Fig 1: TICAs modulate the tumor immune microenvironment and drive T cell infiltration. (A) MC38 tumor bearing mice were treated with vehicle, 15 mg/kg EphA2/CD137 1:2 TICA (BCY12491), an enantiomeric non-binding control TICA (BCY13626) q3d intravenously or 2 mg/kg anti-CD137 agonist antibody (urelumab analog) q3d intraperitoneally individual tumor volumes normalized to tumor volume on the day of treatment initiation are shown grouped by treatment. *P<0.05, ***p<0.001 one-way ANOVA on day 6 comparing to vehicle. (B) Nanostring analysis of tumors show the immunomodulatory effect of BCY12491 and anti-CD137 agonist antibody on the 25 functional pathways including stimulation of NF-kB signaling, costimulatory signaling, cytokine/chemokine signaling, cytotoxicity and interferon signaling among others. Orange indicates high pathway scores; blue indicates low scores. Scores are displayed on the same scale via a Z-transformation. (C) Nanostring analysis of tumors show the effect of BCY12491 and anti-CD137 agonist antibody on the T cell (probe set: Cd3d, Cd3e, Cd3g, CD6, SH2D1A and Trat1), cytotoxic cell (probe set: Ctsw, Gzma, Gzmb, Klrb1, Klrd1, Klrk1, Nkg7 and Prf1) and macrophage (probe set: CD163, CD68, CD84 and Ms4a4a) content. (D) Representative images of tissue sections from tumors treated with vehicle, 15 mg/kg BCY12491, BCY13626 or 2 mg/kg anti-CD137 agonist antibody Q3D and stained for mouse CD8 are shown. (E) Nanostring analysis of tumors show the effect of BCY12491 and anti-CD137 agonist antibody on the checkpoint inhibitor Pdcd1 (protein PD-1), Cd274 (protein PD-L1) and CTLA4 (protein CTLA-4) transcription. (C, E) *P<0.05, ***p<0.001, one-way ANOVA with Dunnett’s post test. ANOVA, analysis of variance; PD-L1, programmed death ligand 1; TICAs, tumor-targeted immune cell agonists.