Figure 4 – Enhanced systemic anti-tumor immunity is observed after primary tumor resection.

A, illustration of the in vivo cytotoxicity experiment that explores selective killing of adoptively transferred cells pulsed with p15E, a MOC1 tumor antigen.

B, dot plots show the ratio of CFSE low to CFSE high cells and the percentage of p15E-specific killing in naïve, MOC1 tumor bearing or tumor resected (day 14 after surgery) recipient mice (n=5 mice for the naïve condition and n=7 mice total from two independent experiments for tumor bearing or tumor resected conditions). n/s, not significant; significance determined with ANOVA multiple comparisons test.

C, a connected line dot plot shows the growth of challenge tumors in tumor bearing or tumor resected mice (day 14 after surgery) with or without CD8 or CD4 cell depletion (or isotype control, n=10 mice total from two independent experiments per condition) beginning one day prior to tumor challenge (depletion continued twice weekly for 4 weeks). The vertical dotted line corresponds to day 35 after challenge tumor injection. Challenge tumors in tumor bearing mice were measured to day 35 only, since this is the point at which the primary tumors in these mice begin to reach end-point criteria, requiring euthanasia. Significance determined by 2-way ANOVA with Tukey’s multiple comparisons test.

D, a dot plot shows the tumor volumes 35 days after challenge tumor injection. Significance determined with ANOVA multiple comparisons test.

E, a Kaplan-Meier survival curve shows survival in tumor resected mice with or without CD8 or CD4 depletion. Significance was determined with a log-rank test.

F, pot plots show splenic Ly6G⁺ cells 24 hours after administration of a single dose of an isotype control antibody before (top) or the 1A8 antibody (bottom). Inset percentages show the CD11b⁺Ly6G⁺ fraction of all live splenic immune cells.

G, a connected line dot plot shows the growth of MOC1 primary tumors with or without Ly6G depletion (starting at day 19, continuing twice weekly for 2 weeks). Significance determined by 2-way ANOVA with Tukey’s multiple comparisons test.

H, a connected line dot plot shows the growth of MOC1 challenge tumors engrafted at day 20 into the contralateral flank of mice bearing primary MOC1 tumors treated with a Ly6G depletion or isotype control at day 10 (one day before challenge tumor engraftment). Y-axis shows days after challenge tumor implantation. Significance determined by 2-way ANOVA with Tukey’s multiple comparisons test.