Research on the effect of PD-L1 overexpression on CLL-1 CAR-T anti-acute myeloid leukemia
10.3760/cma.j.issn.0253-2727.2020.10.007
- VernacularTitle:PD-L1过表达对CLL-1 CAR-T细胞抗急性髓系白血病作用的影响
- Author:
Guoqiang LIN
1
;
Yanming ZHANG
;
Liqing KANG
;
Lei YU
;
Depei WU
Author Information
1. 苏州大学附属第一医院血液科，国家血液系统疾病临床医学研究中心，江苏省血液研究所，国家卫生健康委员会血栓与止血重点实验室 215006；徐州医科大学附属淮安医院，淮安市第二人民医院血液科 223002
- Keywords:
Leukemia, myeloid, acute;
PD-L1 overexpression;
C-type lectin-like molecule 1 (CLL-1);
Chimeric antigen receptor T cells
- From:
Chinese Journal of Hematology
2020;41(10):829-835
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of programmed death receptor ligand 1(PD-L1)on CLL-1 CAR-T against acute myeloid leukemia(AML).Methods:In this experiment, the PD-L1 expression vector was constructed, and then the lentivirus vector was packaged by three-plasmid packaging system. THP-1 monoclonal cell lines stably expressing PD-L1 were set up. The CLL-1 CAR-T was developed by our team, as the effector cell for co-culture with the THP-1 or THP1-PDL1 cell lines, respectively. Then, the LDH was tested using the kit, the supernatant cytokine was detected by CBA, and the CLL-1 CAR-T cell proliferation was demonstrated by flow cytometry(FCM)with CSFE labeled.Results:①The PD-L1 lentivirus vector was successfully constructed, and monoclonal cell lines of THP-1 with stable PD-L1 was set up and verified by FCM and PCR. ②The overexpression of PD-L1 inhibited CLL-1 CAR-T's ability to lyse THP-1 cells(E∶F ratio 10∶1); the killing efficiency of CLL-1 CAR-T on THP1-PDL1 cells was lower than that of THP-1 cells[(15.70±9.90)% vs(51.95 ± 2.52)%, P<0.05]. ③The overexpression of PD-L1 decrease the release of cytokine[THP1-PDL1 group vs THP-1 group: IFN-γ(115.66±3.13)pg/ml vs(1708.16 ± 26.76)pg/ml, P<0.05; IL-6(17.37±0.72)pg/ml vs(124.92±4.26)pg/ml, P<0.05; IL-10(5.69±0.13)pg/ml vs(124.12±3.02)pg/ml, P<0.05]. Additionally, the proliferation of CLL-1 CAR-T was also inhibited. Conclusion:Monoclonal cell lines of THP-1 with stable PD-L1 expression were successfully constructed, and the adverse effect of PD-L1 overexpression on CLL-1 CAR-T anti-AML was confirmed, which provided a theoretical basis for the regulation of CLL-1CAR-T through the PD-1/PD-L1 pathway.
