Journal: Frontiers in Immunology

Article Title: Targeting the LPS-STING axis: neomycin restores STING-mediated anti-tumor immune suppression and inhibits tumor growth

doi: 10.3389/fimmu.2025.1637667

Figure Lengend Snippet: Gram-negative bacteria drive tumor progression via LPS-induced immunosuppression. (A) Melanoma tumor-bearing mice (C57BL/6, n=4/group) received intratumoral injections of PBS (control), inactivated E coli (1×10 7 CFU), or LPS (1 mg/kg) every two days. Tumor volume was calculated by vernier caliper measurement (volume=length×width²×0.5), and data are presented as mean ± SEM. (B) Comparison of tumor weight at the end point. (C) Immunofluorescence staining of LPS was performed using specific markers to evaluate the abundance of LPS in tumor tissues among the Control group, E coli group and LPS group samples. LPS was labeled green and DAPI nuclear staining was blue. Data are shown as mean ± SD. (D) MC38 and B16-F10 cells were treated with different concentrations of LPS (0.1, 1, 10 μg/mL) for 6, 24, 48, and 72 hours. Cell viability (OD450 nm) was measured by CCK-8 assay at each time point. Data were normalized to 100% activity of the untreated group at the corresponding time point and expressed as mean ± SD (n = 3). Statistical analysis showed no significant cytotoxicity of LPS in either cell line across all tested conditions. *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: WT C57BL/6J mice were subcutaneously inoculated with B16-F10 melanoma cells (1.0×10 5 cells/mouse) to establish the tumor model. Three days post-inoculation, tumor-bearing mice were randomly assigned into four groups (n=4/group) (1): PBS control (2); Neomycin (30 mg/kg) (3); CMA (Cridanimod; 1.5 mg/kg, T5317, TargetMol); and (4) Neomycin + CMA combination.

Techniques: Bacteria, Control, Comparison, Immunofluorescence, Staining, Labeling, CCK-8 Assay, Activity Assay

Journal: Frontiers in Immunology

Article Title: Targeting the LPS-STING axis: neomycin restores STING-mediated anti-tumor immune suppression and inhibits tumor growth

doi: 10.3389/fimmu.2025.1637667

Figure Lengend Snippet: Neomycin inhibits tumor progression through targeting LPS and remodeling the immunosuppressive microenvironment. (A) Schematic representation of treatment model. The growth curve (B) and weight (C) of mice melanoma treated with 30 mg/kg neomycin, 1.5 mg/kg CMA, and their combination were compared with the control group. Comparisons were carried out at specific time intervals, the growth curve (D) and the weight (E) of colon carcinomas in mice treated with 30 mg/kg neomycin in contrast to the control group. (F) Immunofluorescence assays were performed to visualise LPS in both tumor and liver tissues. As shown in the figure, the left panel represents the control group, while the right panel corresponds to the treatment group. DAPI (shown in blue) was used as a nuclear counterstain. Magnification is indicated by a 50 μm scale bar. (G) The graph presents the quantified levels of LPS in mice serum samples. The x-axis indicates different experimental groups, which include the control group (B16) and the treatment group (B16+neomycin). (H) B16-F10 cells were treated with different concentrations of Neomycin (0.1, 0.5, 1, 5, 10 μg/mL) for 72 hours. Cell viability (OD450 nm) was measured by CCK-8 assay at each time point. (I) Representative immunofluorescence micrographs of MC38 tumor sections from control and neomycin-treated mice. Tissue sections were immunostained for the pan-macrophage marker F4/80 (red) and the M2-like polarization marker CD206 (green), with nuclear counterstaining by DAPI (blue). Scale bar: 50 μm. There are three replicates in each group, the error bars reflect the spread of three measurements. Data are shown as mean ± SD. Statistical significance was indicated by * with P < 0.05, ** with P < 0.01.

Article Snippet: WT C57BL/6J mice were subcutaneously inoculated with B16-F10 melanoma cells (1.0×10 5 cells/mouse) to establish the tumor model. Three days post-inoculation, tumor-bearing mice were randomly assigned into four groups (n=4/group) (1): PBS control (2); Neomycin (30 mg/kg) (3); CMA (Cridanimod; 1.5 mg/kg, T5317, TargetMol); and (4) Neomycin + CMA combination.

Techniques: Control, Immunofluorescence, CCK-8 Assay, Marker