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The putative α7-selective nicotinic acetylcholine receptor (nAChR) agonists S24795 (S24) and PNU-282987 (PNU) inhibit the BzATP-mediated interleukin (IL)-1β release. Monocytic THP-1 cells and THP-1 cell-derived M1-like macrophages were primed for 5 h with lipopolysaccharide (LPS; 1 µg/ml). The P2X7 receptor agonist BzATP was added for another 40 min to trigger IL-1β release, which was quantified by ELISA (A,C) and by measuring IL-1α/β-equivalent <t>bioactive</t> concentrations using HEK-Blue™ IL-1R cells in a QUANTI-Blue™ assay (B, D) . The BzATP- (100 µM) induced release of IL-1β was investigated in the presence and absence of the nAChR agonists S24 (50 µM) and PNU (10 µM), acetylcholine (ACh, 10 µM) or phosphocholine (PC, 200 µM). The amount of IL-1β released in response to BzATP was calculated by subtracting the IL-1β concentrations of cells treated with LPS alone. In each experiment, the IL-1β concentrations obtained after stimulation with BzATP were set to 100% and all other values were calculated accordingly. Data are presented as individual data points, bars represent median, whiskers percentiles 25 and 75. *p ≤ 0.05, different from LPS-primed cells stimulated with BzATP alone. Friedman test followed by the Wilcoxon signed-rank test.
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The putative α7-selective nicotinic acetylcholine receptor (nAChR) agonists S24795 (S24) and PNU-282987 (PNU) inhibit the BzATP-mediated interleukin (IL)-1β release. Monocytic THP-1 cells and THP-1 cell-derived M1-like macrophages were primed for 5 h with lipopolysaccharide (LPS; 1 µg/ml). The P2X7 receptor agonist BzATP was added for another 40 min to trigger IL-1β release, which was quantified by ELISA (A,C) and by measuring IL-1α/β-equivalent bioactive concentrations using HEK-Blue™ IL-1R cells in a QUANTI-Blue™ assay (B, D) . The BzATP- (100 µM) induced release of IL-1β was investigated in the presence and absence of the nAChR agonists S24 (50 µM) and PNU (10 µM), acetylcholine (ACh, 10 µM) or phosphocholine (PC, 200 µM). The amount of IL-1β released in response to BzATP was calculated by subtracting the IL-1β concentrations of cells treated with LPS alone. In each experiment, the IL-1β concentrations obtained after stimulation with BzATP were set to 100% and all other values were calculated accordingly. Data are presented as individual data points, bars represent median, whiskers percentiles 25 and 75. *p ≤ 0.05, different from LPS-primed cells stimulated with BzATP alone. Friedman test followed by the Wilcoxon signed-rank test.

Journal: Frontiers in Immunology

Article Title: Putative α7-selective ligands interact with α9-containing nicotinic acetylcholine receptors and modulate immune functions of human mononuclear phagocytes

doi: 10.3389/fimmu.2026.1773637

Figure Lengend Snippet: The putative α7-selective nicotinic acetylcholine receptor (nAChR) agonists S24795 (S24) and PNU-282987 (PNU) inhibit the BzATP-mediated interleukin (IL)-1β release. Monocytic THP-1 cells and THP-1 cell-derived M1-like macrophages were primed for 5 h with lipopolysaccharide (LPS; 1 µg/ml). The P2X7 receptor agonist BzATP was added for another 40 min to trigger IL-1β release, which was quantified by ELISA (A,C) and by measuring IL-1α/β-equivalent bioactive concentrations using HEK-Blue™ IL-1R cells in a QUANTI-Blue™ assay (B, D) . The BzATP- (100 µM) induced release of IL-1β was investigated in the presence and absence of the nAChR agonists S24 (50 µM) and PNU (10 µM), acetylcholine (ACh, 10 µM) or phosphocholine (PC, 200 µM). The amount of IL-1β released in response to BzATP was calculated by subtracting the IL-1β concentrations of cells treated with LPS alone. In each experiment, the IL-1β concentrations obtained after stimulation with BzATP were set to 100% and all other values were calculated accordingly. Data are presented as individual data points, bars represent median, whiskers percentiles 25 and 75. *p ≤ 0.05, different from LPS-primed cells stimulated with BzATP alone. Friedman test followed by the Wilcoxon signed-rank test.

Article Snippet: IL-1α/β-equivalent bioactive concentrations (pg/ml) were determined by interpolation from a 4-parameter logistic standard curve generated with a serial dilution of recombinant human IL-1β (InvivoGen, Cat# rcyc-hil1b) that was dissolved in FBS-free RPMI 1640 medium and run in parallel with the samples.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay