Fig. 5

HER2.28ζ/PD-L1.BB CAR-T cells have prolonged antitumor activity in vivo, with rapid and long-lasting eradication of pleural and peritoneal metastasis. a The schematic diagram of A549 and SKOV3^PD-L1 metastatic xenograft models in NSG mice inoculated via intrapleural or intraperitoneal (i.p.) injection, and treated 10 days later with CAR-T cells. b Representative bioluminescence (BLI) images of A549-Luc tumor growth in the pleural metastasis xenograft model. Mice were intrapleurally given 3 × 10⁶ CAR⁺ T or UTD cells, and tumor progression was monitored by in vivo imaging system (IVIS). c The BLI kinetics of A549-Luc tumor growth in the pleural metastasis xenograft model (n = 5). d Kaplan–Meier survival curve of mice after CAR-T cell treatment (n = 5). e Representative BLI images of SKOV3^PDL1-Luc tumor growth in the peritoneal metastasis xenograft model. Mice were intraperitoneally given 1 × 10⁶ CAR⁺ T or UTD cells, and tumor progression was monitored by IVIS. f The BLI kinetics of SKOV3^PDL1-Luc tumor growth in the peritoneal metastasis xenograft model (n = 5). g Kaplan–Meier survival curve of mice after CAR-T cell treatment (n = 5). h, i Summary of CAR-T cell (CD45⁺CD3⁺) number in the peritoneal cavity h and peripheral blood i of SKOV3^PD-L1-Luc peritoneal metastasis mice analyzed by flow cytometry 7 days after CAR-T cell treatment (n = 5). j The ratios of central memory T cells (CD45RO⁺CCR7⁺), PD-1⁺ k and PD-L1⁺ l CAR-T cells (n = 5). m The levels of effector cytokines (IFN-γ, IL-2 and TNF-α) in serum and peritoneal lavage fluid were measured by ELISA 7 days after CAR-T cells treatment (n = 5). n Quantification of CAR copy number by q-PCR to evaluate CAR-T cell persistence in the pleural cavity 2 weeks after SKOV3^PD-L1-Luc tumor regression (n = 3). P-values were determined by one-way ANOVA with Tukey’s multiple comparison test adjusted P value (c, f, h, i, j, k, l, m, n) or long-rank test d, g. *P < 0.05, **P < 0.01, ns, not significant. Data are expressed as the mean ± SD of each group