Cancer immunotherapy with autologous chimeric antigen receptor (CAR) T cells faces challenges in manufacturing and patient selection that could be avoided by using ‘off-the-shelf’ products, such as allogeneic CAR natural killer T (^AlloCAR-NKT) cells. Previously, we reported a system for differentiating human hematopoietic stem and progenitor cells into ^AlloCAR-NKT cells, but the use of three-dimensional culture and xenogeneic feeders precluded its clinical application. Here we describe a clinically guided method to differentiate and expand IL-15-enhanced ^AlloCAR-NKT cells with high yield and purity. We generated ^AlloCAR-NKT cells targeting seven cancers and, in a multiple myeloma model, demonstrated their antitumor efficacy, expansion and persistence. The cells also selectively depleted immunosuppressive cells in the tumor microenviroment and antagonized tumor immune evasion via triple targeting of CAR, TCR and NK receptors. They exhibited a stable hypoimmunogenic phenotype associated with epigenetic and signaling regulation and did not induce detectable graft versus host disease or cytokine release syndrome. These properties of ^AlloCAR-NKT cells support their potential for clinical translation.

使用自体嵌合抗原受体 (CAR) T 细胞进行癌症免疫治疗在制造和患者选择方面面临挑战，而使用“现成”产品可以避免这些挑战，例如同种异体 CAR 自然杀伤 T ( ^Allo CAR-NKT) 细胞。此前，我们报道了一种将人类造血干细胞和祖细胞分化为^Allo CAR-NKT细胞的系统，但由于三维培养和异种饲养细胞的使用阻碍了其临床应用。在这里，我们描述了一种临床指导方法，以高产量和纯度分化和扩增 IL-15 增强的^Allo CAR-NKT 细胞。我们生成了针对七种癌症的^Allo CAR-NKT 细胞，并在多发性骨髓瘤模型中证明了其抗肿瘤功效、扩展和持久性。这些细胞还选择性地耗尽肿瘤微环境中的免疫抑制细胞，并通过 CAR、TCR 和 NK 受体的三重靶向来拮抗肿瘤免疫逃避。它们表现出与表观遗传和信号调节相关的稳定的低免疫原性表型，并且不会诱导可检测到的移植物抗宿主疾病或细胞因子释放综合征。^Allo CAR-NKT 细胞的这些特性支持了其临床转化的潜力。