Fig. 3

Circulating S100A9⁺CD14⁺ monocyte predicts unfavorable responses to anti-PD-1 immunotherapy. a UMAP visualization of immune cells between NR and R. S100A9⁺CD14⁺ monocytes are boxed with red dashed lines. b Percentage of each immune subset between the NR group (blue) and R group (yellow), represented as box-whisker plots. Significance was evaluated using 2-way ANOVA. c Violin plots representing the percentage of S100A9⁺CD14⁺ monocytes in each patient. d Survival curves showing overall survival stratified by the percentage of S100A9⁺CD14⁺ monocyte. e Left: Representative image of dual immunofluorescence demonstrating S100A9 (red) and CD14 (green) positive cells and S100A9⁺CD14⁺ monocytes (white triangles) in HCC tumors of a responder and a non-responder. Right: Number of S100A9⁺CD14⁺ monocytes quantified in five randomly selected fields per patient (n = 4 in NR; n = 4 in R). Scale bar, 50 μm. f Levels of plasma S100A9 in patients among different ICB best efficacy. g ROC curves and the area under the curve (AUC) of the plasma S100A9 level (red) or NLR (dashed blue) for discrimination between responders and non-responders. The cut-off value for S100A9 level was 308.110 ng/ml. h Best efficacy of patients stratified by plasma S100A9 levels. i Survival curves showing progress-free survival stratified by the plasma S100A9 level. P value in (c and e) were calculated using unpaired Student’s t-test. P values in (d and i) were calculated using the log-rank test