Downloaded from http://www.jimmunol.org/ by guest on October 1, 2021 is online at: average * The Journal of Immunology , 15 of which you can access for free at: 2013; 190:531-538; Prepublished online 17 from submission to initial decision 4 weeks from acceptance to publication December 2012; doi: 10.4049/jimmunol.1201576 http://www.jimmunol.org/content/190/2/531 IL-33 Induces a Hyporesponsive Phenotype in Human and Mouse Mast Cells Mi-Yeon Jung, Daniel Smrz, Avanti Desai, Geethani Bandara, Tomonobu Ito, Shoko Iwaki, Jeong-Han Kang, Marcus V. Andrade, Susana C. Hilderbrand, Jared M. Brown, Michael A. Beaven, Dean D. Metcalfe and Alasdair M. Gilfillan J Immunol cites 33 articles Submit online. Every submission reviewed by practicing scientists ? is published twice each month by Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://jimmunol.org/subscription http://www.jimmunol.org/content/suppl/2012/12/17/jimmunol.120157 6.DC1 This article http://www.jimmunol.org/content/190/2/531.full#ref-list-1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material References Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of October 1, 2021. The Journal of Immunology IL-33 Induces a Hyporesponsive Phenotype in Human and Mouse Mast Cells Mi-Yeon Jung,* Daniel Smrz,*ˇ Avanti Desai,* Geethani Bandara,* Tomonobu Ito,* Shoko Iwaki,† Jeong-Han Kang,‡ Marcus V. Andrade,x Susana C. Hilderbrand,{ Jared M. Brown,{ Michael A. Beaven,† Dean D. Metcalfe,* and Alasdair M. Gilfillan* IL-33 is elevated in afflicted tissues of patients with mast cell (MC)–dependent chronic allergic diseases. Based on its acute effects on mouse MCs, IL-33 is thought to play a role in the pathogenesis of allergic disease through MC activation. However, the mani- festations of prolonged IL-33 exposure on human MC function, which best reflect the conditions associated with chronic allergic disease, are unknown. In this study, we found that long-term exposure of human and mouse MCs to IL-33 results in a substantial reduction of MC activation in response to Ag. This reduction required >72 h exposure to IL-33 for onset and 1–2 wk for reversion following IL-33 removal. This hyporesponsive phenotype was determined to be a consequence of MyD88-dependent attenuation of Downloaded from signaling processes necessary for MC activation, including Ag-mediated calcium mobilization and cytoskeletal reorganization, potentially as a consequence of downregulation of the expression of phospholipase Cg1 and Hck. These findings suggest that IL-33 may play a protective, rather than a causative, role in MC activation under chronic conditions and, furthermore, reveal regulated plasticity in the MC activation phenotype. The ability to downregulate MC activation in this manner may provide alternative approaches for treatment of MC-driven disease. The Journal of Immunology, 2013, 190: 531–538. http://www.jimmunol.org/ ast cells (MCs) are tissue-resident cells of hematopoietic MC activation (6–9), it has been proposed that IL-33 may play origin that participate in innate and acquired immune a role in the pathogenesis of allergic disorders. The effects of M defense mechanisms through the release of an array of IL-33 on MC activation have, however, been controversial. Al- inflammatory mediators following receptor-dependent activation though it is recognized that IL-33 induces cytokine release from (1). These mediators, however, are also responsible for initiating the MCs in culture (7, 10–12) and markedly enhances the ability of cellular and pathological manifestations of allergic disorders, in- Ag to elicit this response (6–9), some studies report that IL-33 cluding anaphylaxis, asthma, rhinitis, and atopic dermatitis (2). MCs also stimulates or enhances degranulation and production of are predominantly activated through Ag/IgE-mediated aggregation PGD2 (9, 12), whereas others report that IL-33 neither induces by guest on October 1, 2021 of the high-affinity IgE receptor FcεRI (3). However, it is apparent or enhances FcεRI-mediated degranulation and eicosanoid pro- that cytokines and other molecules present in the surrounding mi- duction (7, 8, 10, 11). lieu can acutely amplify MC signaling and responses to Ag (4). The amplification of cytokine release by IL-33 in Ag-stimulated IL-33 levels are reported to be elevated in lungs of patients with MCs in culture was noted after acute exposure to IL-33, conditions chronic asthma (5) and, based on the reports that IL-33 enhances that may not mimic the long-term effects of IL-33 on MC function associated with pathologic conditions of atopic or rheumatoid arthritic disease. We thus investigated the long-term effects of *Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Dis- IL-33 on the ability of Ag to activate human and mouse MCs. eases, National Institutes of Health, Bethesda, MD 20892; †Laboratory of Molecular Immunology, National Heart, Lung, and Blood Institute, National Institutes of In this study, we report that, following prolonged IL-33 exposure, Health, Bethesda, MD 20892; ‡Laboratory of Cell Biology, National Cancer Institute, MCs become refractory to stimulation via FcεRI, with respect to x National Institutes of Health, Bethesda, MD 20892; Department of Internal Medi- degranulation, as a consequence of the attenuation of critical sig- cine, School of Medicine, Federal University of Minas Gerais, Belo Horizonte, MG 30130-100, Brazil; and {Department of Pharmacology and Toxicology, Brody School naling processes required for MC activation. To our knowledge, these of Medicine, East Carolina University, Greenville, NC 27834 findings provide the first evidence that human (hu)MCs can be re- Received for publication June 7, 2012. Accepted for publication November 6, 2012. programmed to a hyporesponsive phenotype in this manner and thus This work was supported by the Division of Intramural Research of the National may have important implications for how MC activation may be Institute of Allergy and Infectious Diseases, the National Heart, Lung, and Blood regulated in health and disease. The ability to manipulate MC acti- Institute, and the National Cancer Institute within the National Institutes of Health, National Institutes of Health Grants R01 ES019311 (to J.M.B.) and R01 TW006612 vation in this manner may also provide a basis for the development of (Global Research Initiative Program for New Foreign Investigators) (to M.V.A.), as novel approaches for therapeutic intervention of MC-driven disease. well as by the Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´gico and the Fundac¸a˜o de Amparo a` Pesquisa do Estado de Minas Gerais, Brazil (to M.V.A.). Materials and Methods The sequences presented in this article have been submitted to the Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo/) under accession number GSE39382. Cell culture Address correspondence and reprint requests to Dr. Alasdair M. Gilfillan, Laboratory Primary huMCs were prepared from CD34+ peripheral blood progenitors of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National isolated from healthy volunteers following informed consent under a Institutes of Health, 10 Center Drive, MSC 1881, Building 10, Room 11C206, Be- protocol (NCT00001756) approved by the National Institutes of Health thesda, MD 20892. E-mail address: agilfi[email protected] Internal Review Board. Human MCs were prepared and cultured as de- The online version of this article contains supplemental material. scribed (13) with or without recombinant human IL-33 (10 ng/ml) and Abbreviations used in this article: BMMC, bone marrow–derived mast cell; HSA, used for studies between 7 and 9 wk age. human serum albumin; hu, human; MC, mast cell; MFI, mean fluorescence intensity; All mouse studies, with the exception of those involving the use of 2 2 PLC, phospholipase C; SCF, stem cell factor; WT, wild-type. ST2 / mice, were conducted under a protocol approved by the National www.jimmunol.org/cgi/doi/10.4049/jimmunol.1201576 532 IL-33 PROGRAMS THE MAST CELL ACTIVATION PHENOTYPE Institute of Allergy and Infectious Diseases Institutional Animal Care In vivo studies and isolation of peritoneal MCs and Use Committee at the National Institutes of Health. Studies involving the use of ST22/2 mice were approved by East Carolina University’s In- The acute effects of IL-33 (IL-33–induced anaphylaxis) were examined in stitutional Animal Care and Use Committee. These studies were conducted 6 wk old C57BL/6 mice (The Jackson Laboratory). The mice were sen- in accordance with the National Institutes of Health Guidelines for the sitized with anti-DNP IgE mAb (21) (3 mg i.v.), a gift from Juan Rivera Care and Use of Laboratory Animals. (National Institute of Arthritis and Musculoskeletal and Skin Diseases, Mouse bone marrow–derived MCs (BMMCs) were prepared from National Institutes of Health), and isolated from ascites supplied by wild-type (WT; C57BL/6 background; The Jackson Laboratory, Bar Dr. Fu-Tong Liu (Davis School of Medicine, University of California, Harbor, ME), ST22/2,andMyD882/2 mice. ST22/2 mice on a C57BL/6 Sacramento, CA). After 24 h, the mice were injected with rIL-33 (2 mg genetic background were obtained from Dr. Robert B. Fick at Merck in 200 ml) or PBS retro-orbitally (i.v), and the anaphylactic response Research Laboratories (Division of Biologics, Palo Alto, CA). Homo- was monitored by recording changes in core body temperature every zygous MyD882/2 mice on a C57BL/6 genetic background (14, 15) 5 min for 2 h using an implantable electronic transponder (IPTT-300; were obtained from Dr.