Figure 1. CD40ag combines with ICB to control growth of YR1.7 melanoma tumors.
(A) Schematic describing treatment regimen for survival experiments. WT C57Bl/6J mice were inoculated s.c. with YR1.7 melanoma cells at day 0 (2 tumors per mouse). Treatments by were administered by i.p. injection and were initiated on day 7 and repeated every three days, for a total of five doses. Tumor growth was monitored through 60 days post-tumor inoculation. Created with BioRender.com.
(B) Kaplan-Meier survival curves for YR1.7-bearing mice either left untreated (n=9, dark grey), or treated with high dose ICB (n=11, orange), low dose ICB (n=11, green), CD40ag monotherapy (n=13, red), or ICB lo + CD40ag (n=14, purple).
(C) Corresponding tumor volume (mm3) curves over time for survival data shown in (B).
(D) Tumor volume at day 10 (72 hours post-therapy initiation if applicable) for control, ICB lo-treated, ICB hi-treated, CD40ag-treated, and CD40ag+ICB lo-treated tumors. Summary data is represented as mean ± SD. *p < 0.05, ***p < 0.001 denotes statistical significance by one-way ANOVA with Holm-Sidak multiple comparisons correction (calculated in GraphPad).
(E) Two-dimensional UMAP embedding summarizing expression of an 11-dimensional panel of macrophage phenotype markers measured by flow cytometry by macrophages and monocytes (Live+CD45+CD3−NK1.1−B220−Ly6G−CD11b+CD24−) sorted from 8-day YR1.7 tumors either left untreated (black) or treated with CD40ag (red) (data collection at 24 hours post-treatment if applicable). Plots to the left are colored by treatment condition, while the plots to the right are colored by mean fluorescence intensity (MFI) for each indicated surface marker, showing in detail the rapid phenotypic changes induced by CD40ag in TAMs.