Extended Data Fig. 3: The CD3ζ-TIR-CAR-iMACs showed potent antitumor activity against intracranial GBM.
From: A second-generation M1-polarized CAR macrophage with antitumor efficacy
a, An overview of the anti-GBM assay of EGFRvIII-targeting CAR-iMACs in vivo. 1 × 104 FFluc+U87MGEGFRvIII cells were injected into the intracranial corpus striatum of NOD-SCID mice six days before immune cell therapy. On day -1, live animal imaging was conducted to monitor the formation of GBM. On day 0, EGFRvIII-targeting truncated CAR-iMACs and CD3ζ-TIR-CAR-iMACs with no IFN-γ/LPS-pretreatment were injected into the GBM lesion. On the following day 1, day 2, day 3, day 5, and day 10, the bioluminescent imaging was performed at the same time point to record the change of signal from the GBM. b, Visual presentation of the tumor signal from the intracranial GBM after treatment by the truncated CAR-iMACs and CD3ζ-TIR-CAR-iMACs. The PBS treatment group was designed as a negative control. c, Statistics analysis of tumor BLI signal from b (n = 5 mice from PBS group, n = 5 from truncated CAR-iMAC group, n = 5 mice from CD3ζ-TIR-CAR-iMAC group). The significance was calculated with the two-way ANOVA analysis and are presented as mean±s.e.m. p-value: ns, not significant, ***P < 0.001. d, The Kaplan-Meier survival curve of CAR-iMACs treated mice over 17 days from b. Significance was calculated with two-tailed log-rank Mantel-Cox test (CD3ζ-TIR-CAR-iMAC group versus PBS group, P = 0.0003. CD3ζ-TIR-CAR-iMAC group versus truncated CAR-iMAC group, P = 0.0003) All the statistic data above was shown by Graphpad Prism 8.2.1. e, The IHC staining showing the EGFRvIII protein expression of FFluc+U87MGEGFRvIII-derived GBM on day 8 and day 16 after the tumor cell inoculation, indicating the persistent expression of EGFRvIII antigen.