The Transcriptional Repressor BLIMP1 Curbs Host Defenses by Suppressing Expression of the Chemokine CCL8 This information is current as Martina Severa, Sabina A. Islam, Stephen N. Waggoner, of October 1, 2021. Zhaozhao Jiang, Nancy D. Kim, Glennice Ryan, Evelyn Kurt-Jones, Israel Charo, Daniel R. Caffrey, Victor L. Boyartchuk, Andrew D. Luster and Katherine A. Fitzgerald J Immunol 2014; 192:2291-2304; Prepublished online 29 January 2014; doi: 10.4049/jimmunol.1301799 Downloaded from http://www.jimmunol.org/content/192/5/2291 Supplementary http://www.jimmunol.org/content/suppl/2014/01/28/jimmunol.130179 http://www.jimmunol.org/ Material 9.DCSupplemental References This article cites 78 articles, 28 of which you can access for free at: http://www.jimmunol.org/content/192/5/2291.full#ref-list-1 Why The JI? Submit online. by guest on October 1, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • 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 © 2014 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology The Transcriptional Repressor BLIMP1 Curbs Host Defenses by Suppressing Expression of the Chemokine CCL8 Martina Severa,*,† Sabina A. Islam,‡ Stephen N. Waggoner,x Zhaozhao Jiang,* Nancy D. Kim,‡ Glennice Ryan,* Evelyn Kurt-Jones,* Israel Charo,{ Daniel R. Caffrey,* Victor L. Boyartchuk,‖ Andrew D. Luster,‡ and Katherine A. Fitzgerald*,‖ The transcriptional repressor B lymphocyte–induced maturation protein 1 (BLIMP1) is a master regulator of B and T cell differentiation. To examine the role of BLIMP1 in innate immunity, we used a conditional knockout (CKO) of Blimp1 in myeloid cells and found that Blimp1 CKO mice were protected from lethal infection induced by Listeria monocytogenes. Transcriptome analysis of Blimp1 CKO macrophages identified the murine chemokine (C-C motif) ligand 8, CCL8, as a direct target of Blimp1- mediated transcriptional repression in these cells. BLIMP1-deficient macrophages expressed elevated levels of Ccl8, and conse- quently Blimp1 CKO mice had higher levels of circulating CCL8, resulting in increased neutrophils in the peripheral blood, Downloaded from promoting a more aggressive antibacterial response. Mice lacking the Ccl8 gene were more susceptible to L. monocytogenes infection than were wild-type mice. Although CCL8 failed to recruit neutrophils directly, it was chemotactic for g/d T cells, and CCL8-responsive g/d T cells were enriched for IL-17F. Finally, CCL8-mediated enhanced clearance of L. monocytogenes was dependent on g/d T cells. Collectively, these data reveal an important role for BLIMP1 in modulating host defenses by suppressing expression of the chemokine CCL8. The Journal of Immunology, 2014, 192: 2291–2304. http://www.jimmunol.org/ lymphocyte–induced maturation protein 1 (BLIMP1) is differentiation, homeostasis, and/or function of both CD4 and a transcriptional repressor critical for early embryonic CD8 T cells (9, 10). It also plays a central role in the differenti- B development in multiple species (1–3). BLIMP1 was first ation and function of keratinocytes (11) and sebocytes (12) and in identified as a factor that bound to the positive regulatory domains the tolerogenic function of DCs (13). The role of BLIMP1 in I and III of the IFN-b enhancer, where it was shown to compete macrophages and antimicrobial defenses, however, has not been with IFN regulatory factors to attenuate IFN-b gene transcription examined in detail. (4, 5). BLIMP1 is best studied, however, in the context of terminal Macrophages are one of the major mediators of inflammation and critical effector cells of innate immunity. They serve as host differentiation of B lymphocytes to plasma cells (6–8) and in the by guest on October 1, 2021 cells for intracellular pathogens, including Listeria monocytogenes (14). L. monocytogenes is a Gram-positive facultative intracellular *Division of Infectious Diseases and Immunology, Department of Medicine, Univer- sity of Massachusetts Medical School, Worcester, MA 01605; †Dipartimento di bacterium, which causes disease in humans as a result of ingestion Malattie Infettive, Parassitarie ed Immuno-Mediata Malattie, Istituto Superiore di of contaminated foods. L. monocytogenes infections are generally ‡ Sanita`, 299-00161 Rome, Italy; Division of Rheumatology, Allergy and Immunol- limited; however, lethal infections can occur in immunocompro- ogy, Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02482; xDepartment of Pathology mised individuals, pregnant women, and neonates. Upon inges- and Program in Immunology and Virology, University of Massachusetts Medical tion, bacteria invade the intestinal epithelium, enter the draining School, Worcester, MA 01655; {Gladstone Institute of Cardiovascular Disease, Uni- versity of California, San Francisco, San Francisco CA 94158; and ‖Department of lymph node, and disseminate via the bloodstream to the liver and Cancer Research and Molecular Medicine, Centre of Molecular Inflammation Re- spleen (15). This rapid clearance of bacteria from the peripheral search, Norwegian University of Science and Technology, 7489 Trondheim, Norway blood is generally attributed to the resident liver macrophages— Received for publication July 11, 2013. Accepted for publication December 11, 2013. the Kupffer cells, which line the liver sinusoids—and to the This work was supported by the National Institutes of Health (Grants AI067497 to hepatocytes. Within the first 24 h, a coordinated interaction between K.A.F., R37AI040618 to A.D.L., and AI060991 to V.L.B.) and by Italy–USA Pro- neutrophils, g/d T cells, monocytes, and NK cells is instrumental in gram Grant 28C6 (to M.S.). preventing the bacterium from spreading and ensuring the survival The sequences presented in this article have been submitted to Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE53145) under ac- of the host. L. monocytogenes is an excellent model system to cession number GSE53145. evaluate the role of innate immunity in antimicrobial defenses. Address correspondence and reprint requests to Dr. Katherine A. Fitzgerald, Univer- Several classes of germline-encoded pattern recognition receptors sity of Massachusetts Medical School, Division of Infectious Diseases and Immu- (PRRs) have been implicated in the recognition of L. mono- nology, LRB Room 308, Department of Medicine, 364 Plantation Street, Worcester, MA 01605. E-mail address: kate.fi[email protected] cytogenes (14, 16). TLR-2 plays a key role in bacterial recognition The online version of this article contains supplemental material. at the cell surface (17–19). Upon escape of intracellular bacteria into the cytosol, L. monocytogenes engages nucleotide-binding olig- Abbreviations used in this article: BLIMP1, B lymphocyte–induced maturation pro- tein 1; BMDM, bone marrow–derived macrophage; ChIP, chromatin immunoprecip- omerization domain (NOD) 2, members of the NACHT-, LRR- and itation; CKO, conditional knockout; GO, gene ontology; hpi, h postinfection; IRF, pyrin-domain–containing protein family, as well as the cytosolic IFN regulatory factor; KO, knockout; LLO, listeriolysin O; MOI, multiplicity of infection; NOD, nucleotide-binding oligomerization domain; PEC, peritoneal exu- DNA sensor AIM2 (absent in melanoma 2), all of which play date cell; PRR, pattern recognition receptor; qPCR, quantitative real-time RT-PCR; a role in the recognition of intracellular L. monocytogenes and RIP2, receptor-interacting serine-threonine kinase 2; TSB, tryptic soy broth; unr, release of IL-1b through the activation of caspase-1 (20–23). In unrelated; WT, wild-type. addition, L. monocytogenes triggers type I IFN gene transcription via Copyright Ó 2014 by The American Association of Immunologists, Inc. 0022-1767/14/$16.00 STING (stimulator of IFN genes) (24–28), TBK1 (TANK-binding www.jimmunol.org/cgi/doi/10.4049/jimmunol.1301799 2292 BLIMP1-REGULATED CCL8 EXPRESSION IN HOST DEFENSE kinase 1), and IFN regulatory factor (IRF) 3 (29–31). Bacterial peritoneal cavities with 5 ml ice-cold PBS 3 d after i.p. injection with 3 ml DNA and the second messenger cyclic-di-AMP represent the L. thioglycollate (Remel, Lenexa, KS). Splenocytes were isolated by disrupt- monocytogenes products driving these responses (31–33). ing aseptically harvested spleens in RPMI 1640 10% FCS, using a 70-mm pore size mesh cell strainer (BD Biosciences) to obtain single-cell sus- In this study, we examined the role of BLIMP1 in regulating pensions. Whole-blood samples were obtained by cardiac puncture using the innate immune response using L. monocytogenes as a model 1-ml syringes containing EDTA and from tails in sterile 5-ml polypropylene system. We found that Blimp1 gene expression was rapidly in- tubes containing PBS with 10 mM EDTA. Lysis of RBCs was performed duced by this bacterium in macrophages