CCR4 Involvement in the Expansion of T Helper Type 17 Cells in a Mouse Model of Psoriasis

Psoriasis is a chronic skin disease associated with T helper (Th)17-mediated inflammation. Because CCR4 major chemokine receptor expressed on Th17 cells, we investigated the role of in modified imiquimod-induced psoriasis model that showed enhanced infiltration cells. CCR4-deficient mice had less severe than wild-type mice. cells were decreased lesions and regional lymph nodes In mice, CD44+ memory expressing found to be clustered dendritic CCL22, ligand for CCR4. Such cell?Th17 cell clusters significantly Similar results obtained using IL-23?induced model. vitro, compound 22, antagonist, reduced expansion coculture CD11c+ CD4+ separately prepared from psoriasis. vivo, 22 ameliorated psoriasis-like significant decreases lesions. Collectively, likely play pathogenesis through an autoimmune characterized by erythematous plaques covered silvery scales, epidermal hyperplasia, dermal immune (Boehncke, 2015Boehncke W.H. Etiology psoriasis.Rheum Dis Clin North Am. 2015; 41: 665-675Abstract Full Text PDF PubMed Scopus (108) Google Scholar). identification IL-17?producing (Th)17 as novel subset effector numerous studies have shown are highly autopathogenic can induce diseases, including (Brembilla et al., 2018Brembilla N.C. Senra L. Boehncke The IL-17 family cytokines psoriasis: IL-17A beyond.Front Immunol. 2018; 9: 1682Crossref (190) Scholar; Dardalhon 2008Dardalhon V. Korn T. Kuchroo V.K. Anderson A.C. Role Th1 organ-specific autoimmunity.J Autoimmun. 2008; 31: 252-256Crossref (315) Di Cesare 2009Di A. Meglio P. Nestle F.O. IL-23/Th17 axis immunopathogenesis psoriasis.J Invest Dermatol. 2009; 129: 1339-1350Abstract (809) IL-17A, member cytokine family, which mainly secreted induces proliferation aberrant differentiation keratinocytes also promotes expression various proinflammatory and/or chemokines keratinocytes, fibroblasts Chemokines large small, structurally related orchestrate migration tissue localization corresponding receptors (Zlotnik Yoshie, 2012Zlotnik Yoshie O. superfamily revisited.Immunity. 2012; 36: 705-716Abstract (693) Mature now known composed functional subsets, each unique set For example, Th2 regulatory been express CCR4, TARC/CCL17 MDC/CCL22 (Imai 1999Imai Nagira M. Takagi S. Kakizaki Nishimura Wang J. al.Selective recruitment CCR4-bearing toward antigen-presenting CC thymus activation-regulated macrophage-derived chemokine.Int 1999; 11: 81-88Crossref (598) Matsushima, 2015Yoshie Matsushima K. its ligands: bench bedside.Int 27: 11-20Crossref (205) Zlotnik particular, coexpress CCR6 (Acosta-Rodriguez 2007Acosta-Rodriguez E.V. Rivino Geginat Jarrossay D. Gattorno Lanzavecchia al.Surface phenotype antigenic specificity human interleukin 17-producing cells.Nat 2007; 8: 639-646Crossref (1413) Previously, Hedrick 2009Hedrick M.N. Lonsdorf A.S. Shirakawa A.K. Richard Lee C.C. Liao F. Singh S.P. al.CCR6 required IL-23-induced inflammation mice.J Invest. 119: 2317-2329Crossref (181) Scholar IL-23?induced, IL-22?mediated mouse (Hedrick They CCR6-deficient protected development psoriatic disease. Unexpectedly, however, no difference skin-infiltrating (WT) Subsequently, model, Mabuchi al. demonstrated was involved IL-22?producing ?? instead (Mabuchi 2013Mabuchi T.P. Takekoshi Jia G.F. Wu X. Kao M.C. trafficking ??-T psoriasiform dermatitis.J 2013; 133: 164-171Abstract (101) Similarly, imiquimod (IMQ)-induced Cochez 2017Cochez P.M. Michiels C. Hendrickx E. Dauguet N. Warnier G. Renauld J.C. al.Ccr6 dispensable induced despite effect homing IL-22-producing cells.J 2017; 137: 1094-1103Abstract (13) Thus, Th17-mediated remains obscure. There even information However, our previous study has plays substantial another Th17-associated inflammatory CNS, is, experimental encephalomyelitis (Moriguchi 2016Moriguchi Miyamoto Tanaka Ueno R. Nakayama al.C-C type 4 antagonist ameliorates encephalomyelitis.J Neuroimmunol. 2016; 291: 54-58Abstract (11) Scholar), although precise discovered. On activation antigen loading, (DCs) localized migrate (LNs). Migrated DCs attract cognate receptors. This cluster formation subsequent antigen-specific (Lian Luster, 2015Lian Luster A.D. Chemokine-guided positioning node orchestrates generation adaptive responses.Curr Opin Cell Biol. 1-6Crossref (43) Worbs 2017Worbs Hammerschmidt S.I. Förster Dendritic health disease.Nat Rev 17: 30-48Crossref (380) Katou 2001Katou Ohtani H. Ono Saaristo al.Macrophage-derived (MDC/CCL22) lymphocyte-dendritic inflamed secondary lymphoid tissue.Am J Pathol. 2001; 158: 1263-1270Abstract (86) observed involving CCR4-expressing CCL22-expressing chronically tissues LNs. they unable identify such clusters. Recently, CCL22 form LNs bearing B16 melanoma skin. DC?Th17 crucial tumor-bearing (Matsuo 2016Matsuo Itoh Koyama Imamura Kawai Nishiwaki al.CCR4 critically effective antitumor immunity intradermal melanoma.Cancer Lett. 378: 16-22Crossref (19) Furthermore, induction CD8+ T-cell responses against Others reported promote diseases (Ankathatti Munegowda 2011Ankathatti Deng Y. Chibbar Xu Q. Freywald Mulligan S.J. al.A distinct Th17- Th17-stimulated CTL 1 diabetes 2011; 811-826Crossref (20) Martin-Orozco 2009Martin-Orozco Muranski Chung Yang X.O. Yamazaki Lu al.T 17 cytotoxic tumor immunity.Immunity. 787-798Abstract (574) CCL22–CCR4 may promotion interactions. Therefore, this study, models psoriasis, (Di Topical application IMQ 5?6 days used trigger (van der Fits 2009van Mourits Voerman J.S. Kant Boon Laman J.D. al.Imiquimod-induced mediated via IL-23/IL-17 axis.J 182: 5836-5845Crossref (1251) unlike represent only minor fraction lesion, skin-homing source (Cochez Van Belle 2012Van A.B. de Heusch Lemaire M.M. Dunussi-Joannopoulos al.IL-22 188: 462-469Crossref (224) van initial experiments, standard protocol differences between WT terms severity (ear thickness, erythema, scaling) or presence (Supplementary Figure S1a b). longer period sensitization might necessary performed extended every other day 14 days. We compared short-term long-term (Figure 1a). Skin visible 2?4 increased up end both 1b). Although appeared histologically macroscopically similar 1c), tended ear scaling, cumulative score treatment 1b d). next examined (CD4+IL-17+ cells), (??TCR+IL-17+ neutrophils (CD11b+Ly-6G+ macrophages (CD11b+F4/80+ conventional type-1 (CD103+CD11c+MHC II+ type-2 (CD11b+CD11c+MHC cells) As 1e, LNs, those protocol. not statistically significant, hand, there two models. These suggested more dominant findings mentioned earlier, therefore decided use investigate responses. 2a, progression clearly after 6 H&E staining much hyperplasia 2b c). apparent 6-day lag deficiency interesting due time enumerated 14. fewer but 2d). confirmed infiltrating positive CCR6, whereas 2e). quantitated mRNA levels Th17-related (Il-17a, Il-22, Il-23) 3a, these their protein 3c). psoriasis-related genes, signal transducer activator transcription gene Stat3 keratin K14, level 3 phosphorylation elevated 3a (Diani 2016Diani Altomare Reali Helper subsets clinical manifestations Immunol Res. 2016: 7692024Crossref (72) T-cell?related namely, Ifn-? granzyme B Gzmb. genes 3a). Consistently, flow cytometry analysis 3d). (Bachelerie 2013Bachelerie Ben-Baruch Burkhardt A.M. Combadiere Farber J.M. Graham G.J. al.International Union Basic Clinical Pharmacology. [corrected]. LXXXIX. Update introducing new nomenclature atypical [published correction appears Pharmacol 2014;66:467].Pharmacol Rev. 66: 1-79Crossref (546) Il4 Foxp3. Their Flow increases (Chan 2006Chan J.R. Blumenschein W. Murphy Diveu Wiekowski Abbondanzo al.IL-23 stimulates TNF IL-20R2-dependent mechanisms implications pathogenesis.J Exp Med. 2006; 203: 2577-2587Crossref (539) Recombinant IL-23 intradermally injected into 16 4a, all erythema By staining, 4b ??T 4d). patients CCL20, CCL17 ligands strongly (Homey 2000Homey B. Dieu-Nosjean Wiesenborn Massacrier Pin J.J. Oldham al.Up-regulation macrophage protein-3 alpha/CCL20 2000; 164: 6621-6632Crossref (451) Leung 2004Leung D.Y. Boguniewicz Howell M.D. Nomura I. Hamid Q.A. New insights atopic 2004; 113: 651-657Crossref (1166) IMQ-induced CCL20 5a). cytometry, isolated marker CD44 5b). immunohistochemistry, further produced small round 5c). double CCL22-producing IL-17A?producing counting LN sections 14, numbers 5d). CCR4–CCL22 expand To examine involvement vitro experiment. IMQ-sensitized cocultured without (Purandare 2007Purandare A.V. Wan Somerville J.E. Burke Vaccaro al.Core exploration optimization antagonists.Bioorg Med Chem 679-682Crossref (42) 5e, expanded DCs, poor when 5f). recombinant efficiently attracted chemotaxis assay 5g). supported promoting interactions so far could therapeutic target test hypothesis, tested vivo. 6a, scaling dose-dependent manner. revealed treated 6b 6d). 6e). namely Il-17a, Il-23, activated Stat3, 6f). one most employing (5?7 days) topical weakly 1), skin-resident extending 1). current Using contributes DC–Th17 5). Of note, evident 2). probably length Scholar) 4). nature often comorbid bullous vitiligo, thyroiditis enforced discovery autoantigens, cathelicidin ADAMTSL5, autoreactive self-antigens addition context, autoimmunity supports cell?mediated CCR10 roles blood regard, CCR6-expressing lesion (Guttman-Yassky 2007Guttman-Yassky Lowes M.A. Fuentes-Duculan Whynot Novitskaya Cardinale al.Major products distinguish dermatitis Allergy 1210-1217Abstract (174) 2012Mabuchi Chang T.W. Quinter Hwang S.T. Chemokine therapy Dermatol Sci. 65: 4-11Abstract (73) CCL27, CCR10, predominantly normal skin, (Riis 2011Riis J.L. Johansen Vestergaard Bech Kragballe Iversen Kinetics differential skin-related CCL27 allergic contact dermatitis.Exp 20: 789-794Crossref 5a), (data shown). main cause Indeed, little impact number (Leung it recently serum correlates at least some (Mehta 2013Mehta N.N. Li Szapary Krueger Brodmerkel Modulation cardiometabolic pathways Transl 194Crossref (35) 2019Xu Zhu Han Yan al.In-depth proteomics reveals biomarkers response traditional Chinese medicine.Theranostics. 2019; 2475-2488Crossref (39) transdermal delivery resulting augmented scores S2). inducing angiogenic factor VEGF (Heidenreich 2009Heidenreich Röcken Ghoreschi Angiogenesis drives pathogenesis.Int 90: 232-248Crossref (244) contribute exacerbation erythema. Previous IL-22, potential targets drive maintain Accordingly, anti?IL-17A mAb ixekizumab anti?IL-12/23 ustekinumab approved 6). Given anti-CCR4 mogamulizumab refractory adult lymphoma leukemia cutaneous lymphomas (Yoshie molecule blockers useful either alone combination anti?IL-23 mAb. conclusion, important possibly thus diseases. See Supplementary Materials Methods methods. C57BL/6J background purchased Jackson Laboratory (Bar Harbor, ME) maintained >20 generations. CLEA Japan (Tokyo, Japan). Mice specific pathogen-free conditions ages 6?10 weeks. All animal experiments Center Animal Experiments, Kindai University, Higashiosaka, Japan, accordance institutional guidelines. 5% cream (Beselna cream, Mochida Pharmaceutical, Tokyo, Japan) applied auricle (0.8 mg per ear) (the model) present model). 20 ?l (Abcam, Cambridge, MA) PBS (25 ?g/ml) Disease activity evaluated PASI. Erythema, thickness scored scale 0 4—0 indicating none, slight, 2 moderate, marked, very marked (erythema + thickness) served measure 12. Ear measured dial gauge (OZAKI MFG, selective intraperitoneal injection 0.1 0.5 ?g day. synthesized 99.5% purity basis published (Yamamoto 2018Yamamoto Matsuo Nagakubo Higashiyama Oiso enhances DC shows potent vaccine adjuvant inhibition recruitment.J 136: 165-171Crossref (8) Briefly, sorted biotinylated anti–mouse CD11c antibody (clone N418) Streptavidin Particles Plus (BD Biosciences, San Jose, CA) Separation Magnet Biosciences). Mouse CD4 Lymphocyte Enrichment Set (105 (106 percentages CD4+IL-17A+ analyzed cytometry. Student's t-test analyze groups. One-way ANOVA Holm post hoc multiple considered P < 0.05 significant. No datasets generated during study. Kazuhiko Matsuo: https://orcid.org/0000-0001-5782-5300 Kosuke Kitahata: https://orcid.org/0000-0002-4120-1859 Yuichiro Kaibori: https://orcid.org/0000-0001-5323-2647 Yuka Arima: https://orcid.org/0000-0003-1439-6563 Arisa Iwama: https://orcid.org/0000-0002-2469-0670 Mana Ito: https://orcid.org/0000-0001-8616-1718 Yuta Hara: https://orcid.org/0000-0002-7389-6480 Daisuke Nagakubo: https://orcid.org/0000-0003-1213-1246 Ying-Shu Quan: https://orcid.org/0000-0002-2430-8507 Fumio Kamiyama: https://orcid.org/0000-0002-6831-2807 Naoki Oiso: https://orcid.org/0000-0001-5593-6496 Akira Kawada: https://orcid.org/0000-0002-1868-2295 Osamu Yoshie: https://orcid.org/0000-0003-4353-5809 Takashi Nakayama: https://orcid.org/0000-0002-8493-899X authors state conflict interest thank Takanori Naganuma, Junko Takahara, Rio Nishikawa excellent technical assistance. work Ministry Education, Culture, Sports, Science Technology –Supported Program Strategic Research Foundation Private Universities, 2014–2018 (S1411037 TN) University Fund Antiaging Project (TN). Division Joint Center, LSRForttessa FACSAria. Conceptualization: KM, KK, TN; Data Curation: KK; Formal Analysis: Funding Acquisition: NO, AK, Investigation: YK, YA, AI, MI, YH, DN; Methodology: YH; Administration: Resources: YSQ, FK; Supervision: OY, Validation: Writing - Original Draft Preparation: – Review Editing: TN Download .pdf (.26 MB) Help pdf files