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. 2022 Dec 15;10(12):e005002. doi: 10.1136/jitc-2022-005002

Figure 4.

Figure 4

Entinostat treatment increases the activation and cytotoxic capacity of antigen-specific T cells. (A) CD137 expression based activation of NBL-specific PRAMESLLQHLIGL-directed T cells after 48 hours of preincubation of GIMEN cells (left) or the patient-derived organoid 691b (right)±entinostat after overnight coculture at an E:T of 1:1. Entinostat concentration in GIMEN and no cytokine 691b condition was 5 µM; concentration in remaining 691b conditions was 2.5 µM. 100 U/mL IFN-γ or 1000 U/mL IFN-α was added to the indicated conditions. Data are shown from five replicates of three individual experiments. (B) IFN-γ (left) and TNF-α (right) secretions (pg/mL) in the supernatants of the overnight coculture between 691b organoids and PRAMESLLQHLIGL-directed T cells from (A) were determined by ELISA. Five replicates are shown from three individual experiments. (C) Luminescence-based cytotoxicity against luciferase-transduced 691b organoids after 48 hours of preincubation with 2.5 µM entinostat, 1000 U/mL IFN-α, or 100 U/mL IFN-γ. Cells were cocultured overnight at an E:T of 1:1. Data are shown from three individual experiments. Data are shown as mean±SEM. Statistical differences were calculated using the Mann-Whitney U test. **P<0.01, ****P<0.0001. E:T, effector-to-target ratio; IFN-α, interferon alpha; IFN-γ, interferon gamma; MFI, median fluorescent intensity; MHC-I, major histocompatibility complex class I; NBL, neuroblastoma; TNF-α, tumor necrosis factor alpha.