Skin-Infiltrating CD8+ T Cells Initiate Atopic Dermatitis Lesions Ana Hennino, Marc Vocanson, Yann Toussaint, Karen Rodet, Josette Benetière, Anne-Marie Schmitt, This information is current as Marie-Françoise Aries, Frédéric Bérard, Aurore Rozières of September 28, 2021. and Jean-François Nicolas J Immunol 2007; 178:5571-5577; ; doi: 10.4049/jimmunol.178.9.5571 http://www.jimmunol.org/content/178/9/5571 Downloaded from References This article cites 30 articles, 7 of which you can access for free at: http://www.jimmunol.org/content/178/9/5571.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 28, 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 © 2007 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Skin-Infiltrating CD8؉ T Cells Initiate Atopic Dermatitis Lesions1 Ana Hennino,*† Marc Vocanson,*† Yann Toussaint,*† Karen Rodet,*‡ Josette Benetie`re,*† Anne-Marie Schmitt,§ Marie-Franc¸oise Aries,§ Fre´de´ric Be´rard,*†‡ Aurore Rozie`res,*†‡ and Jean-Franc¸ois Nicolas2*†‡ Skin lesions in the allergic form of atopic dermatitis (AD) are induced by allergen-specific T cells that infiltrate the skin at the site of allergen exposure. Although Th2-type CD4؉ T cells appear to be crucial in AD pathophysiology, little is known about the contribution of CD8؉ T cells in the development of the allergic skin inflammation. In the present study, we have analyzed the respective role of CD8؉ and CD4؉ T cells in the development of AD skin lesions in a mouse model of allergen-induced AD. In sensitized mice, CD8؉ T cells are rapidly and transiently recruited to the allergen-exposed site and initiate the inflammatory process leading to skin infiltration with eosinophils and Th1/Th2-producing cells. CD8؉ T cell-depleted mice show no inflamma- Downloaded from tion, demonstrating that these cells are mandatory for the development of AD. In contrast, CD4؉ T cell-depleted mice develop a severe form of eczema. Furthermore, adoptive transfer of CD8؉ T cells from sensitized mice into naive recipient mice leads to skin -inflammation soon after allergen exposure. These data indicate that allergen-primed CD8؉ T cells are required for the develop ment of AD-like lesions in mice. The Journal of Immunology, 2007, 178: 5571–5577. topic diseases including atopic dermatitis (AD),3 Der p supporting the concept that immune responses in AD skin http://www.jimmunol.org/ asthma, and allergic rhinitis have a prevalence of up to are mediated by allergen-specific T cells and can be elicited by A 30% in developed countries and a high overall morbidity environmental aeroallergens (3, 5). However, the nature of T cells (1, 2). AD is a T cell-mediated chronic inflammatory skin disease responsible for the skin inflammation and the mechanisms by associated with cutaneous hyperreactivity to environmental Ags which they induce the AD skin lesions remain unknown. that are innocuous to normal nonatopic individuals. Patients with CD4ϩ T cells are considered pivotal in the development of ec- AD develop specific IgE and T cells to aeroallergens (3). In this zema and skin eosinophilic inflammation by producing a mixed respect, the house dust mites Dermatophagoides farinae (Der f) pattern of Th1 and Th2 cytokines including IL-4 and IL-13, which and Dermatophagoides pteronyssinus (Der p) are major aeroaller- are known to induce isotype switching to IgE synthesis, as well as by guest on September 28, 2021 gens in AD pathophysiology, as up to 75% of patients with AD IL-5, which plays an important role in eosinophil development and have skin prick test reactivity and/or specific IgE. More impor- survival (6, 7). Such hypothesis relies on indirect observations in tantly, epicutaneous application of house dust mite extract by human AD, mostly using peripheral blood CD4ϩ T cells and T cell atopy patch test on uninvolved skin of patients with AD elicits clones and on histological findings showing that the majority of eczema in 30–50% of these patients (3, 4). T cells isolated from skin-infiltrating T cells in active AD lesions were CD4ϩ T cells AD skin lesions and allergen patch test sites selectively respond to (8). Whereas in acute AD lesions a predominance of Th2-produc- ing cells has been observed, a shift to a Th1 cytokine pattern has been described in chronic lesions (9). Although IFN-␥ has been *Institut National de la Sante´ et de la Recherche Me´dicale, Unite´ 503, L’Institut shown to be produced in both acute and chronic skin lesions in † Fe´de´ratif de Recherche 128 BioSciences Lyon-Gerland, and Universite´Claude Ber- humans (10, 11), high numbers of CD4ϩ and CD8ϩ T cells nard Lyon 1, Lyon, France; ‡Hospices Civils de Lyon, Department of Clinical Im- munology and Allergy, Centre Hospitalier Universitaire de Lyon Sud, Pierre Be´nite, expressing IFN-␥ infiltrate acute AD lesions (12). Moreover, a France; and §Institut de Recherche Pierre Fabre, Hoˆtel Dieu St-Jacques, Toulouse, significant proportion of house dust mite-specific T cell clones France isolated from AD lesional skin were of a CD8 phenotype (13). Received for publication September 6, 2006. Accepted for publication February 6, 2007. Increased frequencies of both cutaneous lymphocyte-associated Ag (CLA)-positive CD4ϩ and CD8ϩ T cells producing type 2 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 cytokines have been detected in the blood of patients with AD with 18 U.S.C. Section 1734 solely to indicate this fact. (14). These circulating CLA-positive CD8ϩ T cells are able to 1 This work was supported by institutional grants from Institut National de la Sante´ induce IgE production by B cells and enhance eosinophils survival et de la Recherche Me´dicale, Universite´Claude Bernard Lyon 1, and Hospices Civils ´ (15). Seneviratne et al. (16) recently reported the presence of Der de Lyon and by a grant from the Centre de Recherche sur la Peau et les Epithe´lium ϩ de Reveˆtement, Pierre Fabre Laboratories (Castres, France). p-specific CD8 T cells able to produce type 1 and type 2 cyto- 2 Address correspondence and reprint requests to Dr. Jean-Franc¸ois Nicolas, Institut kines in the blood of patients with AD, but not in the blood of National de la Sante´et de la Recherche Me´dicale Unite´503, L’Institut Fe´de´ratif nonatopic individuals. More importantly these authors observed a de Recherche 128, 21 av Tony Garnier, 69375 Lyon, France. E-mail address: ϩ [email protected] or [email protected] strong correlation between the level of specific CD8 T cells and 3 the severity of the disease. Together these studies point out to a Abbreviations used in this paper: AD, atopic dermatitis; FasL, Fas ligand; CLA, ϩ cutaneous lymphocyte-associated Ag; Der f, Dermatophagoides farinae; Der p, potential role for CD8 T cells in the development of AD lesions. Dermatophagoides pteronyssinus; LN, lymph node; SFC, spot-forming cell; To examine the respective contribution of CD4ϩ and CD8ϩ T TARC, thymus- and activation-regulated chemokine. cells in AD pathophysiology, we investigated the allergic response ϩ Copyright © 2007 by The American Association of Immunologists, Inc. 0022-1767/07/$2.00 of wild-type C57BL/6 mice as compared with that of CD4 and www.jimmunol.org 5572 CD8ϩ T CELLS IN AD ϩ CD8 T cell-deficient mice, using a mouse model of allergen (Der proliferation assay. Irradiated splenocytes from naive C57BL/6 mice were f)-induced skin inflammation reproducing most of the features of used as APCs. For cytokine production (IFN-␥ and IL-4) the cells were seeded ϫ 5 ␥ human AD (17). We show that effector CD8ϩ T cells mediate at a concentration of 3 10 /well. IFN- and IL-4 production was titrated ␥ ϩ by ELISA using the R&D Systems DuoSet kit following the manufacturer inflammation and AD-like lesions. IFN- -producing CD8 T cells protocol. For ELISPOT assay (IFN-␥ and IL-5), 96-well nitrocellulose infiltrate the challenged skin a few hours after Der f exposure of plates (MAIP N4510; Millipore) were coated overnight at 4°C with anti- sensitized mice and initiate a mixed Th1/Th2-type skin inflamma- IFN-␥ Ab or anti-IL-5 Ab (R46A2; BD Pharmingen) and blocked with tion leading to the recruitment of mononuclear cells and eosino- RPMI 1640 for1hat37°C. The plates were washed five times with 0.05% PBS/Tween 20 before use. Auricular LNs of allergen- or vehicle-treated phils and to the development of epidermal changes typical of AD. mice were harvested at day 3, and cell suspensions were prepared. LN cells were dispensed in the plates (105/well) and incubated with 1 mg/ml Der f Materials and Methods proteins or Der f peptide for 36 h at 37°C, 5% CO2 in the presence of Animals irradiated splenocytes as APCs. Der f class I peptide was also used for re- stimulation at a final concentration of 2 ␮M.