CCR 7 Ligands Are Required for Development of Experimental Autoimmune Encephalomyelitis through Generating IL-23-Dependent Th17 Cells This information is current as of September 24, 2021. Taku Kuwabara, Fumio Ishikawa, Takuwa Yasuda, Kentaro Aritomi, Hideki Nakano, Yuriko Tanaka, Yayoi Okada, Martin Lipp and Terutaka Kakiuchi J Immunol 2009; 183:2513-2521; Prepublished online 22 July 2009; Downloaded from doi: 10.4049/jimmunol.0800729 http://www.jimmunol.org/content/183/4/2513 http://www.jimmunol.org/ References This article cites 40 articles, 15 of which you can access for free at: http://www.jimmunol.org/content/183/4/2513.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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The Journal of Immunology CCR 7 Ligands Are Required for Development of Experimental Autoimmune Encephalomyelitis through Generating IL-23-Dependent Th17 Cells1 Taku Kuwabara,* Fumio Ishikawa,* Takuwa Yasuda,2* Kentaro Aritomi,*‡ Hideki Nakano,3* Yuriko Tanaka,* Yayoi Okada,* Martin Lipp,§ and Terutaka Kakiuchi4*† CCL19 and CCL21 are thought to be critical for experimental autoimmune encephalomyelitis (EAE) induction, but their precise role is unknown. We examined the role of these chemokines in inducing EAE. C57BL/6 mice lacking expression of these chemokines (plt/plt mice) or their receptor CCR7 were resistant to EAE induced with myelin oligodendrocyte glyco- protein peptide 35–55 (MOG35–55) and pertussis toxin. However, passive transfer of pathogenic T cells from wild-type mice induced EAE in plt/plt mice, suggesting a defect independent of the role of CCR7 ligands in the migration of immune cells. Downloaded from Examination of draining lymph node (DLN) cells from MOG35–55-immunized plt/plt mice found decreased IL-23 and IL-12 production by plt/plt dendritic cells (DCs) and a concomitant defect in Th17 cell and Th1 cell generation. In contrast, production of the Th17 lineage commitment factors IL-6 and TGF-␤ were unaffected by loss of CCR7 ligands. The adoptive transfer of in vitro-generated Th17 cells from DLN cells of MOG35–55-immunized plt/plt mice developed EAE in wild-type recipient mice, whereas that of Th1 cells did not. Pathogenic Th17 cell generation was restored in plt/plt DLNs with the addition of exogenous IL-23 or CCL19/CCL21 and could be reversed by inclusion of anti-IL-23 mAb in cultures. Exogenous http://www.jimmunol.org/ CCL19/CCL21 induced IL-23p19 expression and IL-23 production by plt/plt or wild-type DCs. Therefore, CCR7 ligands have a novel function in stimulating DCs to produce IL-23 and are important in the IL-23-dependent generation of pathogenic Th17 cells in EAE induction. The Journal of Immunology, 2009, 183: 2513–2521. xperimental autoimmune encephalomyelitis (EAE)5 is an reactivation upon encountering endogenous Ag leading to nerve autoimmune disease of the CNS in mouse and rat that demyelination. Th17 cells, a helper CD4ϩ T cell lineage generated E serves as a disease model for human multiple sclerosis by IL-6, TGF-␤, and IL-23, are the pathogenic cells in EAE and (1). EAE is induced through sensitization with neuroantigens such are capable of inducing EAE in recipient mice by passive transfer by guest on September 24, 2021 as myelin oligodendrocyte glycoprotein (MOG) that activates neu- (2). The activation and activity of pathogenic T cells appear de- roantigen-reactive T cells in the peripheral lymphoid organs. Sub- pendent on the coordinated migration of several cell types, a phe- sequent migration of these T cells into the CNS leads to their nomena regulated by chemokines (3). Indeed, many chemokines have been shown to be critical for the development of EAE, mainly in the context of recruitment of immune cells into the CNS (4). *Department of Immunology and †Advanced Medical Research Center, Toho Uni- versity School of Medicine, Tokyo, Japan; ‡Department of Orthopedic Surgery, Jun- Chemokines may regulate induction of pathogenic T cells inde- tendo University School of Medicine, Tokyo, Japan; and §Department of Molecular pendent of their role in migration of immune cells but their role in Tumor Genetics and Immunogenetics, Max-Delbruck Center for Molecular Medicine, inducing pathogenic T cells in EAE is unclear. Berlin, Germany Using mutant mice lacking the expression of CCL19 and Received for publication March 6, 2008. Accepted for publication June 21, 2009. CCL21 ( plt/plt mice), we previously showed that these CC che- 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 mokine ligands, which share the CCR7, not only play a critical role with 18 U.S.C. Section 1734 solely to indicate this fact. in the migration of naive T cells and dendritic cells (DCs) into 1 This work was supported in part by Project Research of Toho University School of secondary lymphoid organs, but also regulate in vivo T cell re- Medicine (to T.Ku., F.I., and Y.O.), the Research Promotion Grants from Toho Uni- sponses to an Ag (5–8). Based on these findings, we hypothesized versity Graduate School of Medicine (Grant 05-02 to T.Ka., Grant 07-02 to T.Ku., and Grant 08-02 to Y.T.), and Grants-in-Aid for Scientific Research from the Japan that these chemokines regulate not only the migration of patho- Society for the Promotion of Science to T.Ka. (Grants 17590900 and 19591013), to genic T cells into the CNS, which was suggested previously (9), T.Ku. (Grant 18790605), and to Y.K. (Grant 19790695). but also the induction of pathogenic T cells. In the present study, 2 Current address: Laboratory for Immunogenetics, RIKEN Research Center for Al- we examined the role of the CCL19 and CCL21 chemokines in the lergy and Immunology, Yokohama 230-0045, Japan. induction of EAE using plt/plt mice. Deficiency in the expression 3 Current address: Laboratory of Respiratory Biology, National Institute of Environ- mental Health Sciences. National Institutes of Health, 111 T.W. Alexander Drive, of these chemokines was protective against development of EAE Building 101, E244, Research Triangle Park, NC 27709. due to a defect in IL-23-dependent induction of Th17 cells; how- 4 Address correspondence and reprint requests to Dr. Terutaka Kakiuchi, Department ever, passively transferred pathogenic T cells could initiate EAE in of Immunology, Toho University School of Medicine, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143-8540, Japan. E-mail address: [email protected] plt/plt mice. Thus, chemokines CCL19 and CCL21 are crucial for the induction of pathogenic T cells, independent of their role in 5 Abbreviations used in this paper: EAE, experimental autoimmune encephalomyeli- tis; DC, dendritic cell; BMDC, bone marrow-derived DC; LN, lymph node; DLN, immune cell migration. In this study, we provide the first descrip- draining LN; MOG, myelin oligodendrocyte glycoprotein; rm, recombinant mouse; tion of a novel function of these CCR7 ligands, namely, the stim- WT, wild type; PT, pertussis toxin. ulation of DCs to produce IL-23, resulting in the generation of Copyright © 2009 by The American Association of Immunologists, Inc. 0022-1767/09/$2.00 Th17 cells. www.jimmunol.org/cgi/doi/10.4049/jimmunol.0800729 2514 ROLE OF CCL19/21 IN IL-23 PRODUCTION AND EAE DEVELOPMENT Materials and Methods real-time PCR was conducted using TaqMan Gene Expression Assay Kits Mice (Applied Biosystems), Mm00434165_m1 for IL-12 and Mm00518984_m1 for IL-23 on an Applied Biosystems Prism 7000 Sequence Detector Sys- C57BL/6 mice were obtained from Charles River Laboratories Japan and tem. GAPDH was used as an endogenous reference for normalization. C57BL/6-plt/plt and C57BL/6-CCR7Ϫ/Ϫ mice were bred and kept at the Quantitative real-time PCR experiments were repeated twice in triplicate. Toho University School of Medicine animal facility (7) under specific To analyze the expression of IL-23p19 mRNA and IL-12p35 mRNA in ϩ pathogen-free conditions in accordance with the institutional guidelines. DLN DCs, CD11c cells were enriched from DLN cells using a magnetic All animal experiments were approved by the institutional review board. positive selection kit (BD Biosciences). Mice were used at 8–12 wk of age. Where indicated, CD45.1ϩ C57BL/ 6-Ly5.1 mice were also used. These mice were maintained in our animal Bone marrow-derived DCs (BMDCs) facility. The peripheral lymph node (LN) was much smaller in plt/plt mice than in wild-type (WT) mice (5). The mean cell number in an experiment BMDCs were prepared from bone marrow of naive C57BL/6 mice as pre- ϫ 6 was 4.2 ϫ 105 cells/LN in the inguinal and axillary LNs from six naive viously described (10). Where indicated, 4 10 cells in 10 ml of RPMI plt/plt mice, whereas 1.5 ϫ 106 cells/LN in naive WT mice. Nine days after 1640 containing rmGM-CSF (PeproTech/Tebu) were pulsed with rm- EAE induction as described below, the mean cell numbers were 6.7 ϫ 106 CCL19 or rmCCL21 (R&D Systems).