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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 7. Ad-CEA, N-803, OX40, GITR, and IDOi combination promotes effector CD4+ T cell populations and inhibits Treg suppression.

Figure 7.

A. Spleens and tumors from Fig. 6A were evaluated for CD4+ T cells and CD4+FoxP3+ Tregs via flow cytometry (gated on live, CD3+). B. Tumor sections were evaluated for CD4 and FoxP3-expressing cells via immunofluorescence. C-D. CD4+ T cells were further analyzed by flow cytometry to determine (C) Ki67+ and (D) CD44+ and CD44+CD62L+ populations. E. Tregs isolated from untreated and pentatherapy-treated 4T1 primary tumors harvested 28 days post-tumor implantation were cocultured with CD4 cells purified from spleens of naïve mice. CD4+ T-cell proliferation was determined via 3H-thymidine incorporation assay. F. Epacadostat, tryptophan, and kynurenine in the serum of the untreated and pentatherapy-treated MC38-CEA tumor-bearing animals were determined via HPLC. One-way ANOVA or Student t-test. *P<0.05; **P<0.01; ***P<0.005, ****P<0.001. Error bars represent mean±SEM. These studies were repeated 3–4 times with similar results.