title: Methylation profile of the promoter region of IRF5 in primary Sjögren's syndrome.
abstract: The transcription factor interferon regulatory factor 5 (IRF5), in the type I interferon pathway is involved in the genetic susceptibility to various autoimmune diseases. A 5-bp insertion/deletion (CGGGG indel) polymorphism in the promoter region of IRF5 associated with primary Sjögren's syndrome (pSS) could be epigenetically deregulated in this condition. Therefore, we investigated DNA methylation patterns of the promoter region of IRF5 to determine whether its epigenetic deregulation could explain the increased expression of IRF5 mRNA in pSS patients, along with the risk of pSS induced by the genetic polymorphism. DNA extracted from total peripheral blood mononuclear cells, isolated CD4(+) T cells, B lymphocytes and monocytes from 19 pSS patients and 24 healthy controls underwent methylation analysis by pyrosequencing. Salivary gland epithelial cells (SGECs) were cultured from minor salivary glands. Regions of interest in the CGGGG repeat and ATG initiation codon region were amplified by PCR and analysed by pyrosequencing. The effect of the demethylating agent 5-AzaC on IRF5 mRNA expression in controls was quantified by RT-PCR. Among the healthy controls, the mean methylation of the nine CpG pairs of the CGGGG repeat region and the 18 CpG pairs of the ATG region was < 15% in CD4(+) T cells, B lymphocytes, monocytes and SGECs. Patients and controls did not differ in methylation profiles as regards CD4(+) T cells and B lymphocytes. IRF5 mRNA expression did not differ with or without 5-AzaC in controls. The absence of aberrant DNA methylation profiles for the putative regulatory regions of IRF5 in CD4(+) T cells, B lymphocytes, and monocytes from patients with pSS, does not support the hypothesis that epigenetic deregulation in combination with the genetic polymorphism explains the increase in IRF5 mRNA levels in pSS patients.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale INSERM U1012.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Gestermann
authors.forename: Mikael
authors.initials: M
authors.lastname: Koutero
authors.forename: Rakiba
authors.initials: R
authors.lastname: Belkhir
authors.forename: Jörg
authors.initials: J
authors.lastname: Tost
authors.forename: Xavier
authors.initials: X
authors.lastname: Mariette
authors.forename: Corinne
authors.initials: C
authors.lastname: Miceli-Richard
mesh_terms: D000328:Adult; D001374:Azacitidine / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000473:pathology; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D050835:Interferon Regulatory Factors / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012859:Sjogren's Syndrome / Q000235:genetics* / Q000276:immunology
chemical_list: C431713:IRF5 protein, human; D050835:Interferon Regulatory Factors; D012333:RNA, Messenger; D001374:Azacitidine
keywords: DNA methylation; IRF5; Sjögren's syndrome; autoimmune diseases; epigenetics