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. Author manuscript; available in PMC: 2020 Aug 1.
Published in final edited form as: Clin Cancer Res. 2019 Oct 23;26(3):679–689. doi: 10.1158/1078-0432.CCR-19-2209

Figure 1 – Functional TIL targeting p15E failed to develop in the presence of OVA.

Figure 1 –

A, qPCR was used to measure p15E gene expression in MOC1ova and pMOC1 cells. p15E expression in MOC2 cells was used as a reference and set to 1.

B, TIL were cultured from day 14 MOC1ova or pMOC1 tumors. After 7 days in culture, TIL were assessed for tetramer positivity by flow cytometry. Representative dot plots are shown on the left. Quantity of tetramer positive TIL is shown on the right (n = 5 independent tumors each model).

C and D, TIL cultured from MOC1ova tumors (C) or pMOC1 tumors (D) were assessed for IFNγ production upon exposure to control (VSV-N52–59) or antigenic peptides (OVA257–264 and p15E604–611). Number of IFNγ producing cells was quantified by ELISpot (left, with representative photomicrographs of wells) and cumulative production of IFNγ was quantified by ELISA (right) (n = 5 independent tumors each model).

E, TIL cultured from day 14 MOC1ova or pMOC1 tumors were co-cultured with MOC1ova or pMOC1 target cells and loss of target cell viability was assessed in real-time by impedance analysis. Representative impedance plots are shown on the left. Quantification of loss of target cell index 12 hours after initiation of co-culture (vertical dashed line) is shown on the right (TIL pooled from 5 independent tumors each model).

F, MOC1ova tumors were established in WT B6 mice and completely resected at day 14. Forty days after resection, mice were challenged with either MOC1ova or pMOC1 cells and followed for tumor engraftment. Cumulative data from three independent experiments. Significance determined by Mantel-Cox analysis.

G, pMOC1 tumors were established in WT B6 mice and completely resected at day 14. Forty days after resection, mice were challenged with pMOC1 cells and followed for tumor engraftment. Cumulative data from two independent experiments.

Unless stated otherwise, all data shown is representative data from one of at least two independent experiments.

**, p < 0.01; ***, p < 0.001, student’s t-test; n/s, non-significant.