Figure 5

VTX-2337 and Cetuximab increase immune cell infiltration. Blood, tumors and spleens were harvested from a subset of mice (n = 4–5) one day after drug treatment and analyzed for serum concentrations of TNFα (A), and tumor (B) and splenic (C) lymphoid DCs, CD69 + NK cells (D) and IFNγ + CD4 + T cells (E) using flow cytometry. (F) Images represent CD8 immunostaining in control (left image) and VTX + CTX (right image)-treated tumors. Bars = 100 µm (inset bars = 20 µm). (G–J) Female BALB/c mice (n = 9–10 mice/group) bearing TUBO-hEGFR tumors were treated with VTX in combination with CTX with or without anti-CD4, anti-CD8 or anti-asialo-GM1 (anti-NK) and tumor growth (G) was measured. Spleens were analyzed by flow cytometry for validation of CD4⁺ T cell (H), CD8⁺ T cell (I) and NK cell (J) depletion. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns: not significant (One-way ANOVA, Tukey’s multiple comparison test). Error bars represent standard deviation from the mean.