In Vivo Generation of CAR T Cells Selectively in Human CD4+ Lymphocytes
In Vivo Generation of CAR T Cells Selectively in Human CD4+ Lymphocytes
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DOI：
10.1016/j.ymthe.2020.05.005
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发表时间：
2020-08-05
影响因子：
12.4
通讯作者：
Buchholz, Christian J.
中科院分区：
文献类型：
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作者：
Agarwal, Shiwani;Hanauer, Julia D. S.;Buchholz, Christian J.
CD19特异性嵌合抗原受体(CARS)修饰的T细胞为白血病患者带来了显著的临床益处，但对制造构成了挑战。我们最近已经证明了使用CD8靶向慢病毒载体(CD8-LV)在体内产生CD19-CAR T细胞。在这项研究中，我们研究了使用CD4靶向LV(CD4-LV)在体内产生CD4(+)CART细胞。将人外周血单个核细胞(PBMC)移植到NSG小鼠体内，发现40%-60%的人CD4(+)淋巴细胞呈CAR阳性，而CD8(+)细胞仍为CAR阴性。CAR(+)T细胞表现为辅助性T细胞1(Th1)/Th2表型，并伴有CD19(+)B细胞清除。用人CD34(+)细胞构建的NSG小鼠静脉注射CD4-LV后，在2-3周内可诱导CAR表达和B细胞清除。在肿瘤小鼠模型中的临床前分析显示，与单独注射CD8-LV或作为CD4-LV的混合物注射的小鼠相比，注射CD8-LV的小鼠表现出更快和更好的肿瘤细胞杀伤能力。进一步分析表明，CD4(+)CAR(+)细胞可能优于CD8(+)CAR(+)细胞，尤其是在靶抗原负荷较高的情况下，这主要是因为CD8细胞更容易耗尽。这第一次描述了体内产生的CD4(+)CAR T细胞，支持了它们在细胞治疗中的重要性。
T cells modified with CD19-specific chimeric antigen receptors (CARs) result in significant clinical benefit for leukemia patients but constitute a challenge for manufacturing. We have recently demonstrated the in vivo generation of CD19-CAR T cells using the CD8-targeted lentiviral vector (CD8-LV). In this study, we investigated the in vivo generation of CD4(+) CART cells using CD4-targeted LV (CD4-LV). Administration of CD4-LV into NSG mice transplanted with human peripheral blood mononuclear cells (PBMCs) led to 40%-60% of human CD4(+) lymphocytes being CAR positive while CD8(+) cells remained CAR negative. CAR(+) T cells displayed a T helper 1 (Th1)/Th2 phenotype, which was accompanied by CD19(+) B cell elimination. Intravenous administration of CD4-LV into NSG mice reconstituted with human CD34(+) cells induced CAR expression and B cell elimination within 2-3 weeks post-injection. Preclinical analysis in a tumor mouse model revealed that mice administered CD4-LV exhibited faster and superior tumor cell killing compared to mice injected with CD8-LV alone or as a mixture with CD4-LV. Further analysis suggests that CD4(+)CAR(+) cells may outperform CD8(+)CAR(+) cells, especially at a high burden of target antigen, mainly since CD8 cells are more prone to exhaustion. This first description of in vivo-generated CD4(+) CAR T cells supports their importance for cellular therapy.