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Activation and expansion of human T cells using artificial antigen-presenting cell scaffolds
Nature Protocols ( IF 14.8 ) Pub Date : 2020-01-13 , DOI: 10.1038/s41596-019-0249-0
David K Y Zhang 1, 2 , Alexander S Cheung 3 , David J Mooney 1, 2
Affiliation  

Synthetic antigen-presenting cells (APCs) are used to mediate scalable ex vivo T-cell expansion for adoptive cell therapy. Recently, we developed APC-mimetic scaffolds (APC-ms), which present signals to T cells in a physiological manner to mediate rapid and controlled T-cell expansion. APC-ms are composed of individual high-aspect-ratio silica microrods loaded with soluble mitogenic cues and coated with liposomes of defined compositions, to form supported lipid bilayers. Membrane-bound ligands for stimulation and co-stimulation of T-cell receptors are presented via the fluid, synthetic membranes, while mitogenic cues are released slowly from the microrods. In culture, interacting T cells assemble the individual APC-ms microrods into a biodegradable 3D matrix. Compared to conventional methods, APC-ms facilitates several-fold greater polyclonal T-cell expansion and improved antigen-specific enrichment of rare T-cell subpopulations. Here we provide a detailed protocol for APC-ms synthesis and use for human T-cell activation, and discuss important considerations for material design and T-cell co-culture. This protocol describes the facile assembly of APC-ms in ~4 h and rapid expansion or enrichment of relevant T-cell clones in <2 weeks, and is applicable for T-cell manufacturing and assay development.



中文翻译:

使用人工抗原呈递细胞支架激活和扩增人类 T 细胞

合成抗原呈递细胞 (APC) 用于介导可扩展的体外 T 细胞扩增,用于过继细胞治疗。最近,我们开发了 APC 模拟支架 (APC-ms),它以生理方式向 T 细胞提供信号,以介导快速和受控的 T 细胞扩增。APC-ms 由单独的高纵横比二氧化硅微棒组成,这些微棒装载有可溶性有丝分裂线索,并涂有特定成分的脂质体,以形成支持的脂质双层。用于刺激和共刺激 T 细胞受体的膜结合配体通过流体合成膜呈现,而有丝分裂线索从微棒中缓慢释放。在培养中,相互作用的 T 细胞将单个 APC-ms 微棒组装成可生物降解的 3D 基质。与传统方法相比,APC-ms 促进多克隆 T 细胞扩增数倍,并改善稀有 T 细胞亚群的抗原特异性富集。在这里,我们提供了 APC-ms 合成和用于人类 T 细胞活化的详细协议,并讨论了材料设计和 T 细胞共培养的重要考虑因素。该协议描述了在 4 小时内轻松组装 APC-ms 并在 <2 周内快速扩展或富集相关 T 细胞克隆,适用于 T 细胞制造和检测开发。

更新日期:2020-01-13
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