Fig. 2: Maintenance of response is not associated with specific immune profile.

a Flow cytometry analysis of T lymphocyte subpopulations in animals treated with IgG (black, n = 13 tumors) or anti-PD-1 (blue, n = 26 tumors). CD3⁺ cell count normalized to tumor weight (g), and relative abundance of CD4⁺ and CD8⁺ cells in the same tumors are shown (top). Abundance of effector/memory T cells (CD44^highCD62L^low) and naive T cells (CD44^lowCD62L^high) among CD4⁺ and CD8⁺ cells (bottom). Two-tailed Mann–Whitney U test was performed to determine statistical difference between the treatment groups. Circle, single tumor; horizontal line, mean; error bar, standard deviation; ns, not significant. b Representative immunofluorescence images and quantification of mouse tumors co-stained with DAPI (blue), CD3 (magenta), CD4 (green), CD8 (red), and PD-1 (yellow). Scale bar, 100 μm (top left picture) or 25 μm (top right picture). DR, green, n = 11 tumors; SR, purple, n = 4 tumors; NR, orange, n = 13 tumors. Two-tailed Mann–Whitney U test was performed to determine statistical difference between the response groups. Circle, single tumor; line, mean; ns, not significant. c Abundance of eight immune cell types from RNA-seq analysis in DR (green, n = 6 tumors), and NR (orange, n = 8 tumors) using MCP-counter algorithm²². Two-tailed Mann–Whitney U test was performed to determine statistical difference between the response groups. Triangle, single tumor; hinges, 25th and 75th percentiles; middle line, median; whiskers, minimum to maximum value.