Statins Induce Regulatory T Cell Recruitment via a CCL1 Dependent Pathway Emilia Mira, Beatriz León, Domingo F. Barber, Sonia Jiménez-Baranda, Iñigo Goya, Luis Almonacid, Gabriel This information is current as Márquez, Angel Zaballos, Carlos Martínez-A., Jens V. Stein, of September 26, 2021. Carlos Ardavín and Santos Mañes J Immunol 2008; 181:3524-3534; ; doi: 10.4049/jimmunol.181.5.3524 http://www.jimmunol.org/content/181/5/3524 Downloaded from References This article cites 52 articles, 27 of which you can access for free at: http://www.jimmunol.org/content/181/5/3524.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • 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 by guest on September 26, 2021 *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 © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Statins Induce Regulatory T Cell Recruitment via a CCL1 Dependent Pathway1 Emilia Mira,* Beatriz Leo´n,* Domingo F. Barber,* Sonia Jime´nez-Baranda,* In˜igo Goya,* Luis Almonacid,* Gabriel Ma´rquez,† Angel Zaballos,* Carlos Martı´nez-A.,* Jens V. Stein,‡ Carlos Ardavı´n,* and Santos Man˜es2* The statins, a group of inhibitors of the 3-hydroxy-3-methylglutaryl coenzyme A reductase, are reported to influence a variety of immune system activities through 3-hydroxy-3-methylglutaryl coenzyme A reductase-dependent and -independent mechanisms. How statin treatment regulates immune system function in vivo nonetheless remains to be fully defined. We analyzed the immu- nomodulatory effects of lovastatin in a Candida albicans-induced delayed-type hypersensitivity reaction in mice. In this model, lovastatin administration reduced the acute inflammatory response elicited by C. albicans challenge. This anti-inflammatory activity of lovastatin was associated with a shift from a Th1 to a Th2 immune response, as well as an increase in the percentage Downloaded from of regulatory T cells at the inflammation site and in the regional draining lymph node. The lovastatin-induced increase in regulatory T cells in the inflamed skin was dependent on expression of CCL1, a chemokine that is locally up-regulated by statin administration. The anti-inflammatory effect of lovastatin was abrogated in CCL1-deficient mice. These results suggest that local regulation of chemokine expression may be an important process in statin-induced modulation of the immune system. The Journal of Immunology, 2008, 181: 3524–3534. http://www.jimmunol.org/ he statins are used extensively in medical practice be- mokines, master cytokines for T cell polarization, metallopro- cause of their ability to reduce cardiovascular mortality teases, and costimulatory and MHC molecules (reviewed in Refs. T and stroke (1). Although this protective activity was ini- 16 and 17). At high in vitro concentrations, statins might interfere tially ascribed to inhibition of cholesterol biosynthesis, it is now with lymphocyte function by altering signaling in cholesterol-en- evident that statins are pleiotropic drugs with immunomodulatory riched lipid rafts (18, 19). and anti-inflammatory properties. Studies in animal models, as Statins are also reported to induce immunomodulation by 3-hy- well as epidemiological and small-scale clinical trials, suggest that droxy-3-methylglutaryl coenzyme A reductase-independent mech- ␤ statin pleiotropism might be clinically relevant in other patholo- anisms. Several statins bind to the 2 integrin LFA-1, preventing by guest on September 26, 2021 gies, including infection and sepsis (2–5), autoimmunity (6–9), its interaction with the ICAM-1 (20). Statin blockade of the LFA- and organ transplantation (10, 11). 1/ICAM-1 interaction has functional implications in T cell activa- The immunomodulatory effect of statins is thought to involve tion, leukocyte diapedesis, and HIV-1 infection (20–22). several mechanisms that act on different cell types (6, 12, 13). The extent to which each of these pleiotropic activities of statins Statins not only prevent de novo synthesis of cholesterol, they also contributes to immunomodulation in vivo is not well understood. decrease the pool of intermediate isoprenoid metabolites, such as Moreover, the reported immunomodulatory activities of statins farnesyl pyrophosphate or geranylgeranyl pyrophosphate, thus af- would be expected to attenuate the immune response. Paradoxi- fecting GTP-binding signaling proteins, including Ras and Rho- cally, the capacity to mount protective immune responses to intra- GTPases (2, 14, 15). Impairment of GTPase function allows statins cellular pathogens is not diminished in statin-treated patients. In- to affect actin-based cytoskeletal remodeling, cell motility, and the deed, regular statin use has been associated with reduced risk of activity of transcription factors that control the expression of che- bacteremia and sepsis (4, 5), as well as lower incidence and pro- gression of specific cancer types (23). We analyzed the in vivo effect of lovastatin in a murine model *Department of Immunology and Oncology, Centro Nacional de Biotecnologı´a/Con- of skin delayed-type hypersensitivity (DTH)3 response to heat- † sejo Superior de Investigaciones Cientificas (CSIC), Madrid, Spain; Genetrix, Ma- inactivated Candida albicans. Skin DTH responses involve Ag drid, Spain; and ‡Theodor Kocher Institute, University of Bern, Bern, Switzerland uptake, processing, and transport by APC to tissue-draining lymph Received for publication February 29, 2008. Accepted for publication June 20, 2008. nodes, where Ag-specific T cells are triggered to initiate a primary The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance immune response. Secondary exposure to the Ag results in a robust with 18 U.S.C. Section 1734 solely to indicate this fact. local inflammatory response. The coordinated cell and cytokine 1 This work was supported in part by the Spanish Ministry of Education and Science responses required to induce DTH make this experimental system (SAF2005-00241 and NAN2004-08805-C04/04 to S.M.; SAF2006-07609 to C.A.), ideal for in vivo analysis of the immune response. Fundacio´n para la Investigacio´n y Prevencio´n del SIDA en Espan˜a (36550/06 to S.M.), La Marato TV3 Foundation (to S.M.), the Comunidad de Madrid (DIFHE- Our data show that lovastatin reduced the inflammatory re- MAT-CM S-SAL-0304-2006 to S.M. and C.A.), and the European Union FP6 sponse to secondary Ag exposure, and that this effect was not a (INNOCHEM, FP6 LSHB-CT-2005-518167 to S.M. and C.M.A.). The Department of Immunology and Oncology was founded and is supported by the Spanish National Research Council and by Pfizer. 3 Abbreviations used in this paper: DTH, delayed-type hypersensitivity; Ct, cycle 2 Address correspondence and reprint requests to Dr. Santos Man˜es, Department of threshold; DC, dendritic cell; PO-LN, popliteal lymph node; SLO, secondary lym- Immunology and Oncology, Centro Nacional de Biotecnologı´a/Spanish National Re- phoid organs; Treg, regulatory T; WT, wild type. search Council, Darwin 3, Madrid E-28049 Spain. E-mail address: smanes@ cnb.csic.es Copyright © 2008 by The American Association of Immunologists, Inc. 0022-1767/08/$2.00 www.jimmunol.org The Journal of Immunology 3525 Table I. Changes in mRNA levels in the whole footpad after A 4.4 Kb 4.4 Kb C. albicans challenge of lovastatin- and vehicle-treated mice 6.4 Kb SacI SacI BamHI SphI NcoI NcoI BamHI SacI Lovastatin Lovastatin/Vehicle CCL1 WT SphI Molecule (2Ϫ⌬⌬Ct)a Vehicle (2Ϫ⌬⌬Ct)a Ratiob gene P E1 I1 E2 I2 E3 Probe A Probe B IL-1␣ 2.8 Ϯ 0.4 4.4 Ϯ 1.1 Ϫ1.6 IL-1␤ 25.7 Ϯ 2.3 146 Ϯ 37 Ϫ5.7 IL-2 1.8 Ϯ 0.4 1.7 Ϯ 0.7 ϩ1.1 IL-3 not detected not detected —– IL-4 4.4 Ϯ 1.4 0.42 Ϯ 0.8 ϩ10.5 NotI TaqI BamHI BamHI NotI IL-5 2.2 Ϯ 0.7 0.06 Ϯ 1.5 ϩ36.7 Targeting Neor E3E2I2 HSV-TK Vector IL-6 25.3 Ϯ 1.9 47.5 Ϯ 10.1 Ϫ1.9 Ϯ Ϯ ϩ SphI IL-7 1.2 0.2 0.5 1.0 2.4 IL-9 not detected not detected —– IL-10 13.5 Ϯ 8.8 2.9 Ϯ 1.0 ϩ4.7 IL-12␣ 3.4 Ϯ 0.6 12.8 Ϯ 5.2 Ϫ4.8 8.8 Kb IL-12␤ not detected not detected —– 4.4 Kb TGF-␤ 3.8 Ϯ 1.2 2.8 Ϯ 0.7 ϩ1.4 SacI BamHI TNF-␣ 17.7 Ϯ 6.3 25.7 Ϯ 7.1 Ϫ1.5 SphI NcoI TaqI BamHI SacI CCL1 KO IFN-␥ 25.0 Ϯ 7.1 100.5 Ϯ 64 Ϫ3.2 r gene Neo E3E2I2 CCL1 14.5 Ϯ 6.5 1.7 Ϯ 2.1 ϩ8.5 Downloaded from Probe A SphI Probe B CCL2 13.7 Ϯ 5.8 9.9 Ϯ 3.3 ϩ1.4 CCL3 51.3 Ϯ 8.2 53.5 Ϯ 19.2 Ϫ1.0 BC CCL4 33.0 Ϯ 9.5 79.0 Ϯ 5.7 Ϫ2.8 5’ Analysis 3’ Analysis b.p. CCL5 8.3 Ϯ 2.4 41.8 Ϯ 14 Ϫ5.0 -/--/- +/+ +/+ +/- +/- 390403 444 Kb 403 404 406 Kb 2036 CCL6 5.6 Ϯ 0.7 3.7 Ϯ 0.7 ϩ1.5 1636 Ϯ Ϯ ϩ 9 9 1018 CCL7 25.7 5.2 14.5 3.3 1.8 8 517 Ϯ Ϯ ϩ 7 8 KO allele CCL9/10 8.2 0.9 4.2 0.9 1.9 7 506 Ϯ Ϯ ϩ 6 http://www.jimmunol.org/ 6 396 CCL11 1.5 0.5 0.7 1.3 2.4 5 344 421 bp Ϯ Ϯ ϩ 4 5 CCL17 3.3 0.5 1.5 0.2 2.2 4 298 3 CCL20 3.7 Ϯ 0.3 4.1 Ϯ 0.6 Ϫ1.1 3 CCL22 3.4 Ϯ 1.8 2.3 Ϯ 1.0 ϩ1.5 2 CCL27 0.5 Ϯ 0.5 0.16 Ϯ 1.9 ϩ3.1 2 2036 1636 CXCL1 6.0 Ϯ 2.0 20.2 Ϯ 1.1 Ϫ3.4 1018 517 WT allele CXCL2 32.2 Ϯ 5.5 186 Ϯ 77.2 Ϫ5.8 1 506 Ϯ Ϯ Ϫ 1 396 604 bp CXCL5 33.5 2.5 274 80.8 8.2 344 CXCL9 14.0 Ϯ 2.0 19.6 Ϯ 1.6 Ϫ1.4 SphI Digestion SacI Digestion 298 CXCL10 28.8 Ϯ 7.2 65.2 Ϯ 4.9 Ϫ2.3 Probe A Probe B CXCL11 26.4 Ϯ 1.1 44.8 Ϯ 12.4 ϩ1.7 FIGURE 1.