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Gene Expression Profiling of Lichen Planus Reflects CXCL9+-Mediated Inflammation and Distinguishes this Disease from Atopic Dermatitis and Psoriasis 

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Presentation on theme: "Gene Expression Profiling of Lichen Planus Reflects CXCL9+-Mediated Inflammation and Distinguishes this Disease from Atopic Dermatitis and Psoriasis "— Presentation transcript:

1 Gene Expression Profiling of Lichen Planus Reflects CXCL9+-Mediated Inflammation and Distinguishes this Disease from Atopic Dermatitis and Psoriasis  Joerg Wenzel, Bettina Peters, Sabine Zahn, Michael Birth, Kay Hofmann, Daniel Küsters, Stefan Tomiuk, Jens M. Baron, Hans F. Merk, Cornelia Mauch, Thomas Krieg, Thomas Bieber, Thomas Tüting, Andreas Bosio  Journal of Investigative Dermatology  Volume 128, Issue 1, Pages (January 2008) DOI: /sj.jid Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Comparative results of SAGE analyses of whole skin samples in different inflammatory skin disorders. SAGE analyses were performed with pooled mRNA, which was extracted from skin samples of 20 patients with lichen planus (LP), atopic dermatitis (AD), psoriasis (Pso), and healthy controls (HC), respectively. In each pool, approximately 12,000–13,000 tags were identified. (a) VENN diagram of genes, which were significantly differentially expressed (at P<0.01) in the investigated skin disorders, when compared with HC. Subsequently, these tags were classified following the gene ontology clusters. (b) Overview about gene clusters which were specifically found in LP, compared with the expression in AD and Pso. Journal of Investigative Dermatology  , 67-78DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 PIQOR microarray assays reveal type I IFN signature and strong expression of CXCL9 in LP. cDNA microarray-based assays (PIQOR), including 1,542 skin-associated genes, were performed to confirm the SAGE results on an individual basis. (a) The results of unclassified clustering which clearly distinguished LP and HC from AD and Pso. (b) Examples of IFN-associated genes (chemokine ligand CXCL9, type I IFN-inducible Mx-protein, IFN regulating factor 1, and HLA molecules DPB1 and C), which were predominantly upregulated in LP (given is the relative expression when compared with a common reference which represents a pool of 160 samples derived from skin biopsies of patients with different skin disorders). Journal of Investigative Dermatology  , 67-78DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 ISH and IHC confirm significant CXCL9 expression in LP skin lesions. ISH and IHC for CXCL9 were established to confirm lesional CXCL9 expression in LP on the mRNA and the protein level. (a) Typical findings of CXCL9 ISH in LP compared with sense control (Bar=0.1mm), (b) an overview of overall investigated samples (n=5). (c and d) These results were confirmed by IHC, typical findings of CXCL9 expression in LP, AD, Pso, and HC concludes data of all investigated biopsies (n≥5, respectively). Statistical data is given in mean±SEM (*=P<0.05, **=P<0.01, Mann–Whitney U-test. Bar=0.1mm). Journal of Investigative Dermatology  , 67-78DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Type I IFNs (IFN-α/IFN-β) are largely produced in LP lesions. Lesional type I IFN signaling was investigated by ISH for IFN-α1 and IFN-β. (a) Representative findings for these ISHs in comparison with sense control (Bar=0.1mm). (b) Results of all these experiments are concluded (n=5). (c and d) The M × A protein, which is an established and highly specific marker for type I IFN production, was stained by IHC. Typical findings in the investigated skin disorders give an overview about all investigated samples (n≥5, respectively). Statistical data is given in mean±SEM (*P<0.05, **P<0.01, Mann–Whitney U-test). Journal of Investigative Dermatology  , 67-78DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Characterization of the inflammatory infiltrate by IHC reveals CD8-dominated T-cellular inflammation in LP, including numerous CXCR3+ cytotoxic cells. IHC using monoclonal antibodies specific for CD3, CD4, CD8, CD20, CD68, CXCR3, granzyme B, and Tia1 was performed to characterize the lesional infiltrate in the investigated inflammatory skin disorders. (a) Typical findings of CD8 and CXCR3 expression (Bar=0.1mm) and (b) concludes this data (n≥10, respectively). Given is the mean number of cells per high-power field (Original magnification × 200)±SEM. (c) Sequential double staining was performed to confirm coexpression of CXCR3 (brown) and granzyme B (red) in infiltrating cells. Journal of Investigative Dermatology  , 67-78DOI: ( /sj.jid ) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

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