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Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes
OBJECTIVE: The pro-inflammatory cytokine interleukin-1β (IL-1β) plays a central role in host defense against infections. High systemic IL-1β levels, however, promote the pathogenesis of inflammatory disorders. Therefore, mechanisms controlling IL-1β release are of substantial clinical interest. Rece...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10034071/ https://www.ncbi.nlm.nih.gov/pubmed/36969210 http://dx.doi.org/10.3389/fimmu.2023.1140592 |
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author | Richter, Katrin Asci, Nilay Singh, Vijay K. Yakoob, Sanaria Hawro Meixner, Marion Zakrzewicz, Anna Liese, Juliane Hecker, Andreas Wilker, Sigrid Stumpf, Sabine Schlüter, Klaus-Dieter Rohde, Marius Gödecke, Axel Padberg, Winfried Manzini, Ivan Schmalzing, Günther Grau, Veronika |
author_facet | Richter, Katrin Asci, Nilay Singh, Vijay K. Yakoob, Sanaria Hawro Meixner, Marion Zakrzewicz, Anna Liese, Juliane Hecker, Andreas Wilker, Sigrid Stumpf, Sabine Schlüter, Klaus-Dieter Rohde, Marius Gödecke, Axel Padberg, Winfried Manzini, Ivan Schmalzing, Günther Grau, Veronika |
author_sort | Richter, Katrin |
collection | PubMed |
description | OBJECTIVE: The pro-inflammatory cytokine interleukin-1β (IL-1β) plays a central role in host defense against infections. High systemic IL-1β levels, however, promote the pathogenesis of inflammatory disorders. Therefore, mechanisms controlling IL-1β release are of substantial clinical interest. Recently, we identified a cholinergic mechanism inhibiting the ATP-mediated IL-1β release by human monocytes via nicotinic acetylcholine receptor (nAChR) subunits α7, α9 and/or α10. We also discovered novel nAChR agonists that trigger this inhibitory function in monocytic cells without eliciting ionotropic functions at conventional nAChRs. Here, we investigate the ion flux-independent signaling pathway that links nAChR activation to the inhibition of the ATP-sensitive P2X7 receptor (P2X7R). METHODS: Different human and murine mononuclear phagocytes were primed with lipopolysaccharide and stimulated with the P2X7R agonist BzATP in the presence or absence of nAChR agonists, endothelial NO synthase (eNOS) inhibitors, and NO donors. IL-1β was measured in cell culture supernatants. Patch-clamp and intracellular Ca(2+) imaging experiments were performed on HEK cells overexpressing human P2X7R or P2X7R with point mutations at cysteine residues in the cytoplasmic C-terminal domain. RESULTS: The inhibitory effect of nAChR agonists on the BzATP-induced IL-1β release was reversed in the presence of eNOS inhibitors (L-NIO, L-NAME) as well as in U937 cells after silencing of eNOS expression. In peripheral blood mononuclear leukocytes from eNOS gene-deficient mice, the inhibitory effect of nAChR agonists was absent, suggesting that nAChRs signal via eNOS to inhibit the BzATP-induced IL-1β release. Moreover, NO donors (SNAP, S-nitroso-N-acetyl-DL-penicillamine; SIN-1) inhibited the BzATP-induced IL-1β release by mononuclear phagocytes. The BzATP-induced ionotropic activity of the P2X7R was abolished in the presence of SIN-1 in both, Xenopus laevis oocytes and HEK cells over-expressing the human P2X7R. This inhibitory effect of SIN-1 was absent in HEK cells expressing P2X7R, in which C377 was mutated to alanine, indicating the importance of C377 for the regulation of the P2X7R function by protein modification. CONCLUSION: We provide first evidence that ion flux-independent, metabotropic signaling of monocytic nAChRs involves eNOS activation and P2X7R modification, resulting in an inhibition of ATP signaling and ATP-mediated IL-1β release. This signaling pathway might be an interesting target for the treatment of inflammatory disorders. |
format | Online Article Text |
id | pubmed-10034071 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-100340712023-03-24 Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes Richter, Katrin Asci, Nilay Singh, Vijay K. Yakoob, Sanaria Hawro Meixner, Marion Zakrzewicz, Anna Liese, Juliane Hecker, Andreas Wilker, Sigrid Stumpf, Sabine Schlüter, Klaus-Dieter Rohde, Marius Gödecke, Axel Padberg, Winfried Manzini, Ivan Schmalzing, Günther Grau, Veronika Front Immunol Immunology OBJECTIVE: The pro-inflammatory cytokine interleukin-1β (IL-1β) plays a central role in host defense against infections. High systemic IL-1β levels, however, promote the pathogenesis of inflammatory disorders. Therefore, mechanisms controlling IL-1β release are of substantial clinical interest. Recently, we identified a cholinergic mechanism inhibiting the ATP-mediated IL-1β release by human monocytes via nicotinic acetylcholine receptor (nAChR) subunits α7, α9 and/or α10. We also discovered novel nAChR agonists that trigger this inhibitory function in monocytic cells without eliciting ionotropic functions at conventional nAChRs. Here, we investigate the ion flux-independent signaling pathway that links nAChR activation to the inhibition of the ATP-sensitive P2X7 receptor (P2X7R). METHODS: Different human and murine mononuclear phagocytes were primed with lipopolysaccharide and stimulated with the P2X7R agonist BzATP in the presence or absence of nAChR agonists, endothelial NO synthase (eNOS) inhibitors, and NO donors. IL-1β was measured in cell culture supernatants. Patch-clamp and intracellular Ca(2+) imaging experiments were performed on HEK cells overexpressing human P2X7R or P2X7R with point mutations at cysteine residues in the cytoplasmic C-terminal domain. RESULTS: The inhibitory effect of nAChR agonists on the BzATP-induced IL-1β release was reversed in the presence of eNOS inhibitors (L-NIO, L-NAME) as well as in U937 cells after silencing of eNOS expression. In peripheral blood mononuclear leukocytes from eNOS gene-deficient mice, the inhibitory effect of nAChR agonists was absent, suggesting that nAChRs signal via eNOS to inhibit the BzATP-induced IL-1β release. Moreover, NO donors (SNAP, S-nitroso-N-acetyl-DL-penicillamine; SIN-1) inhibited the BzATP-induced IL-1β release by mononuclear phagocytes. The BzATP-induced ionotropic activity of the P2X7R was abolished in the presence of SIN-1 in both, Xenopus laevis oocytes and HEK cells over-expressing the human P2X7R. This inhibitory effect of SIN-1 was absent in HEK cells expressing P2X7R, in which C377 was mutated to alanine, indicating the importance of C377 for the regulation of the P2X7R function by protein modification. CONCLUSION: We provide first evidence that ion flux-independent, metabotropic signaling of monocytic nAChRs involves eNOS activation and P2X7R modification, resulting in an inhibition of ATP signaling and ATP-mediated IL-1β release. This signaling pathway might be an interesting target for the treatment of inflammatory disorders. Frontiers Media S.A. 2023-03-09 /pmc/articles/PMC10034071/ /pubmed/36969210 http://dx.doi.org/10.3389/fimmu.2023.1140592 Text en Copyright © 2023 Richter, Asci, Singh, Yakoob, Meixner, Zakrzewicz, Liese, Hecker, Wilker, Stumpf, Schlüter, Rohde, Gödecke, Padberg, Manzini, Schmalzing and Grau https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Richter, Katrin Asci, Nilay Singh, Vijay K. Yakoob, Sanaria Hawro Meixner, Marion Zakrzewicz, Anna Liese, Juliane Hecker, Andreas Wilker, Sigrid Stumpf, Sabine Schlüter, Klaus-Dieter Rohde, Marius Gödecke, Axel Padberg, Winfried Manzini, Ivan Schmalzing, Günther Grau, Veronika Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes |
title | Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes |
title_full | Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes |
title_fullStr | Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes |
title_full_unstemmed | Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes |
title_short | Activation of endothelial NO synthase and P2X7 receptor modification mediates the cholinergic control of ATP-induced interleukin-1β release by mononuclear phagocytes |
title_sort | activation of endothelial no synthase and p2x7 receptor modification mediates the cholinergic control of atp-induced interleukin-1β release by mononuclear phagocytes |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10034071/ https://www.ncbi.nlm.nih.gov/pubmed/36969210 http://dx.doi.org/10.3389/fimmu.2023.1140592 |
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