title: Functionality and opposite roles of two interleukin 4 haplotypes in immune cells.
abstract: Cytokines expression can be influenced by polymorphisms in their respective coding genes. We associated the CTI/TTD haplotype (Hap-1) and TCI/CCI haplotype (Hap-2) in the IL4 gene formed by the -590, +33 and variable number of tandem repeat polymorphisms with the severity of chronic periodontitis in humans. The functionality of these IL4 haplotypes in the response of immune cells to phorbol 12-myristate 13-acetate (PMA) with Ionomycin and IL-1β (as inflammatory stimuli) was evaluated. Gene expression (quantitative real-time PCR), profile of secreted cytokines (multiplex) and phenotypic polarization of T cells (flow cytometry) were the outcomes assessed. Green fluorescent protein reporter plasmid constructs containing specific IL4 haplotype were transiently transfected into JM cells to assess the influence of the individual haplotypes on promoter activity. In response to inflammatory stimuli the immune cells from Hap-1 haplotype had increased expression of anti-inflammatory IL4; conversely, the Hap-2 haplotype showed higher levels of pro-inflammatory cytokines. The haplotype CTI proved to be the most important for the regulation of IL4 promoter, regardless of the nature of the inflammatory stimulation; whereas the polymorphism in the promoter region had the least functional effect. In conclusion, IL4 haplotypes studied are functional and trigger opposite immune responses: anti-inflammatory (Hap-1) and pro-inflammatory (Hap-2). In addition, we identified the CTI haplotype as the main responsible for the regulation of IL4 transcriptional activity.
authors.affiliation: Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP-Univ Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: G
authors.initials: G
authors.lastname: Anovazzi
authors.affiliation: Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP-Univ Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: M C
authors.initials: MC
authors.lastname: Medeiros
authors.affiliation: Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP-Univ Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: S C
authors.initials: SC
authors.lastname: Pigossi
authors.affiliation: Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP-Univ Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: L S
authors.initials: LS
authors.lastname: Finoti
authors.affiliation: Department of Biological Sciences, School of Pharmaceutical Sciences, Univ Estadual Paulista - UNESP, Araraquara, São Paulo, Brazil.
authors.forename: T M
authors.initials: TM
authors.lastname: Souza Moreira
authors.affiliation: Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: M P A
authors.initials: MP
authors.lastname: Mayer
authors.affiliation: Department of Biological Sciences, School of Pharmaceutical Sciences, Univ Estadual Paulista - UNESP, Araraquara, São Paulo, Brazil.
authors.forename: C F
authors.initials: CF
authors.lastname: Zanelli
authors.affiliation: Department of Biological Sciences, School of Pharmaceutical Sciences, Univ Estadual Paulista - UNESP, Araraquara, São Paulo, Brazil.
authors.forename: S R
authors.initials: SR
authors.lastname: Valentini
authors.affiliation: Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP-Univ Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Rossa-Junior
authors.affiliation: Department of Morphology, School of Dentistry at Araraquara, UNESP-Univ Estadual Paulista, São Paulo, Brazil.
authors.forename: R M
authors.initials: RM
authors.lastname: Scarel-Caminaga
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood*; D016022:Case-Control Studies; D055113:Chronic Periodontitis / Q000097:blood / Q000235:genetics*; D005260:Female; D005500:Follow-Up Studies; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006239:Haplotypes / Q000235:genetics*; D006801:Humans; D007249:Inflammation / Q000097:blood / Q000235:genetics*; D015847:Interleukin-4 / Q000097:blood / Q000235:genetics*; D008297:Male; D011110:Polymorphism, Genetic / Q000235:genetics*; D011379:Prognosis; D011401:Promoter Regions, Genetic / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; C508598:IL4 protein, human; D015847:Interleukin-4