Fig. 2

Immunophenotyping analysis of monocytes/granulocytes in TdLNs and tumors from mice intraperitoneally administered anti-PD-1 antibody after inoculation with LLC1 cells or MC-38 cells. Anti-PD-1 mAb or isotype control (each 100 μg/body, n = 5) was administered a total of 3 times to mice every 3 days from 6 days after the inoculation with LLC1 cells and MC-38 cells (each 1 × 10⁶ cells/body). TdLNs and tumors were harvested 7 days after the final dosing, digested into single cell suspensions and analyzed by flow cytometry using fluorescence-conjugated antibodies. The average ratios of several monocytes/granulocytes, including DCs, MΦ and MDSCs, and non-monocytes/granulocytes in CD45⁺ cells from TdLNs (A) and tumors (B) are shown in pie charts with the respective ratios in parentheses. The percent differences between anti-PD-1 mAb treatment and isotype control-treatment for each monocyte/granulocyte in TdLNs and tumors were obtained by subtracting the average ratios of isotype control-treatment from those of anti-PD-1 mAb treatment in panel (C). The average ratios of CD80, CD86, MHC class I (MHC I), MHC class II (MHC II), CD197, and PD-L1 of several monocytes/granulocytes, including DCs, MΦ, and MDSCs, in CD45⁺ cells from TdLNs (D) and tumors (E) are shown in bar graphs. The average ratios of M1 phenotype MΦ (CD11c⁺CD206^–), M2 phenotype MΦ (CD11c^–CD206⁺), and MΦ in CD45⁺ cells from TdLNs (F) and tumors (G) are shown in bar graphs. Bar graphs depict the average values with SD (D–G). Significant differences are denoted with asterisks (*P < 0.05) (E, G).