Figure 3.
Tolinapant promotes immune-mediated tumor killing. (A) Secreted IL-2 level measured by Luminex assay after 5-day treatment of PBMC (3 donors) with 1 µM tolinapant in the presence or absence of 1 µg/mL SEB. Paired statistical analysis of the “dimethyl sulfoxide (DMSO) + SEB” vs “tolinapant + SEB” group, P = .0005 (paired t test). (B) As for panel A, except full-dose response to tolinapant in the presence or absence of SEB. (C) Unstimulated and stimulated CD8+ T-cell (from 5 donors) viability (viability dye eFluor 780) or proliferation (CellTrace Violet) after 5-day treatment with or without 10 µM tolinapant. (D) IL-2 or IL-10 level measured by Luminex assay after 5-day treatment of anti-CD3/CD28 stimulated CD8+ T cells (from 3 donors) with 1 µM tolinapant. *P < .05 (ratio paired t test). (E) A549-NucLight Red cell killing by anti CD3-activated PBMCs in the presence of tolinapant by IncuCyte. (F) Measurement of activated caspase-3 in A549-NucLight Red cells in F by IncuCyte. (G) A549-NucLight Red cell killing by anti-CD3/CD28-activated CD8+ T cells in the presence of tolinapant by IncuCyte. (H) Measurement of activated caspase-3 in A549-NucLight Red cells in H by IncuCyte. (I) Percentage cell viability of HH cells alone or HH cells cocultured with PBMCs treated with increasing concentrations of tolinapant without (left) or with (right) anti-CD3 stimulation after 48 hours (n = 3). (J) Secreted IL-2 and TNF-α levels measured by MSD assay from anti-CD3-stimulated HH-PBMC coculture assays treated with DMSO or 1 µM tolinapant for 72 hours. *P < .05 (ratio paired t test).

Tolinapant promotes immune-mediated tumor killing. (A) Secreted IL-2 level measured by Luminex assay after 5-day treatment of PBMC (3 donors) with 1 µM tolinapant in the presence or absence of 1 µg/mL SEB. Paired statistical analysis of the “dimethyl sulfoxide (DMSO) + SEB” vs “tolinapant + SEB” group, P = .0005 (paired t test). (B) As for panel A, except full-dose response to tolinapant in the presence or absence of SEB. (C) Unstimulated and stimulated CD8+ T-cell (from 5 donors) viability (viability dye eFluor 780) or proliferation (CellTrace Violet) after 5-day treatment with or without 10 µM tolinapant. (D) IL-2 or IL-10 level measured by Luminex assay after 5-day treatment of anti-CD3/CD28 stimulated CD8+ T cells (from 3 donors) with 1 µM tolinapant. *P < .05 (ratio paired t test). (E) A549-NucLight Red cell killing by anti CD3-activated PBMCs in the presence of tolinapant by IncuCyte. (F) Measurement of activated caspase-3 in A549-NucLight Red cells in F by IncuCyte. (G) A549-NucLight Red cell killing by anti-CD3/CD28-activated CD8+ T cells in the presence of tolinapant by IncuCyte. (H) Measurement of activated caspase-3 in A549-NucLight Red cells in H by IncuCyte. (I) Percentage cell viability of HH cells alone or HH cells cocultured with PBMCs treated with increasing concentrations of tolinapant without (left) or with (right) anti-CD3 stimulation after 48 hours (n = 3). (J) Secreted IL-2 and TNF-α levels measured by MSD assay from anti-CD3-stimulated HH-PBMC coculture assays treated with DMSO or 1 µM tolinapant for 72 hours. *P < .05 (ratio paired t test).

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