Fig 6. GLA-SE improves IL12 induced tumor control, antigen-specific T cell expansion and myeloid cell activation.

(A, B) Mice bearing subcutaneous B16 melanoma tumors in their right flanks were treated initially with srRNA/mIL12 with or without the combination of the TLR4 agonist GLA-SE (n = 9–10). Mice that survived initial tumor challenge were rechallenged with a second identical tumor inoculation. Tumor growth was monitored with no further treatment (*p < 0.0001; representative data from 3 independent experiments). (C) Mice that survived until 66 days after rechallenge were sacrificed for splenocyte isolation. Splenocytes were stained for melanoma antigen-specific T cells populations (e.g. TRP1 and gp100) and analyzed using flow cytometry (p = 0.0456 and p = 0.005, respectively). (D) NanoString expression profiles of most upregulated genes in B16 tumors isolated from mice treated with srRNA/mIL12 with or without the combination of GLA-SE (n = 5). Tumor growth plots represent individual mice. One-way ANOVA analysis was used to determine significance of plots in C. Survival benefit was determined using Mantel-Cox Log-rank testing where p < 0.05 is considered significant. Gene analysis was conducted using the Advanced Analysis module of the nSolver software program.