Epithelial Inflammation Is Associated with CCL28 Production and the Recruitment of Regulatory T Cells Expressing CCR10 This information is current as Bertus Eksteen, Alice Miles, Stuart M. Curbishley, Chris of September 26, 2021. Tselepis, Allister J. Grant, Lucy S. K. Walker and David H. Adams J Immunol 2006; 177:593-603; ; doi: 10.4049/jimmunol.177.1.593 http://www.jimmunol.org/content/177/1/593 Downloaded from References This article cites 51 articles, 22 of which you can access for free at: http://www.jimmunol.org/content/177/1/593.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 © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Epithelial Inflammation Is Associated with CCL28 Production and the Recruitment of Regulatory T Cells Expressing CCR101 Bertus Eksteen,* Alice Miles,* Stuart M. Curbishley,* Chris Tselepis,‡ Allister J. Grant,* Lucy S. K. Walker,† and David H. Adams2* Mucosal tissues require constant immune surveillance to clear harmful pathogens while maintaining tolerance to self Ags. Reg- ␣ ␤ ulatory T cells (Tregs) play a central role in this process and expression of E 7 has been reported to define a subset of Tregs with tropism for inflamed tissues. However, the signals responsible for recruiting Tregs to epithelial surfaces are poorly understood. We have isolated a subset of CCR10-expressing CD25؉CD4؉Foxp3؉ Tregs with potent anti-inflammatory properties from chronically inflamed human liver. The CCR10؉ Tregs were detected around bile ducts that expressed increased levels of the CCR10 ligand CCL28. CCL28 was secreted by primary human cholangiocytes in vitro in response to LPS, IL-1␤, or bile acids. Exposure of ؉ CCR10 Tregs to CCL28 in vitro stimulated migration and adhesion to mucosal addressin cell adhesion molecule-1 and VCAM-1. Downloaded from Liver-derived CCR10؉ Tregs expressed low levels of CCR7 but high levels of CXCR3, a chemokine receptor associated with ؉ ␤ ␣ infiltration into inflamed tissue and contained a subset of E 7 cells. We propose that CXCR3 promotes the recruitment of Tregs to inflamed tissues and CCR10 allows them to respond to CCL28 secreted by epithelial cells resulting in the accumulation of .CCR10؉ Tregs at mucosal surfaces. The Journal of Immunology, 2006, 177: 593–603 hronic inflammation leads to tissue damage and the re- important determinant of Treg function because CTLA-4-deficient http://www.jimmunol.org/ lease of multiple potential autoantigens. Although sec- mice have a similar phenotype to Foxp3 deficiency (8, 7). A recent C ondary immune responses against such Ags can be de- study suggests that Tregs do not express the inhibitory receptor tected, they are not a dominant feature of most chronic programmed cell death-1 (PD-1) on their surface, although it is inflammatory diseases suggesting that mechanisms exist to sup- retained intracellularly. This finding discriminates them from press the adaptive immune response in inflamed tissues. Regula- CD4ϩ/CD25ϩ effector cells that express high levels of cell surface tory T cells (Tregs)3 have evolved to limit the local damage re- PD-1 (9). sulting from infectious challenges to the host. Natural Tregs arise Tregs control inflammation by contact-dependent TGF-␤ and in the thymus and survive as well as operate in the periphery by IL-10 production (10, 11) and are able to control experimental gut by guest on September 26, 2021 responding to a large variety of self-Ags (1–3). Tregs are CD4 inflammation in adoptive transfer models (12). Paradoxically, cells that have a distinct phenotype characterized by expression of Tregs are required to maintain chronic intestinal inflammation in the a subunit of the high affinity IL-2R, CD25, and the glucocor- animal models, presumably by dampening more aggressive acute ticoid-induced TNFR (4). Despite displaying diverse TCR, there is inflammation (13). Epithelial surfaces in particular are vulnerable evidence to suggest Tregs have a higher propensity to recognize Ϫ to invasion by microbes and are a frequent target of chronic in- self peptides than conventional CD25 T cells (3, 5). The most flammatory diseases (14). Although Tregs have been reported in specific Treg marker is the transcription factor Foxp3, which is inflamed peripheral tissues, little is known about their function or critical for Treg function. Retroviral transfer of Foxp3 to naive T the homing mechanisms that localize them to epithelial sites (15). cells converts them into functional Tregs, whereas its deletion ab- The chemokine receptor CCR10 is detected on both T and B lates regulatory function and triggers autoimmunity (6, 7). Tregs lymphocytes at epithelial sites (16, 17) and defines subsets of lym- also constitutively express the negative regulatory receptor phocytes that can be recruited to either mucosal or cutaneous ep- CTLA-4, which binds the ligands CD80/CD86 and may be an ithelial sites. Mucosal homing is driven by the ligand CCL28, which is expressed by columnar epithelia in the gut, lung, breast *Liver Research Laboratories, †Medical Research Council Centre for Immune Reg- and salivary glands (18), whereas homing to the skin is triggered ulation, Institute for Biomedical Research, and ‡Epithelial Research Group, Univer- by the alternative CCR10 ligand, CCL27 (19). Specificity is en- sity of Birmingham, Birmingham, United Kingdom hanced by the coexpression of CCR10 with organ-specific adhe- Received for publication October 14, 2005. Accepted for publication April 12, 2006. sion receptors. Thus mucosal CCR10ϩ lymphocytes also express The costs of publication of this article were defrayed in part by the payment of page ␣ ␤ required for recruitment to the gut, whereas CCR10ϩ lym- charges. This article must therefore be hereby marked advertisement in accordance 4 7 with 18 U.S.C. Section 1734 solely to indicate this fact. phocytes, which show tropism for the skin, coexpress the skin- 1 This work was supported by grants from The Medical Research Council and Core homing receptor CCR4 and the cutaneous lymphocyte Ag (19). Charity, U.K. CCL28 is constitutively expressed in the colon and increased by 2 Address correspondence and reprint requests to Dr. David H. Adams, Liver Re- proinflammatory cytokines (20) and bacterial products, suggesting search Group, Medical Research Council Centre for Immune Regulation, 5th Floor it has a role in recruiting effector cells to areas of epithelial injury Institute for Biomedical Research, University of Birmingham, Birmingham B15 2TH, U.K. E-mail address: [email protected] (21). CCR10 expression has also been reported on nonlymphoid 3 Abbreviations used in this paper: Treg, regulatory T cell; LIL, liver-infiltrating malignant cells (22), and coexpression of CCR9 and CCR10 has lymphocyte; PSC, primary sclerosing cholangitis; PBC, primary biliary cirrhosis; been implicated in the formation of small bowel melanoma me- ALD, alcoholic liver disease; MAdCAM-1, mucosal addressin cell adhesion mole- ϩ cule-1; hpf, high-power field; BEC, biliary epithelial cell; CDCA, chenodeoxycholic tastases (23). CCR10 lymphocytes are positioned in the intraepi- ␣ ␤ acid. thelial compartment where coexpression of E 7 integrins (24) Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 594 CCR10 EXPRESSION ON EPITHELIAL Tregs allows them to interact with E-cadherin expressed at epithelial ad- phocytes were separated using 33%/77% (v/v) Percoll (Amersham Bio- ␣ sciences) density gradient centrifugation at 2000 rpm (650 ϫ g) for 30 min. herens junctions (25). The expression of E integrins also defines ϩ a population of CD25 Tregs with enhanced suppressive proper- PBL isolation ␣ Ϫ ϩ ties compared with E CD25 Tregs (26). Recent studies have suggested that the function of ␣ ϩ Tregs is at least in part depen- Lymphocytes were isolated from peripheral venous blood and diluted 1/1 E with PBS before centrifuged over a Lymphoprep (Invitrogen Life Tech- dent on their ability to be recruited to inflamed tissue compart- nologies) gradient for 30 min at 2000 rpm (650 ϫ g). ments (27). Experimental animals that lack fucosultransferase re- quired to synthesize E- and P-selectin ligands during inflammation Isolation and culture of BEC ␣ ϩ are unable to recruit E effector/memory Tregs to inflammatory BEC were isolated according to previously described methods (30). sites and as a consequence display a reduced ability to suppress Briefly, liver tissue was finely chopped and subjected to enzyme digestion ϩ␣ Ϫ (collagenase type IV) and density gradient centrifugation. Nonparenchymal inflammation (27). In contrast, naive Tregs (CD25 E ) express CCR7 and CD62L, allowing them to enter lymphoid tissues to cells were then removed and separated by immunomagnetic selection. Cells positive for mAb