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Circadian Gene Clock Regulates Psoriasis-Like Skin Inflammation in Mice  Noriko Ando, Yuki Nakamura, Rui Aoki, Kayoko Ishimaru, Hideoki Ogawa, Ko Okumura,

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Presentation on theme: "Circadian Gene Clock Regulates Psoriasis-Like Skin Inflammation in Mice  Noriko Ando, Yuki Nakamura, Rui Aoki, Kayoko Ishimaru, Hideoki Ogawa, Ko Okumura,"— Presentation transcript:

1 Circadian Gene Clock Regulates Psoriasis-Like Skin Inflammation in Mice 
Noriko Ando, Yuki Nakamura, Rui Aoki, Kayoko Ishimaru, Hideoki Ogawa, Ko Okumura, Shigenobu Shibata, Shinji Shimada, Atsuhito Nakao  Journal of Investigative Dermatology  Volume 135, Issue 12, Pages (December 2015) DOI: /jid Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Clock mutation results in reduced imiquimod (IMQ)-induced skin inflammation. (a) Psoriasis Area Severity Index (PASI) score in the dorsal skin (n=10–12 per group). (b) Ear thickness (n=10–12 per group). (c) Hematoxylin and eosin (HE) staining of the dorsal skin (n=5 per group). Bar=100μm. (d) Frequency of γ/δ+ T cells (CD3+γ/δ+ cells, IL-22+γ/δ+ cells, CD3+ IL-22+ cells) in the cervical lymph nodes evaluated by FACS analysis at Zeitgeber time 2 (ZT2) (n=7 per group). (e) IL-22, IL-17A, S100A8, Reg3γ, IL-23 p19, and keratin 16 mRNA expression at ZT2 in the inflamed skin evaluated by quantitative reverse transcriptase in real time (qPCR) (n=3–5 per group). Amplification was normalized to GAPDH. (f) Frequency of I-A/I-E+IL-23+ cells in the cervical lymph nodes evaluated by FACS analysis at ZT2 (n=3–4 per group). The values represent means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Clock mutation blunts γ/δ+ T cell responses to IL-23. (a) IL-23-induced IL-22 and IL-17A production in splenic γ/δ+ T cells isolated from wild-type or ClockΔ19/Δ19 mice detected by ELISA (n=3 per group). (b and c) Wild-type or ClockΔ19/Δ19 mice underwent intradermal injection of phosphate-buffered saline (PBS) or IL-23 in the ears on days 1, 3, and 5 at Zeitgeber time 2 (ZT2). (b) Ear thickness (n=4–5 per group) or (c) hematoxylin and eosin (HE) staining of the ear and its quantitative analysis (epidermal thickness) (n=4–5 per group) on day 6 are shown. Bar=100μm. The values represent means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Clock regulates IL-23 receptor (IL-23R) expression in γ/δ T cells. (a) IL-23 receptor (IL-23R) mRNA expression in splenic γ/δ+ T cells isolated from wild-type mice or ClockΔ19/Δ19 mice (n=3 per group). (b) Frequency of splenic γ/δ+IL-23R+ cells isolated from wild-type mice or ClockΔ19/Δ19 mice (n=4 per group). (c) Detection of CLOCK binding to the promoter region of IL-23R in splenic γ/δ+ T cells isolated from wild-type mice evaluated by chromatin immunoprecipitation (ChIP) assay. The values represent means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 IL-23 receptor (IL-23R) expression shows circadian rhythms in the mouse skin. (a) Whole-skin samples were obtained from wild-type mice at the indicated time points, mRNA was extracted, and quantitative reverse transcriptase in real time (qPCR) analysis was performed for IL-23R, Period2 (Per2), Clock, and Bmal1(Arntl) mRNA. The values represent the means±SD (n=4–5 per group). *P<0.05. (b–d) Wild-type mice were treated with imiquimod (IMQ) for 5days (days 1–5) either at Zeitgeber time 2 (ZT2) or at ZT14. (b) Ear thickness (n=5 per group), (c) epidermal thickness (n=5 per group), or (d) frequency of CD3+γ/δ+ cells in the cervical lymph nodes evaluated by FACS analysis (n=5 per group) on day 6 is shown. The values represent the means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Period2 (Per2) mutation enhances imiquimod (IMQ)-induced skin inflammation. (a) Psoriasis Area Severity Index (PASI) score in the dorsal skin (n=3–4 per group). (b) Ear thickness (n=3–4 per group). (c) Frequency of γ/δ T cells (CD3+γ/δ+, IL-22+γ/δ+s, and CD3+IL-22+ cells) in the cervical lymph nodes evaluated by FACS analysis on day 6 (n=3 per group). (d) IL-23 receptor (IL-23R) mRNA expression evaluated by quantitative reverse transcriptase in real time (qPCR) in splenic γ/δ+ T cells isolated from nontreated wild-type mice or mPer2m/m mice (n=3 per group). (e) Frequency in splenic γ/δ+IL-23R+ cells isolated from nontreated wild-type or mPer2m/m mice evaluated by FACS analysis at Zeitgeber time 2 (ZT2) (n=3 per group). (fand g) Wild-type mice or mPer2m/m mice underwent intradermal injection of phosphate-buffered saline (PBS) or IL-23 in the ears on days 1, 3, 5, 7, and 9 at ZT2. (f) Ear thickness (n=3 per group) or (g) hematoxyin and eosin (HE) staining of the ear and its quantitative analysis (epidermal thickness) (n=3 per group) on day 10 are shown. Bar=100μm. The values represent the means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

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