Following the inoculation of KPC tumor cells by either orthotopic implantation or the hemispleen procedure, mice were treated with Cy (100 mg/kg) on Day 6, PEGPH20 (40μg/kg) on Day 6 and GVAX on Day 7. Flow cytometry was performed on tumor infiltrating lymphocytes (TIL) isolated from processed dissected orthotopic tumor or diffusely metastatic liver respectively. Each experimental group consisted of 3 or 4 mice analyzed individually. Relative percentage and total number of live CD3+CD8+ TIL in the combination PEPGH20/GVAX treatment group compared to single agent treatment groups in the (A) orthotopic model and (B) hemispleen model respectively. Relative percentage and total number of live CD3+CD4+ TIL in the combination PEGPH20/GVAX treatment group compared to single agent treatment groups in the (C) orthotopic model and (D) hemispleen model respectively. Data represent mean ± SEM from one representative experiment that was repeated three times. ns not significant, * p<0.05,** p<0.01, ***p<0.001. (E) Kaplan Meier survival curves of mice implanted with PDAC cells in untreated mice (n=22), mice treated with Cy/GVAX alone (n=18), PEGPH20 alone (n=18), or the combination PEGPH20/GVAX treatment (n=18). Data represent the combined data of two consecutive experiments. ns not significant, * p<0.05. (F) Kaplan Meier survival curves of mice implanted with PDAC cells in untreated mice (n=10) or mice treated with the combination PEGPH20/GVAX treatment with CD8+ T-cell depletion (n=10) and without CD8+ T-cell depletion (n=10). ns not significant,** p<0.01.