Fig. 5: Baseline immunological markers according to serum butyrate or propionate levels.

a, b Mean (SEM) percentage of blood Treg (FoxP3⁺ among CD4⁺ T cells; a) and mean (SEM) sCD25 (b) according to median concentration of serum butyrate (left panel) and propionate (right panel) concentrations before ipilimumab treatment (V1). c, d Pearson correlations between butyrate and propionate quantification in serum and percentages of peripheral Treg (c; n = 38) and sCD25 (d; n = 30) before ipilimumab treatment (V1). e Mean (SEM) percentage of Treg in human PBMC (n = 8) after 4 days of culture with or without anti-CD3 and escalating concentrations of butyrate. f Mean (SEM) concentrations of systemic TNFa (left panel) and MCP-1 (right panel) according to median concentration of serum butyrate. g Mean (SEM) of systemic TNFa (left panel) and MCP-1 (right panel) according to median concentration of serum propionate. h Mean (SEM) TNFa (left panel) and MCP-1 (right panel) secretions by human PBMC (n = 4) stimulated during 4 days with or without anti-CD3 and escalating concentrations of butyrate. i Spearman correlation between butyrate (left panel) and propionate (right panel) quantifications in the stool and zonulin in the serum from patients (n = 9, where both data were available) before ipilimumab treatment (V1). j Spearman correlation between zonulin and serum concentrations of TNFa (light gray squares) and MCP-1 (black squares) (n = 12, where both data were available). Each dot represents one patient; n (number of patients) are indicated on each graph otherwise stated in the legend. P-values are indicated on each graph; Two-tailed unpaired t test (a, b, f, g) and two-way RM ANOVA followed by Tukey’s tests for multiple comparisons was used (e and h). ns, not significant.