Fig. 3: The IL-17 signaling-associated cellular microenvironment in melanomas treated with ICI.
a, Heatmap showing Spearman’s correlation between immune cell types (following the Bindea et al.51 algorithm) and IL-17A–IL-17F GES in tumor samples in bulk RNA-seq cohorts. b, Scatter dot plots showing estimated neutrophil cell enrichment in baseline tissue samples of therapy responders (n = 21) versus non-responders (n = 11) treated with dual ICI (left) and therapy responders (n = 19) versus non-responders (n = 22) treated with anti-PD-1 monotherapy (right) in the Gide et al.18 dataset. P values are from unpaired t-tests, and mean ± 95% CIs are plotted. Each dot represents a biologically independent sample. c, Kaplan–Meier plot for PFS according to baseline neutrophil cell enrichment levels (n = 10, ‘low’; n = 22, ‘high’, according to an optimal cut point) in the dual-ICI group of Gide et al.18. HRs and 95% CIs are reported for high-expression groups, and the P value is from the log-rank test. d,e, Schematic workflow (d) for LC-MS/MS analysis (e). LC/MS/MS icon created by BioRender.com. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis showing the top significantly enriched cellular pathways in TANs derived from CM tumors. The corresponding heatmap shows fold change in protein expression of IL-17 signaling pathway (according to KEGG mmu04657) components. f, Schematic workflow for naive BM neutrophil isolation and in vitro analysis. g, Quantitative PCR (qPCR) analysis of key IL-17 signaling components in untreated versus α-IL-17A-treated (5 µg ml−1) naive BM neutrophils cultured for 24 h in conditioned medium derived from CM mouse melanoma. Regular growth medium (no exposure to tumor cells) was used as a control. Bar plots represent mean + s.e.m. from n = 4 biologically independent samples; P values are from two-way ANOVA with Holm–Sidak’s multiple-comparison test. Shown is one representative of three independently performed experiments. h, Corresponding IL-17A (left, bar plot) and TH17 cytokine levels (right, heatmap) in the supernatants of BM neutrophils from g. The bar plot shows mean + s.e.m.; dots represent individual biological replicates. P values are from one-way ANOVA with Sidak’s multiple-comparison test. C.M., conditioned medium; R, responder; NR, non-responder; DC, dendritic cell; MACS, magnetic-activated cell sorting; max, maximum; min, minimum; TH, helper T cell; Treg, regulatory T cell.
