Figure 2.

CD39⁺ HBVs-CAR-T cells exerted potent antitumor activity in the HBV⁺ HCC organoid-T cell co-culture system

(A) The CD39⁺ and CD39⁻ HBVs-CAR T cells were sorted by FACS and co-cultured with autologous tumor organoids for 24 h separately. HBVs-CAR-T cells (red) were labeled with CellTrace Far Red and apoptotic cells (green) were labeled with caspase3/7 probe. Real-time Biotech imaging system was used to take 1 picture per hour for 24 h. GP120-CAR-T cells served as NC. Scale bars, 300 μm. (B) Summaries of quantitative statistics of HBVs-CAR-T cells (red) and apoptotic cells (green). Apoptotic cells (green) and infiltrating CAR-T cells (red) inside the area of organoids were calculated by the Spot function of Imaris software based on the size and intensity threshold. The initial number of infiltrating CAR-T cells at 0 h was defined as 0. ∗p < 0.05, ∗∗p < 0.005, ∗∗∗p < 0.0005 (1-way ANOVA). All data are means ± SEMs. Error bars represent SEMs of 3 view fields from each patient. (C) Representative flow cytometry analysis of organoid apoptosis by caspase3/7 probe (FITC).