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Official Journal of the Italian Society of Dermatology and Sexually Transmitted Diseases
Indexed/Abstracted in: EMBASE, PubMed/MEDLINE, Science Citation Index Expanded (SciSearch), Scopus
Impact Factor 1,014
Online ISSN 1827-1820
Pastore S. 1, Rogge L. 2, Mariotti F. 1, Mascia F. 1, La Scaleia R. 1, Dattilo C. 1, Sinigaglia F. 3, Girolomoni G. 4
1 Laboratory of Biochemistry Istituto Dermopatico dell'Immacolata, Roma, Italy
2 Laboratory of Immunoregulation Department of Immunology, Institut Pasteur, Paris, France
3 BioXell SpA, Milano, Italy
4 Unit of Dermatology, Department of Biomedical and Surgical Sciences, University of Verona, Verona, Italy
Aim. Keratinocytes of atopic dermatitis (AD) patients show enhanced production of cytokines and chemokines, a phenomenon that could be relevant in promoting and maintaining inflammation and hence pivotal in localizing the atopic diathesis to the skin. We performed an oligonucleotide microarrray analysis to investigate the gene expression profile in keratinocyte cultures from 6 AD patients in order to search differentially expressed genes.
Methods. Six well informed volunteer patients with moderate-to-severe chronic AD (age range 19-45 years) participated in this study. Skin involvement ranged from 20% to 60% of the body surface area. Six well informed volunteer healthy individuals (age range: 25-50 years) were used as controls.
Results. The microarrray analysis allowed to identify 201 differentially expressed transcripts, including transcriptional regulators, signal transducers, cell cycle regulators and enzymes involved in inflammation. Ten transcripts were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR), with some diverged results from gene chip analysis. In particular, metalloproteinase-2 (MMP-2) and leupaxin were not confirmed, whereas we found heterogeneous levels for c-fos, SAP1b, acidic sphingomyelinase (SMPD1), protein phosphatase 2A (PP2A) and GTP cyclohydrolase I regulatory protein among the 6 AD patients. Still, we confirmed that cathepsin O mRNA was homogeneously up-regulated in AD keratinocytes. Finally, specific transcripts of cdc2-like PISSLRE and 15-Lipooxigenase (15-LO) were undetectable as compared to controls.
Conclusion. These results indicate that the low transcript levels typical of unstimulated culture conditions may critically impair the usefulness of microarray technology in the definition of transcription profile. Furthermore, the disparate pattern of gene transcript levels in AD donors suggests that multiple and distinct molecular changes may concur to establish skin predisposition to a dysregulated response to inflammatory stimuli.