Inhibition of Histone Deacetylases Permits Lipopolysaccharide-Mediated Secretion of Bioactive IL-1β via a Caspase-1−Independent Mechanism This information is current as of September 28, 2021. Dominik Stammler, Tatjana Eigenbrod, Sarah Menz, Julia S. Frick, Matthew J. Sweet, Melanie R. Shakespear, Jonathan Jantsch, Isabel Siegert, Sabine Wölfle, Julian D. Langer, Ina Oehme, Liliana Schaefer, Andre Fischer, Judith Knievel, Klaus Heeg, Alexander H. Dalpke and Konrad A. Bode Downloaded from J Immunol published online 30 October 2015 http://www.jimmunol.org/content/early/2015/10/30/jimmun ol.1501195 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2015/10/30/jimmunol.150119 Material 5.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 28, 2021 • No Triage! 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Published October 30, 2015, doi:10.4049/jimmunol.1501195 The Journal of Immunology Inhibition of Histone Deacetylases Permits Lipopolysaccharide-Mediated Secretion of Bioactive IL-1b via a Caspase-1–Independent Mechanism Dominik Stammler,* Tatjana Eigenbrod,* Sarah Menz,† Julia S. Frick,† Matthew J. Sweet,‡ Melanie R. Shakespear,‡ Jonathan Jantsch,x,{ Isabel Siegert,{ Sabine Wo¨lfle,* Julian D. Langer,‖ Ina Oehme,# Liliana Schaefer,** Andre Fischer,†† Judith Knievel,‡‡ Klaus Heeg,* Alexander H. Dalpke,* and Konrad A. Bode* Histone deacetylase (HDAC) inhibitors (HDACi) are clinically approved anticancer drugs that have important immune- modulatory properties. We report the surprising finding that HDACi promote LPS-induced IL-1b processing and secretion in Downloaded from human and murine dendritic cells and murine macrophages. HDACi/LPS-induced IL-1b maturation and secretion kinetics differed completely from those observed upon inflammasome activation. Moreover, this pathway of IL-1b secretion was depen- dent on caspase-8 but was independent of the inflammasome components NACHT, LRR, and PYD domains-containing protein 3, apoptosis-associated speck-like protein containing a carboxyl-terminal caspase-recruitment domain, and caspase-1. Genetic stud- ies excluded HDAC6 and HDAC10 as relevant HDAC targets in this pathway, whereas pharmacological inhibitor studies impli- cated the involvement of HDAC11. Treatment of mice with HDACi in a dextran sodium sulfate–induced colitis model resulted in a http://www.jimmunol.org/ strong increase in intestinal IL-1b, confirming that this pathway is also operative in vivo. Thus, in addition to the conventional inflammasome-dependent IL-1b cleavage pathway, dendritic cells and macrophages are capable of generating, secreting, and processing bioactive IL-1b by a novel, caspase-8–dependent mechanism. Given the widespread interest in the therapeutic target- ing of IL-1b, as well as the use of HDACi for anti-inflammatory applications, these findings have substantial clinical implications. The Journal of Immunology, 2015, 195: 000–000. ppropriate regulation of local immunity is a prerequisite APC, such as macrophages and dendritic cells (DC) respond, for the maintenance of local tissue homeostasis, and depending on the stimulus, with the secretion of several in- A dysregulated innate immunity can result in acute or flammatory cytokines, including TNF-a, IL-12p40, IL-6, and by guest on September 28, 2021 chronic inflammation. Both hematopoietic and nonhematopoietic IL-1b (1). cells of the innate immune system recognize pathogen- and The IL-1b cytokine exhibits diverse biological activities and damage-associated molecular patterns via a variety of pattern- plays a central role in acute and chronic inflammation. Because of recognition receptors, including the TLR, nucleotide-binding oligo- its potent proinflammatory effects, IL-1b production is tightly merization domain protein-like receptors (NLR), such as nucleotide- regulated. At least two stimuli are needed for the secretion of binding oligomerization domain-containing protein (NOD)1 and bioactive IL-1b: a first stimulus (e.g., a TLR ligand) is required for NOD2 and the inflammasome-associated NLRP/NLRC sensor inducible expression of pro–IL-1b and priming of the inflamma- proteins, C-type lectin receptors, and Rig-I–like receptors (1). some, after which a second signaling event is necessary for pro–IL- Stimulation of these receptors results in cellular activation and 1b processing and the secretion of active IL-1b (2, 3). Pro–IL-1b the tailored release of a panel of cytokines and chemokines that itself is biologically inactive and requires proteolytic cleavage to is appropriate to the class of pathogen encountered. Professional enable secretion and biological responses. Classically, pro–IL-1b is *Department of Infectious Diseases, Medical Microbiology and Hygiene, Heidelberg Address correspondence and reprint requests to Dr. Konrad A. Bode, Department of University Hospital, Heidelberg 69120, Germany; †Institute of Medical Microbiology Infectious Diseases, Medical Microbiology and Hygiene, Heidelberg University Hos- and Hygiene, University of Tubingen,€ Tubingen€ 70276, Germany; ‡Institute for Mo- pital, Im Neuenheimer Feld 324, Heidelberg 69120, Germany. E-mail address: lecular Bioscience, University of Queensland, Brisbane, St. Lucia, Queensland 4072, [email protected] Australia; xInstitute of Clinical Microbiology and Hygiene, University Hospital Regens- { The online version of this article contains supplemental material. burg, Regensburg 93053, Germany; Institute of Microbiology, University of Erlangen- Nuremberg, Erlangen 91054, Germany; ‖Department of Molecular Membrane Biology, Abbreviations used in this article: ASC, apoptosis-associated speck-like protein con- Max-Planck-Institute for Biophysics, Frankfurt 60438, Germany; #Clinical Cooperation taining a carboxyl-terminal caspase-recruitment domain; BMDC, bone marrow–de- Unit Pediatric Oncology, German Cancer Research Center, Heidelberg 69120, Ger- rived DC; BMDM, BM-derived macrophage; DC, dendritic cell; DSS, dextran many; **Institute of Pharmacology, Goethe University Frankfurt, Frankfurt 60590, sodium sulfate; HATi, histone acetyltransferase inhibitor; HDAC, histone deacety- Germany; ††German Center for Neurodegenerative Diseases, Department of Psychiatry lase; HDACi, HDAC inhibitor; IQ, imiquimod; KO, knockout; LTA, lipoteichoic and Psychotherapy, University Medical Center Go¨ttingen, Go¨ttingen 37077, Germany; acid; MDP, muramyl dipeptide; NLR, nucleotide-binding oligomerization domain and ‡‡Department of Gastroenterology, Hepatology, and Infectious Disease, University protein-like receptor; NLRP3, NACHT, LRR, and PYD domains-containing protein Hospital, Heinrich Heine University of Dusseldorf,€ Dusseldorf€ 40225, Germany 3; NOD, nucleotide-binding oligomerization domain-containing protein; PD106, pimelic diphenylamide 106; p(I:C), poly(deoxyinosinic-deoxycytidylic) acid; poly ORCIDs: 0000-0003-0974-7402 (J.K.); 0000-0001-9085-965X (K.A.B.). (I:C), polyinosinic-polycytidylic acid; SAHA, suberanilohydroxamic acid; siRNA, Received for publication June 3, 2015. Accepted for publication October 5, 2015. small interfering RNA; TRIF, TIR domain-containing adapter-inducing IFN-b; TSA, trichostatin A; WT, wild-type. This work was supported by German Research Foundation Grants BO 3673/1-1 (to K.A.B.) and INST 114089/4-1 FUGG (to K.H.). M.J.S. is supported by a National Ó Health and Medical Research Council of Australia Senior Research Fellowship Copyright 2015 by The American Association of Immunologists, Inc. 0022-1767/15/$25.00 (APP1003470). www.jimmunol.org/cgi/doi/10.4049/jimmunol.1501195 2 HDACi PERMITS LPS-MEDIATED SECRETION OF BIOACTIVE IL-1b cleaved by caspase-1. Stimulation of primed cells with damage- Cell Signaling Technology (Danvers, MA). Acetylated histone H3 was associated molecular patterns, such as ATP and urate crystals, or detected by an Ab purchased from Active Motif (Carlsbad, CA). The Ab pore-forming toxins, like nigericin, results in the assembly of a against b-actin was from Upstate Biotechnologies (Lake Placid, NY). MyD88-, apoptosis-associated speck-like protein containing a carboxy- large protein complex, the so-called “inflammasome,” which is terminal caspase-recruitment domain (ASC)-, NACHT, LRR, and PYD responsible for activation of caspase-1. Despite a predominant role domains-containing protein 3 (NLRP3)-, HDAC6-, cathepsin B–, cathep- for caspase-1 in pro–IL-1b cleavage, other enzymes, such as sin S–, granzyme A–, granzyme B–, IL-1b–, and caspase-1/11–knockout neutrophil elastase (4), proteinase 3 (5), chymase, cathepsin G (6), (KO) mice were described (18–27); lps2 mutant strains carry a mutation in TIR domain-containing adapter-inducing IFN-b (TRIF) (28). HDAC10- granzyme A (7), the matrix metalloproteinases stromelysin 1, KO mice were purchased from The Jackson Laboratory