Fig 1: CD47 blockade therapy suppressed tumor progression in the PDX model.A–C A PDX tumor model was constructed using NSG mice using human HCC tissues. Schematic diagram showing the construction of a PDX tumor model and huPBMC injection through the tail vein. Mice (n = 6 for each group) were treated with anti-CD47 antagonist TTI-621 (8 mg/kg, three times a week for 3–4 weeks, i.p.) or vehicle (A). The tumor images (scale bar, 1 cm) (B), tumor weight (C) in the PDX tumor model using HCC tissues from patient 1. D–G The survival and tumor sizes of the tumor-bearing mice were recorded from patients 1–4. H, I mIF analysis of the tumor tissues in PDX model was used to detect the TIMs (CD11b), PD-L1, CD8+ T cells, and cytotoxic function (GZMB). Scale bar, 50 μm. J, K Western blot and qRT-PCR analysis were used to detect the PD-L1 expression in TIMs of tumor tissues in the PDX model. TIMs were obtained through magnetic bead sorting. All data presented are shown as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant.
Fig 2: SIRPα/CD47 axis promoted migration, PD-L1 expression of macrophages and HCC growth via upregulating PI3K/AKT signaling.A Schematic of the in vitro coculture experiments with PMA-treated THP-1 cells cocultured with Hepa1–6 or LPC-H12 cells to mimic liver cancer microenvironment treated with anti-CD47 mAb (10 μg/ml) or not. Macrophages were incubated with mCD47-Fc for 12 h before analysis. B Western blot analysis was used to detect the expression of indicated proteins in macrophages. C, D Transwell assay was used to detect the migration of macrophages towards the supernatants of the indicated groups. Scale bar, 100 μm. E Orthotopic HCC model using Hepa1–6 was established. Wild-type C57BL/6 mice (n = 6 for each group) were treated with anti-SIRPα mAb (100 μg/mouse, i.p.) and/or GSK690693 (30 mg/kg/day, i.p.) compared with the control group. Representative livers in each group were photographed at the endpoint (Scale bar, 1 cm). F, G Statistical analysis of tumor volume and tumor weight of the mice. All data presented are shown as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant.
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