Fig. 5: Therapeutic effects of the engineered bacteria in the colonic orthotopic model.

A Scheme and grouping of in vivo therapy (n = 5). The colons of BALB/c mice were inoculated with CT-26-luc cells (1 × 10⁵ cells/mouse) on day -7, and the mice were treated with the indicated engineered bacteria (1 × 10⁸ CFU) by colon-specific administration on days 0 and 3, followed by AMF treatment (310 kHz and 23.8 kA/m) for 80 min at 24 h after colon administration. An anti-CD47 antibody (Anti-CD47ab; 20 mg/kg) and CD47nb (20 mg/kg) were i.p. injected on days 0 and 3 in G2 and G5, respectively. B Bioluminescence imaging to monitor tumor growth on days 0, 10, and 15. C Semi-quantitative results of the bioluminescence intensity of the tumor regions shown in panel B (n = 5 mice). The bioluminescence intensities of each mouse at days 10 and 15 were normalized to day 0. D The number of metastatic tumors in the abdomen were counted on day 15 (n = 5 mice). E, F The changes in red blood cell count (RBC; E) and hemoglobin (HGB; F) during therapy (n = 5 mice). G Flow cytometry analysis of Anti-CD47ab (blue) or CD47nb (red) on RBCs in BALB/c mice bearing CT-26-luc colonic orthotopic xenografts after a single treatment with Anti-CD47ab or Fe-Bac-HlpA/CD47nb-BLPs + AMF. The Anti-CD47ab and CD47nb were detected using a fluorescein-labeled antibody against IgG and a nanobody, respectively. RBC without staining was used as the blank, and RBCs from healthy mice without treatments was used as control (Con). H Quantitative results of mean fluorescence intensity (MFI) in panel G (n = 3 independent experiments). The data (C–F, H) are shown as the mean ± SD. Statistical analysis was performed by a two-tailed unpaired t test. ^*P < 0.05; ^**P < 0.01^{; ***}P < 0.001. Source data are provided as a Source Data file.