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IL-27 Activates Th1-Mediated Responses in Imiquimod-Induced Psoriasis-Like Skin Lesions  Sayaka Shibata, Yayoi Tada, Yoshihide Asano, Koichi Yanaba, Makoto.

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Presentation on theme: "IL-27 Activates Th1-Mediated Responses in Imiquimod-Induced Psoriasis-Like Skin Lesions  Sayaka Shibata, Yayoi Tada, Yoshihide Asano, Koichi Yanaba, Makoto."— Presentation transcript:

1 IL-27 Activates Th1-Mediated Responses in Imiquimod-Induced Psoriasis-Like Skin Lesions 
Sayaka Shibata, Yayoi Tada, Yoshihide Asano, Koichi Yanaba, Makoto Sugaya, Takafumi Kadono, Naoko Kanda, Shinichi Watanabe, Shinichi Sato  Journal of Investigative Dermatology  Volume 133, Issue 2, Pages (February 2013) DOI: /jid Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 IL-27 mRNA levels and infiltration of IL-27-positive cells during the imiquimod (IMQ) application and IL-23 injection. (a) IL-27 mRNA expression levels in IMQ-induced psoriasis-like skin inflammation. Quantitative real-time PCR analysis of IL27p28 and Epstein–Barr virus–induced gene 3 (EBI3) mRNAs in the IMQ-treated skin. Total RNA was isolated from mouse back skin at day 0, 2, 4, and 6 of IMQ treatment. Data were obtained from duplicate samples from at least three mice on each day. Values are presented as means±SEs (n=6). (b) The immunohistochemical analyses for IL-27-positive cells after 6 days of IMQ treatment. The left and middle panels show IL-27 staining of intermediate-power view ( × 200) and high-power view ( × 400), respectively, in IMQ-treated skin. The right panel shows staining with isotype-matched control IgG of intermediate-power view ( × 200) in IMQ-treated skin. The arrows indicate IL-27-positive cells. The results are representative of six separate experiments. Bar=20μm. (c) Quantitative real-time PCR analysis of IL27p28 and EBI3 mRNAs in the IL-23-injected skin. Total RNA was isolated from mouse ear skin at day 0, 7, and 13 of IL-23 injection. Data were obtained from duplicate samples from at least three mice on each day. Values are presented as means±SEs (n=6). (d) The immunohistochemical analyses for IL-27-positive cells at day 13 of IL-23 injection. The left and middle panels show IL-27 staining of IL-23- and PBS-injected skin, respectively ( × 400). The right panel shows staining with isotype-matched control IgG of IL-23-injected skin ( × 200). The arrows indicate IL-27-positive cells. The results are representative of six separate experiments. Bar=20μm. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Colocalization of IL-27 with antigen-presenting cells (APCs) and dendritic cells (DCs). Imiquimod-treated skin was subjected to dual-color immunofluorescence staining for IL-27 (green, the left four panels) and major histocompatibility complex (MHC) class II, CD11c, CD3, or CD117 (red, the right four panels; × 400). The overlay images are shown in the middle four panels ( × 400). The arrows indicate the colocalization of IL-27 with MHC class II and CD11c. Bar=20μm. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 The exacerbation of imiquimod (IMQ)-induced psoriasis-like skin inflammation by IL-27 injection. (a) Phenotypical presentation (the left four panels) and histological analyses of mouse back skin (the right four panels) are shown. In each group, mice were injected with PBS or IL-27 and Vaseline or IMQ was applied. Bar=20μm. (b) Clinical scores during IMQ treatment. Erythema, scaling, and thickness of the back skin were scored on a scale from 0 to 4, and these scores were accumulated from 3 to 5 mice per group. Data are presented as means±SEs (n=5). *P<0.05 versus PBS-injected mice with IMQ application. (c) Epidermal thickness and the number of inflammatory cell infiltrates in the dermis at day 6 of IMQ treatment. Numbers of epidermal cell layers and dermal inflammatory cells were counted per high-power field from three to five mice per group. Data are presented as means±SEs (n=6). ***P<0.001 versus PBS-injected and ###P<0.001 versus IL-27-injected mice with Vaseline application, respectively. H&E, hematoxylin and eosin. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Quantitative real-time PCR analysis of control Vaseline- or imiquimod (IMQ)-treated skin with PBS or IL-27 injection. PBS- or IL-27-injected mice were treated daily with IMQ or control Vaseline on back skin, and mRNA levels were determined at day 2 by quantitative real-time PCR for T-helper 1 (Th1) cytokine/chemokines (a), Th17 cytokine/chemokines (b), and tumor necrosis factor (TNF)-α (c). Data were obtained from duplicate samples from five to eight mice in each group. Values are presented as means±SEs (n=10 to 16). *P<0.05 and **P<0.01 versus PBS-injected mice with Vaseline application, and #P<0.05 and ##P<0.01 versus IL-27-injected mice with Vaseline application, respectively. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Effects of IL-27 on the in vitro production of tumor necrosis factor (TNF)-α in peritoneal macrophages. Thioglycollate-elicited peritoneal macrophages were treated with the indicated concentrations of IL-27 alone for 24hours or were pretreated with IL-27 for 12hours, followed by treatment with lipopolysaccharide (LPS) (10ngml−1) for another 24hours. TNF-α levels in supernatants were determined using ELISA kits. Data are means±SEs (n=4) and are representative of three independent experiments. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 The improvement in imiquimod (IMQ)-induced psoriasis-like skin inflammation by IL-27 antagonism. (a) Phenotypical presentation (the left four panels) and histological analyses of mouse back skin (the right two panels) are shown. In each group, mice were injected with control IgG or anti IL-27p28 antibody (IL27p28 Ab) and IMQ was applied for 6 consecutive days. Bar=20μm. (b) Clinical scores during IMQ treatment. Erythema, scaling, and thickness of the back skin were scored on a scale from 0 to 4, and these scores were accumulated from 3 to 5 mice per group. Data are presented as means±SEs (n=5). *P<0.05 versus control IgG–injected mice with IMQ application. (c) Epidermal thickness and the number of inflammatory cell infiltrates in the dermis at day 6 of IMQ treatment. Numbers of epidermal cell layers and dermal inflammatory cells were counted per high-power field from three to five mice per group. Data are presented as means±SEs (n=6). H&E, hematoxylin and eosin. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 Effects of IL-27 antagonism on the mRNA levels of cytokine/chemokines in imiquimod (IMQ)-treated skin. Mice were intraperitoneally injected with anti-IL-27p28 antibody (IL27p28 Ab) or control IgG at day 0 and 3, and were daily treated with IMQ. mRNA levels in back skin at day 6 were determined by quantitative real-time PCR for T-helper 1 (Th1) cytokine/chemokines (a), Th17 cytokine/chemokines (b), and tumor necrosis factor (TNF)-α (c). Data were obtained from duplicate samples from five mice in each group. Values are presented as means±SEs (n=10). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

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