Figure 2.

Administering the novel immune-enhancer intratumorally hinders B16F10 progression. (A) A cartoon illustrating the treatment regimen and the groups of mice treated with intratumoral injections. (B) Tumor weight (mg) measured on day 15 post B16F10 (2 mm³) implantation; each dot represents an individual tumor. Combined (C) and individual (D) tumor growth curves of B16F10 melanoma in the different mouse groups measured by caliper, as tumor volume mm³. (E) Density of CD8⁺ T cells in tumors as determined by flow cytometry. The densities were determined by dividing the total number of CD8⁺ T cells in each tumor by the tumor weight (in mg), pregated on TILs. Using FS/SS (F) FACS plots showing the total numbers of CD8⁺ T cells in a tumor, pregated on TILs. (G) Correlation between density of CD8⁺ T cells and tumor weight (mg). (H) Density of IFN-γ (I) and IFN-γ/TNF-α producing CD8⁺ T cells in tumors as determined by flow cytometry upon stimulation with 1 µg /mL PMA/ionomycin. The densities were determined by dividing the total number of CD8⁺ T cells in each tumor by the tumor weight (in mg). (J) Representative FACS plots showing the total numbers of IFN-γ/TNF-α producing CD8⁺ T cells in tumors. Statistical analysis (B, C, E, G–I) by ordinary one-way analysis of variance (multiple comparison) (mean±SEM). Statistical analysis in F by simple linear regression. n=12 (B–G) and n=5 (H, I). One representative of two similar experiments is shown. ****p<0.0001; ***p<0.001;, **p<0.01;, and *p<0.05. APC, antigen-presenting cell; IFN-γ, interferon gamma; MCT, microcrystalline tyrosine; ns, not significant; TIL, tumor-infiltrating lymphocyte; TNF-α, tumor necrosis factor alpha.