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Mechanisms of IFN-γ–induced apoptosis of human skin keratinocytes in patients with atopic dermatitis  Ana Rebane, PhD, Maya Zimmermann, PhD, Alar Aab,

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Presentation on theme: "Mechanisms of IFN-γ–induced apoptosis of human skin keratinocytes in patients with atopic dermatitis  Ana Rebane, PhD, Maya Zimmermann, PhD, Alar Aab,"— Presentation transcript:

1 Mechanisms of IFN-γ–induced apoptosis of human skin keratinocytes in patients with atopic dermatitis 
Ana Rebane, PhD, Maya Zimmermann, PhD, Alar Aab, MSc, Hansjörg Baurecht, MSc, Andrea Koreck, MD, Maire Karelson, MD, Kristi Abram, MD, Tauno Metsalu, MSc, Maire Pihlap, BSc, Norbert Meyer, MD, Regina Fölster-Holst, MD, Nikoletta Nagy, MD, PhD, Lajos Kemeny, MD, Külli Kingo, MD, PhD, Jaak Vilo, PhD, Thomas Illig, MD, Mübeccel Akdis, MD, PhD, Andre Franke, PhD, Natalija Novak, MD, Stephan Weidinger, MD, Cezmi A. Akdis, MD  Journal of Allergy and Clinical Immunology  Volume 129, Issue 5, Pages (May 2012) DOI: /j.jaci Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Increased IFN-γ–stimulated apoptosis of keratinocytes (KC) from patients with AD. A, The viability (annexin V– and 7-AAD–negative cells) of unstimulated (u.s.) or stimulated primary keratinocytes is presented as means ± SDs (n = 2). B, The viability of primary keratinocytes 4 days after stimulation (n = 5 for each group). C, One representative experiment is presented. Means ± SDs of all experiments are shown in the quadrants. *P < .05 and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Influence of IFN-γ on the expression of TRAIL, TNF-α, and their receptors in keratinocytes. A, Relative mRNA expression (mean ± SD) compared with the expression level of TNF-R2 in keratinocytes (n = 3 of each group). B, C, and E, Relative mRNA expression (mean ± SD) after stimulation with IFN-γ compared with time point 0 (=1; n = 3 from each group). D, Surface expression of the TRAIL-R2 protein (open curve) and the isotype control (solid curve). u.s., Unstimulated. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Apoptosis-related genes are differentially expressed in keratinocytes (KCs) and AD skin. A and B, The selection process (Fig 3, A) and heat map of log2 values (Fig 3, B) of differentially expressed immune system–related genes. Keratinocytes from healthy subjects (H1-H3) and patients with AD (AD1-AD3) were used. The asterisks in Fig 3, B, designate apoptosis-related genes. C, Relative mRNA expression (mean, n = 10) in lesional (L) and nonlesional (NL) AD skin is shown compared with the level seen in healthy control subjects (H, =1). *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Increased expression of IFN-γ–regulated genes in lesional biopsy specimens from patients with AD. A and B, Venn diagram of the selection process and heat maps of log2 values of selected differentially expressed genes. mRNA from lesional (L) and nonlesional (NL) skin from patients with AD and healthy control subjects (H) was analyzed. The asterisks in Fig 4, A, designate IFN-γ–regulated genes. C, Same as Fig 2, C. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 The CCL5, CCL8, and IFITM1 genes are expressed in the epidermis of skin from patients with AD. Immunofluorescence (IF), 4′,6-diamidino-2-phenylindole (DAPI), and hematoxylin (HE) staining of healthy and AD skin is shown. One representative example is shown (n = 3 for each group). Bars correspond to 10 μm (IF) and 60 μm (HE staining). The IF-stained area is designated with a square on respective HE-stained sections. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 IFN-γ response of differentially expressed genes in primary keratinocytes. A and B, Relative mRNA expression (mean ± SD, n = 6) of apoptosis-related genes (Fig 6, A) and genes differentially regulated in AD skin (Fig 6, B) in unstimulated (u.s., =1) or stimulated (24 hours) keratinocytes. C, Relative mRNA expression (mean ± SD, n = 4 for AD and healthy skin and n = 3 for psoriatic skin) after stimulation with IFN-γ compared with time point 0 (=1). *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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