Extended Data Fig. 4: GPC3-targeting CAR-iMACs showed enhanced antitumor capacity against HCC.
From: A second-generation M1-polarized CAR macrophage with antitumor efficacy
a, The flow cytometry analysis was performed to determine the expression of GPC3-CARs on truncated CAR-iMACs, TIR-CAR-iMAC, and CD3ζ-TIR-CAR-iMACs by an APC-conjugated rabbit antibody specific to F(ab’)2 of human IgG. Shown is a representative result from three biological replicates. The results were processed using FlowJo. The numbers are mean±s.e.m. b, Real-time imaging by an Incucyte2022 A Live-Cell Analysis Instrument showed the signals of the tdTomato+ HepG2 cells (white arrow) during a time course of 138 hours after incubating GPC3-targeting truncated CAR-iMACs, TIR-CAR-iMAC, and CD3ζ-TIR-CAR-iMACs (bright field) with the tumor cells at the E/T ratio of 10/1. c, The TIR-based CAR-iMACs cooperating with CD47-antibody cured HepG2-derived hepatocellular neoplasm in tumor-bearing mice. 5 × 105 FFluc+HepG2 cells were intraperitoneally injected to B-NDG mouse. 4 days after that, GPC3-targeting truncated CAR-iMACs, TIR-CAR-iMAC, and CD3ζ-TIR-CAR-iMACs mixed with CD47-antibody were injected intraperitoneally into the tumor-bearing mice. 2 hours after the treatment, live animal imaging was performed to monitor the growth of HepG2 cells in vivo. The treatment group with only CD47-antibody was designed as a control. d, The statistics of change of tumor burden from the treated mice from c (n = 5 mice from CD47 antibody group, n = 5 mice from truncated CAR-iMAC + CD47 antibody group, n = 5 mice from TIR-CAR-iMAC + CD47 antibody group, n = 10 mice from CD3ζ-TIR-CAR-iMAC + CD47 antibody group). e, The statistics of change of weight of the mice from c. All the above data was shown by GraphPad Prism 8.2.1. The significance was calculated with a two-way ANOVA analysis with multiple comparisons and are presented as mean±s.e.m. p-value: ns, not significant, *P < 0.05,***P < 0.001,****P < 0.0001.