Phosphocholine-Modified Macromolecules and Canonical Nicotinic Agonists Inhibit ATP-Induced IL-1β Release This information is current as Andreas Hecker, Mira Küllmar, Sigrid Wilker, Katrin of September 24, 2021. Richter, Anna Zakrzewicz, Srebrena Atanasova, Verena Mathes, Thomas Timm, Sabrina Lerner, Jochen Klein, Andreas Kaufmann, Stefan Bauer, Winfried Padberg, Wolfgang Kummer, Sabina Janciauskiene, Martin Fronius, Elke K. H. Schweda, Günter Lochnit and Veronika Grau Downloaded from J Immunol 2015; 195:2325-2334; Prepublished online 22 July 2015; doi: 10.4049/jimmunol.1400974 http://www.jimmunol.org/content/195/5/2325 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2015/07/22/jimmunol.140097 Material 4.DCSupplemental References This article cites 42 articles, 11 of which you can access for free at: http://www.jimmunol.org/content/195/5/2325.full#ref-list-1 by guest on September 24, 2021 Why The JI? 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The Journal of Immunology Phosphocholine-Modified Macromolecules and Canonical Nicotinic Agonists Inhibit ATP-Induced IL-1b Release Andreas Hecker,*,1 Mira Kullmar,*€ ,1 Sigrid Wilker,* Katrin Richter,*,† Anna Zakrzewicz,* Srebrena Atanasova,* Verena Mathes,* Thomas Timm,‡ Sabrina Lerner,* Jochen Klein,x Andreas Kaufmann,{ Stefan Bauer,{,2 Winfried Padberg,*,2 Wolfgang Kummer,‖,2 Sabina Janciauskiene,#,2 Martin Fronius,†,** Elke K. H. Schweda,†† Gunter€ Lochnit,‡ and Veronika Grau*,2 IL-1b is a potent proinflammatory cytokine of the innate immune system that is involved in host defense against infection. However, increased production of IL-1b plays a pathogenic role in various inflammatory diseases, such as rheumatoid arthritis, gout, sepsis, stroke, and transplant rejection. To prevent detrimental collateral damage, IL-1b release is tightly controlled and Downloaded from typically requires two consecutive danger signals. LPS from Gram-negative bacteria is a prototypical first signal inducing pro–IL- 1b synthesis, whereas extracellular ATP is a typical second signal sensed by the ATP receptor P2X7 that triggers activation of the NLRP3-containing inflammasome, proteolytic cleavage of pro–IL-1b by caspase-1, and release of mature IL-1b. Mechanisms controlling IL-1b release, even in the presence of both danger signals, are needed to protect from collateral damage and are of therapeutic interest. In this article, we show that acetylcholine, choline, phosphocholine, phosphocholine-modified LPS from Haemophilus influenzae, and phosphocholine-modified protein efficiently inhibit ATP-mediated IL-1b release in human and rat http://www.jimmunol.org/ monocytes via nicotinic acetylcholine receptors containing subunits a7, a9, and/or a10. Of note, we identify receptors for phosphocholine-modified macromolecules that are synthesized by microbes and eukaryotic parasites and are well-known mod- ulators of the immune system. Our data suggest that an endogenous anti-inflammatory cholinergic control mechanism effectively controls ATP-mediated release of IL-1b and that the same mechanism is used by symbionts and misused by parasites to evade innate immune responses of the host. The Journal of Immunology, 2015, 195: 2325–2334. nterleukin-1b is a multifunctional proinflammatory cytokine high systemic levels of IL-1b cause fever, shock, and organ b of the innate immune system that plays an important role in damage. Thus, increased production and release of IL-1 play by guest on September 24, 2021 I host defense (1). However, excessive release is hazardous because a pathogenic role in various inflammatory diseases, such as rheumatoid arthritis, sepsis, stroke, and transplant rejection (2). *Laboratory of Experimental Surgery, Department of General and Thoracic Surgery, To prevent detrimental collateral damage, IL-1b release typi- Justus-Liebig-University Giessen, D-35385 Giessen, Germany; †Institute of Animal cally requires two consecutive danger signals (3, 4). LPS from Physiology, Justus-Liebig-University Giessen, D-35392 Giessen, Germany; ‡Protein Analytics, Institute of Biochemistry, Justus-Liebig-University Giessen, Gram-negative bacteria is a prototypicalfirstsignalinducing D-35392 Giessen, Germany; xDepartment of Pharmacology, Goethe University Col- pro–IL-1b synthesis. Extracellular ATP is a typical second { lege of Pharmacy, D-60438 Frankfurt, Germany; Institute for Immunology, Philipps- signal sensed by the ATP-receptor P2X7 that triggers activation University Marburg, D-35043 Marburg, Germany; ‖Institute of Anatomy and Cell Biology, Justus-Liebig-University Giessen, D-35392 Giessen, Germany; #Department of the NLRP3-containing inflammasome, proteolytic cleavage of Respiratory Medicine, Hannover Medical School, D-30625 Hannover, Germany; of pro–IL-1b by caspase-1, and release of mature IL-1b (3–5). **Department of Physiology, University of Otago, Dunedin 9054, New Zealand; and †† Mechanisms controlling IL-1b release are needed to protect the Division of Chemistry, Department of Physics, Chemistry and Biology, Linko¨ping University, S-58183 Linko¨ping, Sweden host from excessive inflammation but are largely unexplored. 1A.H. and M.K. contributed equally to this study. Acetylcholine (ACh) was the first neurotransmitter to be iden- 2S.B., W.P., W.K., S.J., and V.G. are members of the German Centre for Lung Research. tified, but it is also a regulator of the immune system with pro- and Received for publication April 16, 2014. Accepted for publication June 23, 2015. anti-inflammatory potential (6, 7). The first data pointing to an anti-inflammatory role of ACh date back to the year 2000, when This work was supported by a research grant from the University Medical Centre Giessen and Marburg (to A.H. and S.B.), the Landes-Offensive zur Entwicklung Borovikova et al. (8) demonstrated that vagus nerve stimulation Wissenschaftlich-O¨ konomischer Exzellenz of the State of Hesse (Nonneuronal Cho- attenuates the systemic release of TNF in response to LPS. Fur- linergic Systems to M.K., J.K., W.K., M.F., G.L., and V.G.), and by the German a Research Foundation (GR 1094/6-1 to V.G.). thermore, these investigators described the -conotoxin–sensitive b Address correspondence and reprint requests to Prof. Veronika Grau, Laboratory of inhibition of the secretion of TNF, IL-1 , IL-6, and IL-18 from Experimental Surgery, Justus-Liebig-University Giessen, Feulgen-Strasse 10-12, LPS-primed human macrophages in vitro (8). The current notion D-35385 Giessen, Germany. E-mail address: [email protected] prevails that vagal stimulation triggers neuronal release of nor- The online version of this article contains supplemental material. epinephrine from the spleen that stimulates the release of ACh Abbreviations used in this article: ACh, acetylcholine; BzATP, 29(39)-O-(4-benzoyl- from splenic memory T cells. The released ACh is sensed by benzoyl)ATP trieethylammonium salt; CHRNA7, nicotinic receptor for ACh contain- ing the a7 subunit; CHRNA9, nicotinic receptor for ACh containing the a9 subunit; splenic macrophages via nicotinic receptors for ACh containing CHRNA10, nicotinic receptor for ACh containing the a10 subunit; LDH, lactate the a7 subunit (CHRNA7) (7, 9), leading to changes in cytokine dehydrogenase; PC, phosphocholine; PC-BSA, PC-modified BSA; PC-LPS, PC- expression via the Jak2-STAT3 signaling pathway (7, 10). This modified LPS; siRNA, small interfering RNA. pathway is suggested to downmodulate LPS-induced expression Copyright Ó 2015 by The American Association of Immunologists, Inc. 0022-1767/15/$25.00 of pro–IL-1b but not its cleavage or release. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1400974 2326 CHOLINERGIC INHIBITION OF IL-1b RELEASE We demonstrated that activated monocytes accumulating in 5% CO2, in RPMI 1640 supplemented with 10% FCS (FCS gold) and blood vessels of rat renal grafts produce ACh during acute re- 2mML-glutamine (all from PAA Laboratories, Co¨lbe, Germany). 9 9 jection (11, 12). Because monocytes express nicotinic receptors Thereafter, 2 (3 )-O-(4-benzoylbenzoyl)ATP trieethylammonium salt (BzATP; 100 mM; Sigma-Aldrich, Taufkirchen, Germany) was added for ACh, endogenously produced ACh might act as an autocrine in the presence or absence of ACh chloride (10 or 100 mM), nicotine or paracrine regulator of innate immunity. Interestingly, nico- (100 mM), and cholinesterase from Electrophorus electricus (1 U/ml) tinic ACh receptors do not function as ligand gated ion chan- (all from Sigma-Aldrich). Supernatants were collected and stored at 220˚C nels in primary rat monocytes and alveolar macrophages but before cytokines and lactate dehydrogenase (LDH) were measured. rather as metabotropic receptors reducing ATP-induced intra- U937 cells cellular calcium signals (11, 13). This led