Figure 4.

IT heat-iOV-GM induces higher levels of IFN and proinflammatory cytokines and chemokines in distant non-injected tumors than live OV-GM. (A) Tumor implantation and treatment schedule in a bilateral intradermal tumor implantation model. (B) B16-F10 melanoma cells were implanted intradermally into the left and right flanks of C57BL/6J mice. After the tumors were established, the larger tumors on the right flank were injected with either PBS, live OV, live OV-GM, or heat-iOV-GM twice weekly. The non-injected tumors were harvested 2 days after the second injection and RNAs were extracted. Quantitative real-time PCR analyses of Ifnb, Il6, Ccl4, Ccl5, Cxcl9, and Cxcl10 gene expression in non-injected B16-F10 tumors isolated from mice treated with either PBS, OV, live OV-GM, or heat-iOV-GM (n=4-5, *P < 0.05, **P < 0.01, t-test). (C) Expression of IFN, proinflammatory cytokines, and chemokines in non-injected B16-F10 tumors from WT, Batf3^-/-, or STING^Gt/Gt mice treated with heat-iOV-GM were analyzed. Relative expression of Ifnb, Il6, Ccl4, Ccl5, Cxcl9, and Cxcl10 genes was measured by quantitative real-time RT-PCR and was normalized to the expression of GAPDH. Each panel shows the fold changes of the mRNA levels in non-injected tumors from WT, Batf3^-/-, or STING^Gt/GT mice treated with heat-iOV-GM, compared with those from WT mice treated with PBS (n=4, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). (D) Schematic diagram of a bilateral intradermal tumor implantation model with CD4 and/or CD8 depletion. (E) Relative expression levels of Ifnb, Il6, Ccl5, Cxcl10 in non-injected tumors from each treatment groups were measured by quantitative real-time RT-PCR and were normalized to the expression of GAPDH (n=4, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).