ST2 Regulates Allergic Airway Inflammation and T-Cell Polarization in Epicutaneously Sensitized Mice Terhi Savinko, Piia Karisola, Sari Lehtimäki, Anna-Mari.

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ST2 Regulates Allergic Airway Inflammation and T-Cell Polarization in Epicutaneously Sensitized Mice Terhi Savinko, Piia Karisola, Sari Lehtimäki, Anna-Mari Lappeteläinen, Rita Haapakoski, Henrik Wolff, Antti Lauerma, Harri Alenius Journal of Investigative Dermatology Volume 133, Issue 11, Pages 2522-2529 (November 2013) DOI: 10.1038/jid.2013.195 Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Total inflammatory cells and neutrophils are increased in the ovalbumin (OVA)/staphylococcal enterotoxin B (SEB)-sensitized skin of ST2-/- mice. Representative histological figures of (a) phosphate-buffered saline(PBS)-sensitized wild-type (WT), (b) PBS-sensitized ST2-/-, (c) OVA/SEB-sensitized WT, and (d) OVA/SEB-sensitized ST2-/- mice. Bar=200μm. (e) Total inflammatory cells, eosinophils, neutrophils, and mast cells were counted from skin samples of WT and ST2-/- mice after epicutaneous sensitization with PBS and OVA/SEB. (f) CD3+, CD4+, and CD8+ cells were counted from the epidermis and F4/80+ cells from the dermis of sensitized WT and ST2-/- mice. HPF, high-power field. *P<0.05; **P<0.01; ***P<0.001. Journal of Investigative Dermatology 2013 133, 2522-2529DOI: (10.1038/jid.2013.195) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Increased mRNA expression of IL-6, IFNγ, and IL-5 was found in the sensitized skin of ST2-/- mice as compared with wild-type (WT) mice. (a) mRNA expression of proinflammatory cytokine IL-6 and mRNA expression of Th1 cytokine IFNγ were investigated in the skin of WT and ST2-/- mice after epicutaneous sensitization with phosphate-buffered saline (PBS) and ovalbumin (OVA)/staphylococcal enterotoxin B (SEB). (b) mRNA expression of T-helper type 2 (Th2) cytokines, IL-4 and IL-5, was measured from the sensitized skin. *P<0.05; ***P<0.001. Journal of Investigative Dermatology 2013 133, 2522-2529DOI: (10.1038/jid.2013.195) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Eosinophils are downregulated and CD8+ cells are upregulated in the airways of epicutaneously sensitized and intranasally challenged ST2-/- mice. (a) Eosinophils, neutrophils, and macrophages were counted from the bronchoalveolar lavage fluid (BALF). (b) CD3+, CD4+, and CD8+ cells were stained in the lung. Lung histology was investigated from (c) phosphate-buffered saline (PBS)-sensitized wild-type (WT), (d) PBS-sensitized ST2-/-, (e) OVA/SEB-sensitized WT, and (f) OVA/SEB-sensitized ST2-/- mice. HPF, high-power field. Bar=200μm (c–f). *P<0.05; **P<0.01; ***P<0.001. Journal of Investigative Dermatology 2013 133, 2522-2529DOI: (10.1038/jid.2013.195) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 T-helper type 2 (Th2) cytokines are downregulated in the lung tissue and bronchoalveolar lavage fluid (BALF) in epicutaneously sensitized and intranasally challenged ST2-/- mice. (a) mRNA expression of tumor necrosis factor-α (TNFα), IL-13, IL-5, and IFNγ in the lung tissue of sensitized mice and controls was studied. (b) Protein concentrations of TNFα, IL-13, and IL-5 were analyzed in the BALF. (c) Foxp3 mRNA expression in the lung tissue was studied in sensitized mice. *P<0.05; **P<0.01; ***P<0.001. Journal of Investigative Dermatology 2013 133, 2522-2529DOI: (10.1038/jid.2013.195) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 IFNγ-producing CD8 + T cells are increased in the bronchoalveolar lavage fluid (BALF) of epicutaneously sensitized and intranasally challenged ST2-/- mice. (a) Ovalbumin (OVA)/staphylococcal enterotoxin B (SEB)-sensitized and OVA-challenged ST2-/- mice have increased number of CD8+ T cells that produced (b) IFNγ in the BALF compared with similarly sensitized wild-type (WT) mice. Only a low production of IFNγ was detected in CD4+ T cells. Journal of Investigative Dermatology 2013 133, 2522-2529DOI: (10.1038/jid.2013.195) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions