Study on killing effect of HBsAg specific CAR-T cells on hepatocellular carcinoma / 中华肝胆外科杂志

ABSTRACT

Objective:To analyze the anti effect of chimeric antigen receptor (CAR)-T cells targeting hepatitis B surface antigen (HBsAg) on hepatocellular carcinoma cells.Methods:HBsAg-CAR gene was transduced into T cells (obtained from the blood of healthy donors) through a lentiviral vector. CD19-CAR-T cells were included as mock group, and untransduced T cells were included as control group. Cells of the three groups were co-cultured with hepatocellular carcinoma cells expressing HBsAg or not to detect the anti effect and releasing level of anti-tumor cytokines (tumor necrosis factor-α, interferon-γ, interleukin-2). Subcutaneous xenograft PLC/PRF/5 tumor model using NPG mice were established and HBsAg-CAR-T cells (experimental group, n=5) or untransfected T cells (control group, n=5) were injected through tail vein. Tumor volume was measured 15 days after injection. Results:HBsAg-CAR-T cells proliferation was good under in vitro culture, and the expression rate of CAR was stable. After co-cultured with hepatocellular carcinoma cells expressing HBsAg, the level of anti-tumor cytokines released by HBsAg-CAR-T cells was significantly higher than that of the other two groups of T cells, and the difference was statistically significant (all P<0.05); the anti rate of HBsAg-CAR-T cell group on HBsAg-positive hepatocellular carcinoma cells was significantly higher than that of the other two groups, and the difference was statistically significant (all P<0.05). The tumor volume of NPG mice in the experimental group was (250.8±62.8) mm 3, which was lower than that of the control group (757.5±102.6) mm 3, and the difference was statistically significant ( P<0.05). Conclusion:HBsAg-CAR-T cells can specifically recognize and kill HBsAg-positive hepatocellular carcinoma cells and release high level of anti-tumor cytokines.