Mechanisms of IFN-γ–induced apoptosis of human skin keratinocytes in patients with atopic dermatitis

• Published in: Journal of allergy and clinical immunology Vol. 129; no. 5; pp. 1297 - 1306
• Main Authors: Rebane, Ana, Zimmermann, Maya, Aab, Alar, Baurecht, Hansjörg, Koreck, Andrea, Karelson, Maire, Abram, Kristi, Metsalu, Tauno, Pihlap, Maire, Meyer, Norbert, Fölster-Holst, Regina, Nagy, Nikoletta, Kemeny, Lajos, Kingo, Külli, Vilo, Jaak, Illig, Thomas, Akdis, Mübeccel, Franke, Andre, Novak, Natalija, Weidinger, Stephan, Akdis, Cezmi A.
• Format: Journal Article
• Language: English
• Published: New York, NY Mosby, Inc 01.05.2012; Elsevier
• Subjects: Adrenomedullin - genetics; Adrenomedullin - immunology; Adrenomedullin - metabolism; Aged; Allergic diseases; allergy; Allergy and Immunology; Annexin V; Antigens, Differentiation - genetics; Antigens, Differentiation - immunology; Antigens, Differentiation - metabolism; Apoptosis; Apoptosis - drug effects; Apoptosis - immunology; Atopic dermatitis; atopic eczema; Biological and medical sciences; Biopsy; Cells, Cultured; chemokine CCL5; Chemokine CCL5 - genetics; Chemokine CCL5 - immunology; Chemokine CCL5 - metabolism; chemokine CCL8; Chemokine CCL8 - genetics; Chemokine CCL8 - immunology; Chemokine CCL8 - metabolism; Computational Biology; Cytokine; Data processing; Dermatitis, Atopic - genetics; Dermatitis, Atopic - immunology; Dermatitis, Atopic - metabolism; Dual Specificity Phosphatase 1 - genetics; Dual Specificity Phosphatase 1 - immunology; Dual Specificity Phosphatase 1 - metabolism; Eczema; Female; fluorescent antibody technique; Fundamental and applied biological sciences. Psychology; Fundamental immunology; gamma -Interferon; Gene expression; Gene Expression Profiling; gene expression regulation; gene overexpression; genes; Genetic markers; Genetic Markers - genetics; Genome-Wide Association Study; Humans; Immunofluorescence; Immunopathology; inflammation; Inflammatory diseases; interferon-gamma; Interferon-gamma - immunology; Interferon-gamma - pharmacology; intracellular signaling peptides and proteins; Keratinocytes; Keratinocytes - drug effects; Keratinocytes - immunology; Keratinocytes - pathology; Lyn protein; Male; Medical sciences; messenger RNA; Middle Aged; Molecular modelling; mRNA expression array; NOD2 protein; Nod2 Signaling Adaptor Protein - genetics; Nod2 Signaling Adaptor Protein - immunology; Nod2 Signaling Adaptor Protein - metabolism; patients; Polymerase chain reaction; Polymorphism, Single Nucleotide; protein synthesis; reverse transcriptase polymerase chain reaction; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; Single-nucleotide polymorphism; Skin - pathology; Skin allergic diseases. Stinging insect allergies; Skin diseases; skin lesions; Up-Regulation - immunology; AD; TRAIL; TWEAK; Cytokine; qRT-PCR; allergy; FN14; atopic eczema; KORA; 7-AAD; SNP; inflammation; TNF-R; FAS; mRNA expression array; Quantitative RT-PCR; 7-Aminoactinomycin D; TNF receptor superfamily member 6; TNF-like weak inducer of apoptosis; Atopic dermatitis; TNF-related apoptosis-inducing ligand; Fibroblast growth factor–inducible 14; Cooperative Health Research in the Region of Augsburg; TNF receptor; Single nucleotide polymorphism; Human; Allergy; Immunopathology; Skin disease; Inflammation; Mechanism; Atopy; Immunology; Messenger RNA; Cell death; Eczema; Keratinocyte; Skin; Apoptosis; Gamma interferon
• ISSN: 0091-6749; 1097-6825; 1097-6825
• DOI: 10.1016/j.jaci.2012.02.020

Enhanced apoptosis of keratinocytes is the main cause of eczema and spongiosis in patients with the common inflammatory skin disease atopic dermatitis (AD). The aim of the study was to investigate molecular mechanisms of AD-related apoptosis of keratinocytes. Primary keratinocytes isolated from patients with AD and healthy donors were used to study apoptosis by using annexin V/7-aminoactinomycin D staining. Illumina mRNA Expression BeadChips, quantitative RT-PCR, and immunofluorescence were used to study gene expression. In silico analysis of candidate genes was performed on genome-wide single nucleotide polymorphism data. We demonstrate that keratinocytes of patients with AD exhibit increased IFN-γ–induced apoptosis compared with keratinocytes from healthy subjects. Further mRNA expression analyses revealed differential expression of apoptosis-related genes in AD keratinocytes and skin and the upregulation of immune system–related genes in skin biopsy specimens of chronic AD lesions. Three apoptosis-related genes (NOD2, DUSP1, and ADM) and 8 genes overexpressed in AD skin lesions (CCDC109B, CCL5, CCL8, IFI35, LYN, RAB31, IFITM1, and IFITM2) were induced by IFN-γ in primary keratinocytes. The protein expression of IFITM1, CCL5, and CCL8 was verified in AD skin. In line with the functional studies and AD-related mRNA expression changes, in silico analysis of genome-wide single nucleotide polymorphism data revealed evidence of an association between AD and genetic markers close to or within the IFITM cluster or RAB31, DUSP1, and ADM genes. Our results demonstrate increased IFN-γ responses in skin of patients with AD and suggest involvement of multiple new apoptosis- and inflammation-related factors in the development of AD.