Novel Resolvin D2 Receptor Axis in Infectious Inflammation Nan Chiang, Xavier de la Rosa, Stephania Libreros and Charles N. Serhan This information is current as of September 25, 2021. J Immunol 2017; 198:842-851; Prepublished online 19 December 2016; doi: 10.4049/jimmunol.1601650 http://www.jimmunol.org/content/198/2/842 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2016/12/16/jimmunol.160165 Material 0.DCSupplemental References This article cites 51 articles, 19 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/198/2/842.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 25, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2017 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Novel Resolvin D2 Receptor Axis in Infectious Inflammation Nan Chiang,1 Xavier de la Rosa,1 Stephania Libreros, and Charles N. Serhan Resolution of acute inflammation is an active process governed by specialized proresolving mediators, including resolvin (Rv)D2, that activates a cell surface G protein–coupled receptor, GPR18/DRV2. In this study, we investigated RvD2-DRV2–dependent resolution mechanisms using DRV2-deficient mice (DRV2-knockout [KO]). In polymicrobial sepsis initiated by cecal ligation and puncture, RvD2 (∼2.7 nmol/mouse) significantly increased survival (>50%) of wild-type mice and reduced hypothermia and bacterial titers compared with vehicle-treated cecal ligation and puncture mice that succumbed at 48 h. Protection by RvD2 was abolished in DRV2-KO mice. Mass spectrometry–based lipid mediator metabololipidomics demonstrated that DRV2-KO infectious exudates gave higher proinflammatory leukotriene B4 and procoagulating thromboxane B2, as well as lower specialized proresolving mediators, including RvD1 and RvD3, compared with wild-type. RvD2-DRV2–initiated intracellular signals were investigated using mass cytometry (cytometry by time-of-flight), which demonstrated that RvD2 enhanced phosphorylation of CREB, ERK1/2, and STAT3 in WT but not DRV2-KO macrophages. Monitored by real-time imaging, RvD2–DRV2 interaction Downloaded from significantly enhanced phagocytosis of live Escherichia coli, an action dependent on protein kinase A and STAT3 in macrophages. Taken together, we identified an RvD2/DRV2 axis that activates intracellular signaling pathways that increase phagocytosis- mediated bacterial clearance, survival, and organ protection. Moreover, these results provide evidence for RvD2-DRV2 and their downstream pathways in pathophysiology of infectious inflammation. The Journal of Immunology, 2017, 198: 842–851. nflammation is a protective response to defend the host conserved chemical structures derived from polyunsaturated fatty http://www.jimmunol.org/ against infection and injury (1). Ungoverned and excessive acids, including eicosapentaenoic acid–derived E-series resolvins I inflammation is an underlying pathology of many prevalent and docosahexaenoic acid (DHA)–derived D-series resolvins, diseases, including cardiovascular diseases, diabetes, arthritis, and protectins, and maresins. Complete stereochemistries of these sepsis (2–4). Complete resolution of acute inflammatory responses SPM are established and total organic synthesis achieved that also was thought to be a passive process with dissipation or dilution of confirmed their potent proresolving actions. SPM are potent me- local chemoattractants and proinflammatory mediators, allowing diators governing not only innate immune responses in host de- tissues to return to homeostasis (1). In recent years, we obtained fense, but also pain, organ protection, and tissue remodeling (recently the first evidence, to our knowledge, that resolution of self-limited reviewed in Ref. 8). inflammation is not merely a passive termination, but rather an SPM derived from DHA including resolvin (Rv)D2 (7S,16R,17S- by guest on September 25, 2021 actively orchestrated programmed response that is rapidly turned trihydroxy-docosa-4Z,8E,10Z,12E,14E,19Z-hexaenoic acid) were on during acute inflammatory challenges, permitting inflamed and first identified and isolated from murine self-resolving exudates injured tissues to return via catabasis to function (5, 6). This event during the resolution phase of self-limited acute inflammation is driven in part by temporal lipid mediator (LM) class switching in vivo (12). The biosynthesis of RvD2 involves 17-lipoxygenation from generation of proinflammatory mediators (e.g., leukotriene of DHA to 17S-hydroperoxy-DHA that is further transformed en- [LT]B4) to the biosynthesis of lipoxins (LX) (7) and specialized zymatically to a 7(8)epoxide-containing intermediate in leukocytes proresolving mediators (SPM) (8–11). SPM are evolutionally via 5-lipoxygenase, followed by enzymatic hydrolysis to form RvD2. Endogenous RvD2 production is documented in human serum, plasma (13), adipose tissue (14), placenta (15), lung (16), Center for Experimental Therapeutics and Reperfusion Injury, Department of Anes- breast milk (17), and sepsis patients (18). With isolated human thesiology, Perioperative and Pain Medicine, Harvard Institutes of Medicine, Brig- polymorphonuclear neutrophils (PMN), RvD2 increases intracel- ham and Women’s Hospital and Harvard Medical School, Boston, MA 02115 lular phagosomal reactive oxygen species generation for microbial 1 N.C. and X.d.l.R. contributed equally to this work. killing (19). In whole blood at a single cell level using microfluidic ORCID: 0000-0003-4627-8545 (C.N.S.). chambers, RvD2 limits PMN chemotaxis and direct travel as well Received for publication September 22, 2016. Accepted for publication November as increases random movement toward an IL-8 chemotactic gradi- 15, 2016. ent (20). RvD2 also decreases monocyte adhesion to adipocytes as This work was supported in part by National Institutes of Health Grants R01 well as their transadipose migration (14). RvD2 is a potent immu- GM38765 (to C.N.S.) and R01 GM38765-29S1 (to S.L.). noresolvent that stereoselectively reduces excessive PMN trafficking Address correspondence and reprint requests to Prof. Charles N. Serhan, Brigham and Women’s Hospital, Building for Transformative Medicine, Suite 3-016, 60 Fen- in peritonitis and improves survival in sepsis (19). RvD2’s potent wood Road, Boston, MA 02115. E-mail address: [email protected] nanogram actions are also protective in disease models where RvD2 The online version of this article contains supplemental material. prevents inflammatory bowel disease such as colitis (21), alleviates Abbreviations used in this article: BMDM, bone marrow–derived macrophage; CLP, inflammatory and fibromyalgia-induced pain (22, 23), increases cecal ligation and puncture; CyTOF, cytometry by time-of-flight; DHA, docosahexaenoic survival following burn wound and reduces kidney and liver injuries acid; KO, knockout; LM, lipid mediator; LT, leukotriene; LX, lipoxin; MMP, matrix metalloproteinase; MRM, multiple reaction monitoring; PKA, protein kinase A; PMN, in mice (24, 25), and reduces periodontitis (26) as well as nerve polymorphonuclear neutrophil; Rv, resolvin; RvD2, resolvin D2 (7S,16R,17S-trihydroxy- injuries as seen in Parkinson disease (27). docosa-4Z,8E,10Z,12E,14E,19Z-hexaenoic acid); SPM, specialized proresolving media- Resolution at the cellular level consists of cessation of PMN tor; STZ, serum-treated zymosan particle; TX, thromboxane; WT, wild-type. entry into the tissue and elevated efferocytosis (i.e., macrophage Copyright Ó 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$30.00 phagocytosis of apoptotic PMN) (1). We introduced a quantitative www.jimmunol.org/cgi/doi/10.4049/jimmunol.1601650 The Journal of Immunology 843 definition of resolution of self-limited sterile acute inflammation d4-5S-HETE, d4-PGE2, and d5-RvD2 internal standards (500 pg each) denoted resolution indices that permits assessment of the resolu- were added to facilitate quantification. All samples were kept at 220˚C for tion properties of SPM and pinpoints their unique mechanisms of 45 min to allow protein precipitation and then subjected to solid-phase extraction as described (13). Extracted samples were analyzed by a liquid action (6). SPM each lower the magnitude of leukocyte infiltration chromatography–tandem mass spectrometry system (QTRAP 5500; AB and/or shorten the resolution interval, which is the interval from Sciex) equipped with an LC-20AD HPLC (Shimadzu, Tokyo, Japan). A the time point of maximum PMN infiltration to the time point of Poroshell 120 EC-18 column (100 mm 3 4.6 mm 3 2.7 mm; Agilent 50% PMN reduction in peritoneal exudates (17, 28). We also Technologies, Santa Clara, CA) was kept in a column oven maintained at 50˚C, and LM were eluted with a gradient of methanol/water/acetic acid assessed the actions of SPM in resolution of