Objective:To construct a Luc+CD38-Raji cell line and perform the preliminary validation of its function to lay the foundation for the later exploration of the immune escape phenomenon at the CD38 locus in lymphoma cells.Methods:The CD38 gene locus of Luc+Raji cells was knocked down by CRISPR-cas9 technology and PiggyBac(PB)transposon system,and the Luc+CD38-Raji cell line was constructed,which was assayed by using flow cytometry to co-incubate with the Luc+CD38-Raji cell line at a 1:1 ratio of CD 19 CAR-T and CD38 CAR-T as well as the expression level of the surface activation factor CD69 on untransduced primary T cells,and luciferase assay was used to detect the killing efficiency of the above groups of effector cells against the Luc+CD38-Raji cell line.Re-sults:Luc+CD38-Raji cells were successfully constructed,and the results of activation experiments showed that both CD19 CAR-T and CD38 CAR-T could be activated by Luc+Raji cells.While Luc+CD38-Rajil9 monoclonal cells were only able to activate CD19 CAR-T due to the lack of CD38 expression.The results of killing experiments showed that both CAR-T cells were able to kill Luc+Raji cells,and the killing efficiency of CD38 CAR-T on Luc+CD38-Raji19 monoclonal cells was similar to that of the original T cell.Conclusions:The Luc+CD38-Raji cell line was successfully constructed,which lays the foundation for exploring the immune escape phenomenon at the CD38 locus of lymphoma at a later stage.
LymphomaCD38RajiCAR-TCRISPR-cas9PiggyBac(PB)transposon system
NETL
NSTL
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