Fig 1: Gut microbiome diversity is associated with response to PD-1 antibody immunotherapy in CT26 tumor-bearing mice. (A) Venn diagram of the total number of species shared between the three groups. (B) Hierarchical clustering tree on OUT level. (C) The community map of dominant species in three group at the level of species, the columns with different colors represent different species, and the length of the columns represents the proportion of species. (D) Bar plot of compositional differences at species level in the gut microbiome of three groups of mice by one-way ANOVA. (E) Circos of sample and species: a visual circle diagram describing the correspondence between samples and species.
Fig 2: Gut microbiota induces specific changes in plasma lipids and metabolome. (A) PCA graphs of the plasma metabolome from QC samples in positive and negative metabolomics analysis. (B) PCA graphs of the plasma lipidome from all samples. (C) PCA of plasma metabolites from after PD-1 antibody treatment group mice. (D) Heatmap shows the normalized relative abundances of metabolites with annotation which were significantly changed in Control vs. Coli group and Vanc vs. Coli group. (E) Correlations between the gut microbiota (at the species level) and potential plasma compounds. Cells are colored based on Pearson correlation coefficient between predominant bacteria (relative abundance) and metabolites (normalized abundance) in plasma. The red color represents a significant negative correlation (P < 0.05), the blue color represents a significant positive correlation (P < 0.05).
Fig 3: Compositional differences in the gut microbiome are relatively stable over time. (A) Venn diagram of the total number of species shared between the three subgroups in before PD-1 antibody treatment group. (B) Venn diagram of the total number of species shared between the three subgroups in after PD-1 antibody treatment group. (C) Unsupervised hierarchical clustering of top 50 species abundances in before PD-1 antibody treatment group. (D) Unsupervised hierarchical clustering of top 50 species abundances in after PD-1 antibody treatment group. (E) Bar plot of compositional differences at genus level in the gut microbiome of before PD-1 antibody treatment group mice by one-way ANOVA. (F) Bar plot of compositional differences at genus level in the gut microbiome of after PD-1 antibody treatment group mice by one-way ANOVA.
Fig 4: The expression of PD-1 in kidney tissue of mice. The IHC of PD-1 was shown in control groups (A), sham groups (B) and IRI groups (C). Quantification is represented in panel (D). Data are expressed as mean ± standard deviation. Difference between IRI group and sham group was made using unpaired Student’s t-test for normally distributed data. Images, × 200; original scale bar, 100 µm. **p<0.01. ***p<0.001.
Fig 5: Effect of PD-1 antibody immunotherapy on the diversity of CT26 tumor-bearing mice with different gut microbiome. (A) Schematic diagram of mouse model experiment process and sample collection and analysis. (B) Tumor size of mice at the end of immunotherapy. (C) Tumor size of mice at the end of immunotherapy in the presence of different antibiotic regimen in each group.
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