Investigating TOFU-DCs and Anti-PD-1 in GBM

Kluc or GL261 GBM tumor-bearing mice were treated starting on day 6 after the intracranial injection. The cohort of mice used in this experiment were mice that received no treatment (Untreated), mice which received control unloaded DCs in combination with anti-PD-1 Ab (Ctl-DCs + PD-1) or IgG Ab (Ctl-DCs + IgG), and mice which received TOFU mRNA-pulsed DCs in combination with anti-PD-1 Ab (TOFU-DCs + PD-1) or IgG Ab (TOFU-DCs + IgG) (n = 7 mice per group). Briefly, 5 × 10⁵ TOFU-DCs or Ctl DCs were injected intradermally along with anti-PD-1 or IgG Abs (10 mg/kg) (BioXcell, clones RMP1-14 and 2A3, respectively) delivered intraperitoneally. The DC vaccines were administered weekly for 3 weeks for a total of 3 vaccines. The anti-PD-1 and IgG Abs were administered to the mice every 3rd day for 2 weeks for a total of 5 doses. Bioluminescent imaging of the Kluc tumors was performed using luciferin substrate (Perkin Elmer LAS Inc) and the IVIS Spectrum Imaging System at the end of the treatment regimen on day 20 post the tumor implantation (n = 5 mice per group). For RNA-seq analysis of the tumor microenvironment, the animals were euthanized on day 21 after the tumor implantation (n = 4–5 mice per group) and the tumor tissue was processed for downstream analysis (see the “Tumor dissociation” section of the methods).