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Fig. 2 | Journal of Hematology & Oncology

Fig. 2

From: Targeting STAT3-VISTA axis to suppress tumor aggression and burden in acute myeloid leukemia

Fig. 2

Combination of VISTA mAb and STAT3 inhibitor displayed potent anti-leukemia effect. A In vivo bioluminescence imaging of xenograft mouse models (n = 6) with MOLM-13-EGFP/Luc cells treated with W1046 at dosage of 5 mg/kg and 15 mg/kg. B Survival curve of xenograft mouse models (n = 7) with MOLM-13-EGFP/Luc cells treated with W1046 at dosage of 5 mg/kg and 15 mg/kg. C The residual GFP + MOLM-13 cells in bone marrow of mice (n = 6) were detected by flow cytometry after being treated with W1046 for two weeks. D, E T cell-mediated cytotoxicity in co-culture system containing MOLM-13-EGFP/Luc cells and effector cells at indicated E/T ratio. The pretreated or unpretreated MOLM-13-EGFP/Luc cells were co-cultured with or without activated Jurkat cells stimulated by anti-CD3 antibody (1 μg/mL) and anti-CD28 antibody (3 μg/mL) at indicated E/T ratio with or without VISTA mAb (5 μg/mL) for 24 h, then the survival cells were measured by Steady-Glo. F CD8+ T cells population changed in the co-cultured system. The activated PBMCs stimulated by anti-CD3 antibody (1 μg/mL) and anti-CD28 antibody (3 μg/mL) were co-cultured with MOLM-13 cells (with or without W1046 pretreatment) at a E/T ration of 20:1 with or without VISTA mAb for 72 h, then the CD8+ T cells population were detected by flow cytometry. E/T ratio, E: Effector cells (Jurkat cells or PBMCs); T, Target cells (AML cells). G CFSE dilution assay to measure the proliferation of T cells. The pretreated or unpretreated MOLM-13 cells were co-cultured with activated Jurkat cells stimulated by anti-CD3 antibody (1 μg/mL) and anti-CD28 antibody (3 μg/mL) and stained by CFSE, then combining with or without VISTA mAb for 72 h. H Illustration of procedure of therapy in AML mouse models. C57BL/6 mice were inoculated IV with 3 × 106 C1498-EGFP/Luc cells and received indicated treatment. W1046 was intraperitoneally injected with 10 mg/kg every day and the VISTA mAb was intraperitoneally injected with 10 mg/kg once every other day. I In vivo bioluminescence imaging of xenograft mouse models (n = 6) with C1498-EGFP/Luc cells treated with W1046, VISTA mAb, or combination of W1046 and VISTA mAb. J Illustration of the procedure and identification of residual GFP + C1498 cells of mice (n = 6) by flow cytometry. K The residual GFP + C1498 cells in bone marrow of mice (n = 6) were detected by flow cytometry after being treated with W1046, VISTA mAb, or combination of W1046 and VISTA mAb for two weeks. L Survival curve of xenograft mouse models (n = 7) with C1498-EGFP/Luc cells treated with W1046, VISTA mAb, or combination of W1046 and VISTA mAb. M The infiltrating CD8+ T cells in bone marrow of mice (n = 6) were detected by flow cytometry after being treated with W1046, VISTA mAb, or combination of W1046 and VISTA mAb for two weeks. N Graphic Abstract (using the Biorender tools, an online platform for data analysis, https://biorender.com). Data were presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001

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