表达嵌合抗原受体（CAR）的T细胞是一种复杂且异质的基因治疗产品，其表型组成存在差异。通常，较高比例的未分化CAR T细胞与改善的抗肿瘤功能和存活能力相关。在本研究中，我们描述了一种新型的受体靶向的慢病毒载体（LV），名为62L-LV，该载体倾向于转导由L-选择素受体CD62L标记的未分化T细胞，并使CD4+和CD8+原代T细胞的转导率高达70％和50％。显著的是，相比VSV-LV，在长期培养中使用62L-LV转导并保持更多的未分化T细胞。有趣的是，脱落的CD62L既不会改变62L-LV颗粒与T细胞的结合，也不会影响其转导。经过仅24小时与62L-LV或VSV-LV激活的T淋巴细胞孵育2天就足以在白血病肿瘤小鼠模型中产生能够控制肿瘤生长的CAR T细胞。该数据证明通过短期离体暴露原代细胞于LV，可以产生具有强效的CAR T细胞。作为首个倾向于转导未分化T淋巴细胞的载体类型，62L-LV有潜力避免繁琐的T细胞亚型选择，并大大缩短CAR T细胞制造过程的时间。版权所有© 2023 Kapitza, Ho, Kerzel, Frank, Thalheimer, Jamali, Schaser, Buchholz和Hartmann。

Chimeric antigen receptor (CAR)-expressing T cells are a complex and heterogeneous gene therapy product with variable phenotype compositions. A higher proportion of less differentiated CAR T cells is usually associated with improved antitumoral function and persistence. We describe in this study a novel receptor-targeted lentiviral vector (LV) named 62L-LV that preferentially transduces less differentiated T cells marked by the L-selectin receptor CD62L, with transduction rates of up to 70% of CD4+ and 50% of CD8+ primary T cells. Remarkably, higher amounts of less differentiated T cells are transduced and preserved upon long-term cultivation using 62L-LV compared to VSV-LV. Interestingly, shed CD62L neither altered the binding of 62L-LV particles to T cells nor impacted their transduction. The incubation of 2 days of activated T lymphocytes with 62L-LV or VSV-LV for only 24 hours was sufficient to generate CAR T cells that controlled tumor growth in a leukemia tumor mouse model. The data proved that potent CAR T cells can be generated by short-term ex vivo exposure of primary cells to LVs. As a first vector type that preferentially transduces less differentiated T lymphocytes, 62L-LV has the potential to circumvent cumbersome selections of T cell subtypes and offers substantial shortening of the CAR T cell manufacturing process.Copyright © 2023 Kapitza, Ho, Kerzel, Frank, Thalheimer, Jamali, Schaser, Buchholz and Hartmann.