Extended Data Fig. 2: CD3ζ-TIR-CAR-iMACs exhibited enhanced and persistent antitumor activity.
From: A second-generation M1-polarized CAR macrophage with antitumor efficacy
a, An overview of the antitumor assay of EGFRvIII-targeting CAR-iMACs in vivo. Four hours after injection of FFluc+U87MGEGFRvIII cells into NOD-SCID mice, EGFRvIII-targeting truncated CAR-iMACs, CD3ζ-CAR-iMACs, TIR-CAR-iMACs, and CD3ζ-TIR-CAR-iMACs without IFN-γ/LPS-polarization pretreatment were injected into the tumor-bearing mice. Two hours after the CAR-iMACs injection, animal imagings were conducted to detect the signal of BLI from tumor cells. On the following day 1, day 3, and day 7, bioluminescent imaging was performed at the same time during the days. b, Visual presentation of the tumor signal from the tumor-bearing mice after treatment by the four types of CAR-iMACs. c, Statistics analysis of tumor BLI signal from b (n = 5 mice from truncated CAR-iMAC group, n = 5 mice from CD3ζ-CAR-iMAC group, n = 10 mice from TIR-CAR-iMAC group, n = 10 mice from CD3ζ-TIR-CAR-iMAC group). Significance was calculated with a two-way ANOVA analysis and are presented as mean±s.e.m. p-value: ns, not significant, *P < 0.05, **P < 0.01. d, The Kaplan-Meier survival curve of CAR-iMACs treated mice over 29 days from b. Significance was calculated with two-tailed log-rank Mantel-Cox test (CD3ζ-CAR-iMAC group versus truncated CAR-iMAC group, P = 0.0999. TIR-CAR-iMAC group versus truncated CAR-iMAC group, P = 0.0003. CD3ζ-TIR-CAR-iMAC group versus truncated CAR-iMAC group, P = 0.0003) e, FCM analysis of CD80-positive population and CD163-positive population of EGFRvIII-targeting CD3ζ-CAR-iMACs, TIR-CAR-iMACs, and CD3ζ-TIR-CAR-iMACs isolated from the mice treated in the same way as in b for 48 hours. The FCM experiments were carried out in triplicates and the results were processed using FlowJo. f, Histograph analysis of e (n = 3 biologically independent samples per group). The significance was calculated with a two-way ANOVA analysis and are presented as mean±s.e.m. p-value: ns, not significant, *P < 0.05, **P < 0.01. All the statistical data above was shown by Graphpad Prism 8.2.1.
