导的机制抑制T细胞反应。相反，适应性Tr1细胞在致耐受性环境中遇到抗原后在外周产生，产生高水平的白细胞介素（IL）-10，并通过IL-10介导抑制作用。在过去十年中，人们付出了很多努力来开发生成调节性T细胞系和克隆的方案，以进一步明确各种调节性T细胞亚群之间的异同。在本章中，我们将概述扩增天然存在的Treg细胞、在体外分化出均一的Tr1细胞群体以及生成天然Treg和Tr1细胞克隆及细胞系的方案。
