Human T Cells That Are Able to Produce IL-17 Express the Chemokine Receptor CCR6 This information is current as Satya P. Singh, Hongwei H. Zhang, John F. Foley, Michael of September 26, 2021. N. Hedrick and Joshua M. Farber J Immunol 2008; 180:214-221; ; doi: 10.4049/jimmunol.180.1.214 http://www.jimmunol.org/content/180/1/214 Downloaded from References This article cites 58 articles, 27 of which you can access for free at: http://www.jimmunol.org/content/180/1/214.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 Human T Cells That Are Able to Produce IL-17 Express the Chemokine Receptor CCR61 Satya P. Singh, Hongwei H. Zhang, John F. Foley, Michael N. Hedrick, and Joshua M. Farber2 Some pathways of T cell differentiation are associated with characteristic patterns of chemokine receptor expression. A new -lineage of effector/memory CD4؉ T cells has been identified whose signature products are IL-17 cytokines and whose differen tiation requires the nuclear receptor, ROR␥t. These Th17 cells are critical effectors in mouse models of autoimmune disease. We have analyzed the association between chemokine receptor expression and IL-17 production for human T cells. Activating cord blood (naive) CD4؉ T cells under conditions driving Th17 differentiation led to preferential induction of CCR6, CCR9, and CXCR6. Despite these data, we found no strong correlation between the production of IL-17 and expression of CCR9 or CXCR6. ؉ By contrast, our analyses revealed that virtually all IL-17-producing CD4 T cells, either made in our in vitro cultures or found Downloaded from in peripheral blood, expressed CCR6, a receptor found on ϳ50% of CD4؉ memory PBL. Compared with CD4؉CD45RO؉CCR6؊ cells, CD4؉CD45RO؉CCR6؉ cells contained at least 100-fold more IL-17A mRNA and secreted 100-fold more IL-17 protein. The .CCR6؉ cells showed a similar enrichment in mRNA for ROR␥t. CCR6 was likewise expressed on all IL-17-producing CD8؉ PBL CCR6 has been associated with the trafficking of T, B, and dendritic cells to epithelial sites, but has not been linked to a specific T cell phenotype. Our data reveal a fundamental feature of IL-17-producing human T cells and a novel role for CCR6, suggesting both new directions for investigating IL-17-related immune responses and possible targets for preventing inflammatory http://www.jimmunol.org/ injury. The Journal of Immunology, 2008, 180: 214–221. ntigen activation drives naive T cells down any of a phenotypes and pathways of differentiation. For example, CXCR3 number of pathways of differentiation to yield highly is expressed preferentially on human Th1 cells (5–8), and its ex- A heterogeneous populations of effector/memory cells. pression is driven by T-bet (9, 10), the transcription factor that is Heterogeneity is a particularly prominent feature of the effector/ critical for Th1 differentiation (2). Conversely, CCR4 is expressed ϩ memory CD4 T cell population, which includes subsets capable preferentially on Th2 cells (5–8), and its expression can be driven of producing polarized patterns of cytokines that serve specialized by GATA-3 (9), the transcription factor that is critical for Th2 by guest on September 26, 2021 functions and have profound effects on the quality of the immune differentiation (2). It is likely that these receptors are components response (1). The varied effector/memory phenotypes are the con- of positive feedback loops to reinforce type 1 or type 2 responses sequences of changes in gene activities and related remodeling of in tissues, because CXCR3 ligands are induced by IFN-␥ (11), the chromatin, processes for which a number of transcription factors signature cytokine of Th1 cells, and CCR4 ligands are induced by have been identified as master regulators (2). Among the genes IL-4 (12), the signature cytokine of Th2 cells. induced as part of the programs of differentiation are those encod- Over the last several years, IL-17 and IL-17-producing effector/ ing adhesion molecules and chemoattractant receptors, which are memory T cells have been recognized as critical for producing responsible for the alterations in patterns of migration for effector/ tissue damage in mouse models of autoimmune disease, as well as memory vs naive cells (3, 4). in host defense against bacterial infection (reviewed in Ref. 13, The complexity of the T cell effector/memory population is re- 14). Much of the inflammatory damage previously ascribed to the flected in the expanded repertoire of chemokine receptors, from ϩ ϩ type 1 response is now understood to depend on IL-17 and on two or three species on naive CD4 or CD8 T cells, respectively, IL-23, the cytokine important for supporting the Th17 response in to at least fifteen species of chemokine receptors expressed in var- vivo (15, 16). Recently, it has been recognized that IL-17-produc- ious combinations on subsets of effector/memory cells. Although ϩ ing CD4 T cells (Th17 cells) form a lineage separate from Th1 the mechanisms underlying the regulation of chemokine receptor and Th2 cells, and that, in fact, the production of Th17 cells is expression on T cell subsets are poorly understood, some patterns inhibited by factors that support Th1/Th2 differentiation (17, 18). of receptor expression have been associated with well established Differentiation down the Th17 pathway depends on the orphan nuclear receptor ROR␥t (19), which likely serves as a master tran- Laboratory of Molecular Immunology, National Institute of Allergy and Infectious scriptional regulator for Th17 cells. Diseases, National Institutes of Health, Bethesda, MD 20892 There is a strong connection between the Th17 response and Received for publication April 24, 2007. Accepted for publication October 29, 2007. the chemokine system, because a major aspect of IL-17-driven 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 inflammation is neutrophil recruitment, for which the induction with 18 U.S.C. Section 1734 solely to indicate this fact. of CXC chemokines is an important component (13). An unan- 1 This research was supported by the Intramural Research Program of the National swered question, however, is whether or not there is a pattern of Institutes of Health, National Institute of Allergy and Infectious Diseases. chemokine receptor expression/chemokine responsiveness that 2 Address correspondence and reprint requests to Dr. Joshua M. Farber, Laboratory of is characteristic of IL-17-producing T cells. This question is Molecular Immunology, National Institute of Allergy and Infectious Diseases, Na- tional Institutes of Health, Building 10, Room 11N-111, 10 Center Drive, Bethesda, relevant for understanding the mechanisms of IL-17-induced MD 20892. E-mail address: [email protected] inflammation because IL-17 and/or IL-17-producing T cells are www.jimmunol.org The Journal of Immunology 215 Downloaded from FIGURE 1. Th17 differentiation of human cord blood CD4ϩ T cells in vitro. Human CD4ϩ T cells from cord blood were stimulated with immobilized plate bound anti-CD3 and anti-CD28 in nonpolarizing (NP), Th1, Th2, or Th17 conditions and harvested after 6 days. A, Cells cultured under conditions as noted above each dot plot were stimulated with the leukocyte activation mixture (see Materials and Methods) for 5 h, fixed, permeabilized, stained with http://www.jimmunol.org/ anti-IFN-␥-allophycocyanin and anti-IL-17A-Alexa Fluor 488, and analyzed by flow cytometry. The percentage of cells in each quadrant is noted. One representative experiment is shown of six performed. In this figure and in the other figures, separate experiments used cells from separate donors. B and C, Cells cultured under conditions as noted on the x-axes were stimulated with PMA and ionomycin for 4 h before isolating RNA. Levels of mRNAs for IL-17A (B) and ROR␥t(C) were quantified using real-time RT-PCR. Values were normalized to GAPDH and then to the results for the naive cells, which had been frozen at the time of isolation and thawed and treated with the activation mixture on day six. The data in B are means from three experiments and in C are from duplicate samples from one representative experiment of three performed. Error bars, SEM. found in the affected tissues and it is presumed that the cyto- The following cytokines and Abs were purchased from BD Pharmingen: by guest on September 26, 2021 kine’s presence at the site is critical for producing tissue dam- anti-IL-4, anti-IL-12, and anti-IFN-␥; anti-CCR3, anti-CCR4, anti- age (16, 20–23). It is possible, therefore, that blocking the entry CCR7, anti-CXCR1, anti-CXCR2, anti-CXCR3, anti-CXCR4, and anti-CXCR5, all conjugated to PE; anti-CCR5-PE-Cy5, anti- of Th17 cells into tissue – for example, by inhibiting one or CD4-allophycocyanin-Cy7, anti-CD8-FITC, anti-CD45RO-PECy5, more chemokine receptors – might be an effective way of ame- anti-IFN-␥-FITC and anti-IFN-␥-allophycocyanin.