The production of autologous T cells expressing a chimaeric antigen receptor (CAR) is time-consuming, costly and occasionally unsuccessful. T-cell-derived induced pluripotent stem cells (TiPS) are a promising source for the generation of ‘off-the-shelf’ CAR T cells, but the in vitro differentiation of TiPS often yields T cells with suboptimal features. Here we show that the premature expression of the T-cell receptor (TCR) or a constitutively expressed CAR in TiPS promotes the acquisition of an innate phenotype, which can be averted by disabling the TCR and relying on the CAR to drive differentiation. Delaying CAR expression and calibrating its signalling strength in TiPS enabled the generation of human TCR^– CD8αβ⁺ CAR T cells that perform similarly to CD8αβ⁺ CAR T cells from peripheral blood, achieving effective tumour control on systemic administration in a mouse model of leukaemia and without causing graft-versus-host disease. Driving T-cell maturation in TiPS in the absence of a TCR by taking advantage of a CAR may facilitate the large-scale development of potent allogeneic CD8αβ⁺ T cells for a broad range of immunotherapies.

表达嵌合抗原受体 (CAR) 的自体 T 细胞的生产既费时又昂贵，而且有时会失败。T 细胞衍生的诱导多能干细胞 (TiPS) 是产生“现成”CAR T 细胞的有前途的来源，但 TiPS 的体外分化通常会产生具有次优特征的 T 细胞。在这里，我们表明 T 细胞受体 (TCR) 或 TiPS 中组成型表达的 CAR 的过早表达促进了先天表型的获得，这可以通过禁用 TCR 并依靠 CAR 来驱动分化来避免。延迟 CAR 表达并校准其在 TiPS 中的信号强度使得能够产生与 CD8αβ + 表现相似的人类 TCR ^{– CD8αβ} + ^CAR T 细胞来自外周血的 CAR T 细胞，在白血病小鼠模型中通过全身给药实现有效的肿瘤控制，并且不会引起移植物抗宿主病。在没有 TCR 的情况下，利用 CAR 驱动 TiPS 中的 T 细胞成熟可能有助于大规模开发有效的同种异体 CD8αβ ⁺ T 细胞，用于广泛的免疫疗法。