Objective To develop a CD30-targeted CAR-T cell drug based on the multi-chain chimeric antigen re-ceptor T cells(CAR-T)of the bridging protein DAP12,and to study the in vitro and in vivo preclinical efficacy of CD30 CAR-T on Hodgkin lymphoma tumor cells.Methods Through gene synthesis and molecular cloning tech-niques,a CAR plasmid targeting CD30 was designed and constructed,and the obtained lentivirus was packaged.The T cells were transfected with the lentivirus,where the multi-chain CAR-T targeting CD30 was the CD30-KIRS2/Dap12-BB group,the single-chain second-generation CAR-T was the CD30-41BBζ group,and the T cells without virus infection were the NTD group.The positive rate of CAR was detected by flow cytometry,the cytotoxic-ity of the cells was detected by lactate dehydrogenase(LDH)release assay,the secretion level of the cytokine in-terferon γ(IFN-γ)was detected by enzyme-linked immunosorbent assay(ELISA),and the antitumor activity of CD30 CAR-T in mice was further detected by a mouse xenograft tumor model.Results A comparison was made between the multi-chain CAR-T targeting CD30 and the single-chain second-generation CAR-T.It was found that the antitumor effect of the multi-chain CAR-T was similar to that of the single-chain CAR-T.However,it was worth noting that the IFN-γ secretion level of the multi-chain CAR-T was higher(P<0.001).More importantly,in the mouse tumor model experiment,the multi-chain CAR-T achieved complete tumor regression.Conclusion The multi-chain CAR-T targeting CD30 is superior to the traditional single-chain CAR-T in terms of antitumor activity.
关键词
嵌合抗原受体修饰的T细胞/CD30/霍奇金淋巴瘤/DAP12/过继性细胞疗法
Key words
chimeric antigen receptor modified T cells/CD30/Hodgkin lymphoma/DAP12/adoptive cell thera-py
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