Fig. 6. Lymphatic-sparing IO therapy mobilizes peripheral antitumor immunity.

a Normalized tumor volumes on day 9 or 10 after orthotopic transplantation of 10⁶ 4MOSC1 tumor cells and treatment in vivo with two doses of αCTLA-4 followed by either an early (red, n = 9) or late (blue, n = 10) neck dissection—day 6 versus day 11. b Principal component analysis plot from whole blood RNA sequencing, performed at day 10 after transplantation of 10⁶ 4MOSC1 tumor cells and treatment in vivo with two doses of αCTLA-4 followed by either an early (red) or late (blue) neck dissection—day 6 versus day 11, calculated and plotted with DESeq2, n = 5. c Left: Representative gene set enrichment mountain plots of differentially expressed genes; and right: corresponding heatmaps depicting the row normalized Z-score of the top differentially expressed genes from those gene sets identified in analysis of whole blood RNA sequencing after transplantation of 10⁶ 4MOSC1 tumor cells and treatment in vivo with two doses of αCTLA-4, followed by either an early (red), or late (blue) neck dissection. d Bubble plot illustrating the top hits from a Gene Ontology analysis, illustrating GO hits enriched in late vs early ND treatment sequencing groups as described in Fig. 5a (log₂FC > 1, adjusted P value < 0.05 upregulated genes identified in analysis of whole blood RNA sequencing; n = 5). e Cartoon describing the central role that the tumor-draining lymph node plays in the response to ICI therapy and the outcome of rational, lymphatic-preserving treatment sequencing in HNSCC; see text for details. The differences between experimental groups were analyzed using independent, two-sided t tests (a), DESeq2 (c) or Log₂FC P < 0.05 (d). All data represent averages ± SEM, except where indicated. ns   not statistically significant. Source data are provided as a Source Data file.