Department of Immunology, University of Toronto, Toronto, Ontario M4N 3M5, Canada.
J Immunol. 2012 Oct 1;189(7):3411-20. doi: 10.4049/jimmunol.1200979. Epub 2012 Aug 27.
The generation of the cytotoxic CD8 T cell response is dependent on the functional outcomes imposed by the intrathymic constraints of differentiation and self-tolerance. Although thymic function can be partly replicated in vitro using OP9-DL1 cell cultures to yield CD8 αβ TCR-bearing cells from hematopoietic progenitor cells, a comprehensive and functional assessment of entirely in vitro generated CD8 T cells derived from bone marrow hematopoietic stem cells has not been established and remains controversial. In this study, we demonstrate that a phenotypic, molecular, and functional signature of in vitro derived CD8 T cells is akin to that of ex vivo CD8 T cells, although several significant differences were also observed. Transfer of in vitro derived CD8 T cells into syngeneic and immunodeficient host mice showed no graft-versus-host response, whereas a robust homeostatic proliferation was observed, respectively. These findings, along with a diverse and broad TCR repertoire expressed by the in vitro derived CD8 T cells, allowed for the successful generation of Ag-specific T cells to be obtained from an entirely in vitro generated CD8 T cell pool. These findings support the use of Ag-specific in vitro derived effector CD8 T cells for immune reconstitution approaches, which would be amenable to further tailoring for their use against viral infections or malignancies.
细胞毒性 CD8 T 细胞反应的产生依赖于分化和自身耐受的胸腺内限制所施加的功能结果。尽管使用 OP9-DL1 细胞培养物在体外部分复制了胸腺功能，从而从造血祖细胞中产生 CD8αβTCR 阳性细胞，但尚未建立和仍然存在争议的是，对完全来自骨髓造血干细胞的体外生成的 CD8 T 细胞进行全面和功能评估。在这项研究中，我们证明了体外生成的 CD8 T 细胞的表型、分子和功能特征类似于体外 CD8 T 细胞的特征，尽管也观察到了一些显著差异。体外生成的 CD8 T 细胞的转移到同基因和免疫缺陷宿主小鼠中未显示移植物抗宿主反应，而分别观察到了强烈的稳态增殖。这些发现，以及体外生成的 CD8 T 细胞表达的多样化和广泛的 TCR 谱，使得能够从完全体外生成的 CD8 T 细胞池中获得 Ag 特异性 T 细胞的成功生成。这些发现支持使用 Ag 特异性的体外衍生效应 CD8 T 细胞进行免疫重建方法，这将适合进一步针对它们用于对抗病毒感染或恶性肿瘤进行定制。
