FIGURE 3 from Dual Blockade of EP2 and EP4 Signaling is Required for Optimal Immune Activation and Antitumor Activity Against Prostaglandin-Expressing Tumors
Year: 2023
Type: other
Abstract: TPST-1495 is more effective than a single EP4 or EP2 antagonists in overcoming PGE2-mediated immune suppression of recovery of cytokine production from human monocytes and T cells. TNFα ELISA from PGE2 blockade assay performed using human whole blood treated with TPST-1495 (A) or E7046 (B) in the presence of 10 nmol/L or 500 nmol/L PGE2. Graphs show one representative donor of 3 donors tested. C–E, IFNγ concentration in supernatants of PGE2 blockade assay performed on magnetically enriched T cells from human PBMC. Cells were treated with various concentrations of EP receptor inhibitors TPST-1495 (C), E7046 (D), or PF04418948 (E), followed by concentrations of PGE2 and CEF peptides as shown in C and D. Cytokine levels in supernatants were determined by bead-based cytokine array analysis. Graphs show data with N = 4 technical replicates from one of four independent experiments. Two different healthy donors were used across these experiments. F, Percent IFNγ production rescue, as normalized to the control (DMSO), from PGE2 Blockade assay supernatants. Magnetically enriched T Cells from human PBMC were treated with increasing amounts of EP1 (Ono-8130) and EP3 (L-798106) inhibitors and subjected to 333 nmol/L PGE2 before stimulation with CEF peptides. Each datapoint represents the average percent recovery from one experiment, with four total experiments shown. G, Percent IFNγ production rescue, as normalized to control with 0 nmol/L PGE2 and 0 nmol/L TPST-1495. Cells obtained as in C–F were treated with increasing TPST-1495 in the presence or absence of 10 mmol/L EP1 inhibitor and 10 mmol/L EP3 inhibitor. Each datapoint represents the average percent recovery from one experiment, with four total experiments shown. Mean and SD are depicted in all graphs and the Student two-sided t test was used to assess significance between individual groups. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05.
Language: English
Subfield: Pharmacology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4385658236