Several obstacles to the production, expansion and genetic modification of immunotherapeutic T cells in vitro have restricted the widespread use of T-cell immunotherapy. In the context of HSCT, delayed naïve T-cell recovery contributes to poor outcomes. A novel approach to overcome the major limitations of both T-cell immunotherapy and HSCT would be to transplant human T-lymphoid progenitors (HTLPs), allowing reconstitution of a fully functional naïve T-cell pool in the patient thymus. However, it is challenging to produce HTLPs in the high numbers required to meet clinical needs. Here, we found that adding tumor necrosis factor alpha (TNFα) to a DL-4-based culture system led to the generation of a large number of nonmodified or genetically modified HTLPs possessing highly efficient in vitro and in vivo T-cell potential from either CB HSPCs or mPB HSPCs through accelerated T-cell differentiation and enhanced HTLP cell cycling and survival. This study provides a clinically suitable cell culture platform to generate high numbers of clinically potent nonmodified or genetically modified HTLPs for accelerating immune recovery after HSCT and for T-cell-based immunotherapy (including CAR T-cell therapy).

体外免疫治疗性 T 细胞的生产、扩增和遗传修饰的几个障碍限制了 T 细胞免疫疗法的广泛使用。在 HSCT 的背景下，幼稚 T 细胞恢复延迟会导致不良结果。一种克服 T 细胞免疫疗法和 HSCT 主要局限性的新方法是移植人类 T 淋巴祖细胞 (HTLP)，从而在患者胸腺中重建功能齐全的幼稚 T 细胞库。然而，生产满足临床需求所需的大量 HTLP 具有挑战性。这里，我们发现将肿瘤坏死因子 α (TNFα) 添加到基于 DL-4 的培养系统中会导致产生大量未修饰或基因修饰的 HTLP，这些 HTLP 具有来自 CB HSPC 的高效体外和体内 T 细胞潜能或 mPB HSPCs 通过加速 T 细胞分化和增强 HTLP 细胞循环和存活。本研究提供了一个临床适用的细胞培养平台，以生成大量临床有效的非修饰或基因修饰的 HTLP，以加速 HSCT 后的免疫恢复和基于 T 细胞的免疫疗法（包括 CAR T 细胞疗法）。