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. Author manuscript; available in PMC: 2021 Aug 1.
Published in final edited form as: Cancer Immunol Res. 2020 Dec 22;9(2):239–252. doi: 10.1158/2326-6066.CIR-20-0638

Figure 4. The pentatherapy regimen results in the activation of CEA-specific CD4+ and CD8+ T cells in the MC38-CEA tumor model.

Figure 4.

A. Splenocytes were collected on day 33 post-tumor inoculation and were analyzed via ELISA for IFNγ production after in vitro stimulation with an H2Kb-restricted CEA peptide. B. Proliferation of splenocytes from untreated, Ad-CEA+N-803, OX40+GITR, and the pentatherapy combination groups (n=5/group) was evaluated after ex vivo stimulation with I-Ab CEA peptide using a 3H-thymidine incorporation assay. C. Female C57BL/6-CEA-Tg mice (8–16 weeks old; n=10/group) were inoculated with 3×105 MC38-CEA cells on the flank. The tumor-bearing animals were treated with the pentatherapy regimen, N-803+OX40+GITR+IDOi (no Ad-CEA) or Ad-CEA+OX40+GITR+IDOi (no N-803). Tumor growth was monitored, and tumor volume and tumor growth control on day 28 are reported. D. For each treatment combination tested, the percentage of mice with tumor volume <300 mm3 was calculated and plotted against the number of IO agents received. Meta-analysis of four independent experiments is shown. E. Immune-related transcriptomes of tumor samples were analyzed using NanoString’s PanCancer Immune Profiling Panel. Heatmap showing select genes with data presented as fold-change values on scale of −20 (blue) to +3 (red). n.s., not significant. Error bars represent mean±SEM. These studies were repeated 3–4 times with similar results.