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. Author manuscript; available in PMC: 2017 Jan 15.
Published in final edited form as: Clin Cancer Res. 2015 Aug 31;22(2):436–447. doi: 10.1158/1078-0432.CCR-15-1070

Figure 6. Analysis of TIL from the anti-PD1 animal study.

Figure 6

At Day 42 in the study shown in Figure 5, flank tumors from Ly95 and Ly95 + PD1 Ab treated mice were harvested, digested and processed into single-cell suspension.

A) FACS analysis using anti-hCD45 (upper panels) revealed equal degree of TIL infiltration. FACs analysis using the TCRVβ13.1 antibody, showed an equal percentage of Ly95 TCR+ cells in both groups. Dot-plots display representative analyses from experiment with pooled samples. Bar graph displays averaged results ± S.E. from independently experiment repeats analyzing pooled samples.

B) Purified TILs (see Figure 2 Legend) from each group were cocultured with A549-A2-ESO tumors at 20:1 E:T ratio for 18hrs. TILs from the Ly95 group demonstrated profound hypofunctional killing and cytokine secretion ability. TILs from the Ly95 + anti-PD1 antibody group demonstrated significantly better killing and cytokine secretion ability. When 10ug/ml of anti-human PD1 Ab was added to the coculture of Ly95 TIL, killing ability was partially restored (p<0.05). However, TIL function even when exposed to PD1 Ab was not has high as that of TILs isolated from Ly95 + PD1 Ab treated group (* p<0.05, ** p<0.01, *** p<0.001). Bar graphs display average values ± S.E. from triplicates.