The development of autologous chimeric antigen receptor T (CAR-T) cell therapies has revolutionized cancer treatment. Nevertheless, the delivery of CAR-T cell therapy faces challenges, including high costs, lengthy production times, and manufacturing failures. To overcome this, attempts have been made to develop allogeneic CAR-T cells using donor-derived conventional CD4 + or CD8 + T cells (T convs ), but severe graft-versus-host disease (GvHD) and host immune rejection have made this challenging. CD3 + CD4 − CD8 − double-negative T cells (DNTs) are a rare subset of mature T cells shown to fulfill the requirements of an off-the-shelf cellular therapy, including scalability, cryopreservability, donor-independent anticancer function, resistance to rejection, and no observed off-tumor toxicity including GvHD. To overcome the challenges faced with CAR-T convs , we evaluated the feasibility, safety, and efficacy of using healthy donor–derived allogeneic DNTs as a CAR-T cell therapy platform. We successfully transduced DNTs with a second-generation anti–CD19-CAR (CAR19) without hampering their endogenous characteristics or off-the-shelf properties. CAR19-DNTs induced antigen-specific cytotoxicity against B cell acute lymphoblastic leukemia (B-ALL). In addition, CAR19-DNTs showed effective infiltration and tumor control against lung cancer genetically modified to express CD19 in xenograft models. CAR19-DNT efficacy was comparable with that of CAR19-T convs . However, unlike CAR19-T convs , CAR19-DNTs did not cause alloreactivity or xenogeneic GvHD-related mortality in xenograft models. These studies demonstrate the potential of using allogeneic DNTs as a platform for CAR technology to provide a safe, effective, and patient-accessible CAR-T cell treatment option.

中文翻译：

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同种异体双阴性 CAR-T 细胞抑制肿瘤生长而无肿瘤外毒性

自体嵌合抗原受体 T (CAR-T) 细胞疗法的发展彻底改变了癌症治疗。然而，CAR-T细胞疗法的交付面临挑战，包括高成本、冗长的生产时间和制造失败。为了克服这个问题，已经尝试使用供体来源的常规 CD4 开发同种异体 CAR-T 细胞+或 CD8+T细胞（T转化率)，但严重的移植物抗宿主病 (GvHD) 和宿主免疫排斥反应使这变得具有挑战性。CD3+CD4-CD8-双阴性 T 细胞 (DNT) 是成熟 T 细胞的一个罕见子集，可满足现成细胞疗法的要求，包括可扩展性、冷冻保存性、独立于供体的抗癌功能、抗排斥性和未观察到脱落-包括GvHD在内的肿瘤毒性。克服CAR-T面临的挑战转化率，我们评估了使用健康供体来源的同种异体 DNT 作为 CAR-T 细胞治疗平台的可行性、安全性和有效性。我们成功地用第二代抗 CD19-CAR (CAR19) 转导了 DNT，而不妨碍它们的内源性特征或现成特性。CAR19-DNTs 诱导针对 B 细胞急性淋巴细胞白血病 (B-ALL) 的抗原特异性细胞毒性。此外，CAR19-DNTs 在异种移植模型中对转基因表达 CD19 的肺癌表现出有效的浸润和肿瘤控制。CAR19-DNT 疗效与 CAR19-T 相当转化率. 然而，不像CAR19-T转化率, CAR19-DNT 在异种移植模型中不会引起同种异体反应或异种 GvHD 相关的死亡率。这些研究证明了使用同种异体 DNT 作为 CAR 技术平台的潜力，可提供安全、有效且患者可使用的 CAR-T 细胞治疗选择。