Proliferation tracing with single-cell mass cytometry optimizes generation of stem cell memory-like T cells
Proliferation tracing with single-cell mass cytometry optimizes generation of stem cell memory-like T cells
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DOI：
10.1038/s41587-019-0033-2
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发表时间：
2019-03-01
影响因子：
46.9
通讯作者：
Bendall, Sean C.
中科院分区：
文献类型：
--
作者：
Good, Zinaida;Borges, Luciene;Bendall, Sean C.
将幼稚T细胞选择性分化为多能T细胞在临床上对于产生基于细胞的癌症免疫疗法具有很大的兴趣。细胞分化主要取决于分裂状态和分裂时间。在这里，我们采用了一种染料稀释法，通过大量细胞术和解偶联分裂，时间和调节蛋白表达来跟踪细胞增殖史，在复杂的体外环境中，单个幼稚的人T细胞在激活和扩增过程中。使用23个标记，我们定义了主要由分裂状态或时间控制的蛋白质组，发现未分裂的细胞占大部分表型多样性。接下来，我们构建了初始t细胞扩增过程中细胞状态变化的图谱。通过检查该图谱上的细胞信号传导，我们合理地选择了ibrutinib（一种BTK和ITK抑制剂），并在T细胞激活之前给药，以指导分化向T干细胞记忆（T- scm）样表型。这种跨越分裂状态和时间追踪细胞命运的方法可以广泛应用于指导细胞分化。
Selective differentiation of naive T cells into multipotent T cells is of great interest clinically for the generation of cell-based cancer immunotherapies. Cellular differentiation depends crucially on division state and time. Here we adapt a dye dilution assay for tracking cell proliferative history through mass cytometry and uncouple division, time and regulatory protein expression in single naive human T cells during their activation and expansion in a complex ex vivo milieu. Using 23 markers, we defined groups of proteins controlled predominantly by division state or time and found that undivided cells account for the majority of phenotypic diversity. We next built a map of cell state changes during naive T-cell expansion. By examining cell signaling on this map, we rationally selected ibrutinib, a BTK and ITK inhibitor, and administered it before T cell activation to direct differentiation toward a T stem cell memory (T-SCM)-like phenotype. This method for tracing cell fate across division states and time can be broadly applied for directing cellular differentiation.