ABSTRACT
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 CD30-41BBζgroup , 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-γsecretion 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 .