靶向CLL-1嵌合抗原受体T细胞的构建及其功能验证

OBJECTIVE: To explore the development of a CAR-T cells targeting CLL-1 and verify its function. METHODS: The expression levels of CLL-1 targets in cell lines and primary cells were detected by flow cytometry. A CLL-1 CAR vector was constructed, and the corresponding lentivirus was prepared. After infection and activation of T cells, CAR-T cells targeting CLL-1 were produced and their function was verified in vitro and in vivo. RESULTS: CLL-1 was expressed in acute myeloid leukemia（AML）cell lines and primary AML cells. The transduction rate of the prepared CAR T cells was 77.82％. In AML cell lines and AML primary cells, CLL-1-targeting CAR-T cells significantly and specifically killed CLL-1-expressing cells. Compared to untransduced T cells, CAR-T cells killed target cells and secreted inflammatory cytokines, such as interleukin-6 and interferon-γ, at significantly higher levels（P<0.001）. In an in vivo human xenograft mouse model of AML, CLL-1 CAR-T cells also exhibited potent antileukemic activity and induced prolonged mouse survival compared with untransduced T cells［not reached vs 22 days（95％ CI 19–24 days）, P＝0.002］. CONCLUSION: CAR-T cells targeting CLL-1 have been successfully produced and have excellent functions.