Figure 3. Effect of in vitro treatment of sorafenib on frequencies of T effector cells and Tregs.

Pretreatment samples of PBMCs from HCC patients were stimulated with anti-CD3/CD28 in the presence or absence of 10 μM concentration of sorafenib in vitro for 48 hours, and the frequencies of (A) CD4⁺CD127⁺PD-1⁺ T effector cells, (B) CD4⁺CD127⁺PD-1^– T cells, (C) CD4⁺Foxp3⁺ Tregs, (D) Foxp3⁺CTLA-4⁺ Tregs, (E) CD4⁺IFN-γ⁺ T cells, (F) CD8⁺IFN-γ⁺ T cells, (G) CD4⁺PD-1⁺VEGFR2⁺ T cells, and (H) CD8⁺PD-1⁺VEGFR2⁺ T cells were determined by flow cytometry as described in Methods. Each symbol represents an individual HCC patient: anti-CD3/CD28–treated (control) or sorafenib-treated PBMC. **P < 0.005, paired t test, †P < 0.003 statistically significant after Hommel multiplicity adjustment (n = 7 for A-F, n = 5 for G and H).