A Critical Role for Thymic Stromal Lymphopoietin in Nickel-Induced Allergy in Mice This information is current as Meinar Nur Ashrin, Rieko Arakaki, Akiko Yamada, of September 28, 2021. Tomoyuki Kondo, Mie Kurosawa, Yasusei Kudo, Megumi Watanabe, Tetsuo Ichikawa, Yoshio Hayashi and Naozumi Ishimaru J Immunol 2014; 192:4025-4031; Prepublished online 26 March 2014; Downloaded from doi: 10.4049/jimmunol.1300276 http://www.jimmunol.org/content/192/9/4025 Supplementary http://www.jimmunol.org/content/suppl/2014/03/26/jimmunol.130027 http://www.jimmunol.org/ Material 6.DCSupplemental References This article cites 37 articles, 7 of which you can access for free at: http://www.jimmunol.org/content/192/9/4025.full#ref-list-1 Why The JI? Submit online. by guest on September 28, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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The Journal of Immunology A Critical Role for Thymic Stromal Lymphopoietin in Nickel-Induced Allergy in Mice Meinar Nur Ashrin,*,† Rieko Arakaki,* Akiko Yamada,* Tomoyuki Kondo,* Mie Kurosawa,* Yasusei Kudo,* Megumi Watanabe,† Tetsuo Ichikawa,† Yoshio Hayashi,* and Naozumi Ishimaru* Ni is the most frequent cause of contact allergy induced by metals. However, the underlying mechanism of this induction is unknown. Our previous research demonstrates that activation of dendritic cells (DCs) through p38MAPK/MKK6 is required for Ni-induced allergy in mice. In the current study, we investigated the cellular and molecular mechanisms underlying Ni-induced allergy using a mouse model that involves injecting Ni into the ear, with or without Freund’s incomplete or complete adjuvants. Nickel had greater potential to cause allergic reactions compared with palladium and gold. Among the proteins expressed at higher levels in mice with Ni-induced allergy, we focused on thymic stromal lymphopoietin (TSLP), which is produced in abundance by keratinocytes. We Downloaded from detected increased expression of the TSLP receptor (TSLPR) in DCs from cervical lymph nodes of mice with Ni-induced allergy, suggesting that DCs in ear tissues were activated through TSLPR signaling induced by keratinocyte-derived TSLP. Furthermore, delayed-type hypersensitivity reactions in mice with Ni-induced allergy were decreased significantly by injection of a Tslp–short interfering RNA along with atelocollagen in the ear skin. These results suggest that Ni allergy may be triggered by a TSLP/TSLPR- mediated interaction between epithelial and immune cells. The Journal of Immunology, 2014, 192: 4025–4031. http://www.jimmunol.org/ e continuously encounter materials containing Au, Ag, as Langerhans cells or dendritic cells (DCs), are induced by certain Hg, Ni, Ti, Cr, and Co in our daily lives. For example, cytokines, including IL-1b and TNF-a, which are produced by W Ni-based alloys are widely used in costume jewelry epithelial cells (16–19). APCs migrate to draining lymph nodes and dental materials, and Ni frequently causes contact allergy where they present allergens or haptens to naive T cells. Subse- categorized as type IV delayed-type hypersensitivity (DTH), with quent re-exposure to the same allergen or hapten leads to a hyper- skin inflammation mediated by hapten-specific T cells (1–5). sensitivity reaction during the effector phase (19). However, the Approximately 20% of women and 6% of men are sensitized to Ni precise molecular mechanisms that mediate interactions between (6). Symptoms of Ni allergy appear in the oral mucosa and are epithelial and immune cells in Ni allergy are unknown. by guest on September 28, 2021 widely observed in the skin (7–9). We reported previously that Ni-activated dermal DCs play an Ni ions (Ni2+) released from various alloys are potent allergens important role in the development of allergic reactions through the or haptens that can trigger inflammation of the skin (10–12). Ni2+ activation of p38 MAPK in a model system in which mice are penetrates the skin and activates epithelial cells that produce cyto- exposed to Ni combined with adjuvants (20). Although Langerhans kines or chemokines following hyperreactive immune responses cells or dermal DCs play fundamental roles in the initial phase of involving the activation of APCs and T cells (13–15). APCs, such Ni allergy, the mechanisms that regulate dermal APC responses are unknown. Therefore, using this mouse model system, we *Department of Oral Molecular Pathology, Institute of Health Biosciences, The addressed this question by determining the expression levels of University of Tokushima Graduate School, Tokushima 770-8504, Japan; and genes involved in allergy. We focused on thymic stromal lym- † Department of Oral Maxillofacial Prosthodontics, Institute of Health Biosciences, phopoietin (TSLP) and its receptor (TSLPR) because their ex- The University of Tokushima Graduate School, Tokushima 770-8504, Japan pression levels were increased significantly in the ear tissues of Received for publication January 29, 2013. Accepted for publication February 25, 2014. mice with Ni-induced allergy compared with those of other genes. Moreover, we assessed whether these findings could be used to This work was supported in part by the Funding Program for Next Generation World- Leading Researchers in Japan (LS090) and Grants-in-Aid for Scientific Research devise new therapeutic strategies or diagnostic tools to address 21792077 and 23659946 from the Ministry of Education, Culture, Sports, Science metal allergies. and Technology of Japan. The sequences presented in this article have been submitted to the Gene Expression Materials and Methods Omnibus (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE52466) under ac- cession number GSE52466. Mice Address correspondence and reprint requests to Dr. Naozumi Ishimaru, Department Female C57BL/6J mice (8-wk old) were purchased from CLEA Japan of Oral Molecular Pathology, Institute of Health Biosciences, The University of (Tokyo, Japan). Nf-kb12/2 mice were obtained from The Jackson Labo- Tokushima Graduate School, 3-18-15 Kuramotocho, Tokushima 770-8504, Japan. ratory. Mice were provided a standard laboratory diet and water. They were E-mail address: [email protected] maintained in specific pathogen–free conditions and a temperature- The online version of this article contains supplemental material. controlled environment under a 12-h light/dark cycle. The Animal Eth- Abbreviations used in this article: AD, atopic dermatitis; cLN, cervical lymph node; ics Board of the University of Tokushima approved all procedures. DC, dendritic cell; DTH, delayed-type hypersensitivity; siRNA, small interfering Induction of DTH using metals RNA; TSLP, thymic stromal lymphopoietin; TSLPR, TSLP receptor. m m This article is distributed under The American Association of Immunologists, Inc., To induce DTH with Ni, 125 l 1-mM NiCl2, with or without 125 l Reuse Terms and Conditions for Author Choice articles. IFA (ICN Biomedicals, Aurora, OH), was injected i.p. for initial immu- nization. Two weeks later, 10 ml 0.2 mM NiCl2, with or without 10 mlCFA Copyright Ó 2014 by The American Association of Immunologists, Inc. 0022-1767/14/$16.00 (ICN Biomedicals), was injected intradermally into the ear skin using a www.jimmunol.org/cgi/doi/10.4049/jimmunol.1300276 4026 A ROLE FOR TSLP IN Ni ALLERGY 26-gauge needle (TERUMO, Tokyo, Japan). DTH was determined 48 h humidified air containing 5% CO2. When the cells were 80% confluent, after the second challenge by measuring changes in ear thickness using they were stimulated with 500 or 1000 nM Ni or 1 mg/ml the TLR 4 a digital caliper. To induce DTH using other metals, 125 ml 1 mM PdCl2 or agonist LPS (Sigma Aldrich) extracted from the outer membrane of AuCl3, together with 125 ml IFA, was used for the first injection, and 10 ml Escherichia coli 0111:B4 in DMEM containing 1% FCS for 24 h. For 0.2 mM PdCl2 or AuCl3, together with 10 ml CFA, was used for the second in vitro TLR4 gene knockdown, COCA cells were transfected with TLR4 injection. or control small interfering RNAs (siRNAs; 10 nM) using Lipofectamine RNAiMAX (Life Technologies). TLR4 and negative-control siRNAs Histology (siTrio; cat no. S30C-0126) were purchased from Cosmo Bio (Tokyo, Japan). 9 Ear tissues were fixed with 10% phosphate-buffered formaldehyde (pH 7.2). The sequences of three pairs of oligonucleotides were as follows: sense, 5 - GCA UAG AGG UAG UUC CUA ATT-39; antisense, 59-UUA GGA ACU Paraffin-embedded tissue sections (4 mm) were stained with H&E. ACC UCU AUG CTT-39; sense, 59-CAC AAG AGC CGG AAG GUU ATT Flow cytometric analysis -39; antisense, 59-UAA CCU UCC GGC UCU UGU GTT-39;sense,59-GAG AAGAGGCUAAGGCCUATT-39; and antisense, 59-UAG GCC UUA Mononuclear cell preparations (1 3 106 cells)