Cytotoxic Effect And Optimization Of Different Antigen Epitope Of CD30-targeted Chimeric Antigen Receptor T Cells In Classical Hodgkin's Lymphoma Cell Line

Background: Classical Hodgkin's lymphoma (cHL) is one of the pathological types of Hodgkin's lymphoma, a kind of malignant tumor which originates in lymphatic hematopoietic system and mainly occurs in young and elder patients. So far, the effect of conventional chemotherapy is relatively good. However,patients suffering with refractor/relapsed cHL still need some other effective treatments. Therefore, the study on optimized and individualized treatments that can simultaneously reduce the complications becomes currently research focus. Chimeric antigen receptor-modified T cell (CART) is a rapid development of research hot spot in the field of immunotherapy in recent years. There have been certain achievements in the clinical treatment in our hospital about CD30-targeted CART immunotherapy, though the curative effect still need further optimized.Objective: In this study, we chose two kinds of monoclonal antibody (mAb) Ki-4 and Ber-H2 which have different antigen epitope to optimally construct different vectors carrying corresponding CAR gene fragment. Then to compare two kinds of CART cells'killing effects of cHL cell line L428 which expresses positive of CD30 and figure out whether its mechanism have some correlation with NF-?B signal path.Methods:?Construction of lentiviral vectors carrying CD30-CAR gene fragment with different antigen epitope.? Cultivation of CD30-targeted CART cell with different antigen epitope.?Comparison the two kinds of CART cells' killing effects of cHL cell line. ?Study on the impact of CD30-CARs on NF-?B mechanism of cHL cell line.Results: ?We have successfully constructed CD30-CART cells with different antigen epitope, about 65.7% shows positive. ?The mean killing effects of E:T=1:1 and 10:1 of Group Ki-4 was 35.15% and 66.99%; which in Group Ber-H2 was 31.95% and 56.11%;and in Group CD19 was 21.92% and 45.49%; while in Group CIK was 16.99% and 32.22%.P< 0.05,the difference has statistically significance. ?The result of Western Blotting shows that P-P65 of Group TNF- a?2h and 3h in Group Ki-4 were strongly positive, I ?B a of which were weakly positive; while P-P65 and I?B a of Group blank and Oh in Group Ki-4 as well as Group Ber-H2 were all weakly positive.Conclusions: ?CD30-CART cells with different antigen epitope have different killing effects on cHL cells. ?Ki-4-CARs were able to activate NF-?B signal path, but Ber-H2-CARs were not.