Fig. 4

Nb CAR-T cells specifically proliferate, activate and kill target cells.  A The expression of CD19 and CD20 on Raji, Daudi, and K562 cells. B The proliferation of Nb CAR-T cells. Cell Trace-labelled Nb CAR-T cells were incubated with mitomycin C-treated Daudi, Raji or K562 cells for 5 d, and then, the Cell Trace dilution was analysed by flow cytometry. CAR-T cells were gated on GFP signals. C The expression of CD69 on Nb CAR-T cells. Nb CAR-T cells were incubated with Daudi, Raji and K562 cells for 24 h, and then, the T cell activation marker CD69 was detected by flow cytometry. D, E The generation of Daudi-luc and K562-luc cell lines. Daudi and K562 cells were transduced with lentivirus encoding the luciferase gene, followed by puromycin selection. F, G Cytotoxicity of Nb CAR-T cells. Nb CAR-T cells were incubated with Daudi, K562 cells at an E:T of 2:1, and cell viability was determined via bioluminescence. H, I Cytokine production of Nb CAR-T cells. Nb CAR-T cells were incubated with Daudi or K562 cells overnight, and then, the culture supernatant was collected to measure IL-2. Data were analysed by one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant (p > 0.05)