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. 2021 Apr 8;6(7):e141912. doi: 10.1172/jci.insight.141912

Figure 5. HPV-specific T cells in the TME and periphery induced by PRGN-2009.

Figure 5

C57BL/6 mice (n = 6–7 per group) bearing s.c. TC-1 HPV16+ tumors were treated with empty vector control (1 × 109 VP, s.c.) or PRGN-2009 (1 × 109 VP, s.c.) on days 7 and 14 after tumor implantation. At the end of study (day 23), tumors were harvested. CD45+ TILs were isolated and stimulated overnight in triplicate wells with combined overlapping HPV16 E6/E7 15-mer peptides or DMSO (negative control), and they were then evaluated by flow cytometry. (A–F) Graphs show the number of cells/mg tumor for total CD8 (A), IFN-γ–producing CD8 (B), IFN-γ plus GzmB–producing CD8 (C), total CD4 (D), IFN-γ–producing CD4 (E), and IFN-γ plus GzmB–producing CD4 (F). (G) Unstimulated TILs were also evaluated for CD8 and HPV16 E7 tetramer. (H and I) An ELIspot assay was performed on splenocytes using overlapping 15-mer peptides for HPV16 E6 (H) and HPV18 E6 (I). Medians are shown. Mann-Whitney U test was used. *P < 0.05, **P < 0.01. TILs, tumor-infiltrating lymphocytes; TME, tumor microenvironment; VP, virus particles.