Investigation of the functional role of haplotypes in the interleukin 4 gene

• Grant number: 14/04638-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2014
• End date: March 31, 2016
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Raquel Mantuaneli Scarel Caminaga
• Grantee: Giovana Anovazzi Medeiros
• Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Periodontal Disease (PD) is multifactorial, with the influence of factors such as the presence of microorganisms periodontopathogenic, genetic susceptibility of the host, immune system reaction, smoking, stress and systemic diseases. The inflamed periodontal tissue produces various cytokines, among which interleukin 4 (IL-4). Studies by this group investigated polymorphisms in IL4 in the 250 patients with and without periodontitis, and individuals have been identified with genetic susceptibility to PD. Regarding the IL4 gene, individuals carrying the haplotype TCI/CCI formed by polymorphisms -590(T/C), +33 (T/C) and VNTR (I/D) are 5 times more susceptible to PD (ORadjusted = 5,3; 95% CI = 2,2 - 12,9), while haplotype TTD/CTI confer protection against PD ORadjusted = 0.18 (95% CI = 0,04 - 0,88). All results remained significant after adjusting for variables such as age, gender, ethnicity and smoking habit. Recently, it was investigated whether these haplotypes influence the levels of periodontal pathogens, the production of IL-4 and the response to periodontal treatment, comparing outcomes among patients carrying different haplotypes. The results suggested that individuals classified as genetically susceptible to periodontitis for carrying certain haplotypes in the IL4 gene haplotype (TCI / CCI), it was shown that individuals carrying this haplotype had higher levels of P. gingivalis, T. forsythia, T. denticola compared to patients genetically protected against DP (haplotype TTD / CTI). However, despite some significant results, it was not possible to make sure that this was due to the influence of haplotypes. Therefore, it is necessary to perform in vitro studies with more controlled conditions, to verify the actual functionality of the haplotypes in the IL4 gene. So, the objective of this study is: from cells collected of peripheral blood from patients with different haplotypes IL4 gene, investigate possible differences in innate and adaptive immune response. After conferring haplotype sequencing in gene IL4 for each patient, peripheral blood will be collected to evaluate the interaction of each polymorphism/haplotype in response to antigens by the regulation of gene expression of IL-4 (mRNA), secreted proteins (Multiplex) and intracellular protein for characterization of cell phenotypic profile. Regarding the second objective: the constructs containing the haplotype (some will pass through site-directed mutagenesis) will be transfected into NIH3T3 cells and the functionality of the different polymorphisms/haplotype in the promoter and/or intron will be assessed by reporter gene expression using fluorescence microscopy and flow cytometry. Intended to compare the response of the possible influence of different haplotypes between that obtained from cell types of patients (objective 1) the response obtained by insertion of the constructs in the lineage NIH3T3 (objective 2). (AU)

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ANOVAZZI, G.; MEDEIROS, M. C.; PIGOSSI, S. C.; FINOTI, L. S.; SOUZA MOREIRA, T. M.; MAYER, M. P. A.; ZANELLI, C. F.; VALENTINI, S. R.; ROSSA-JUNIOR, C.; SCAREL-CAMINAGA, R. M.. Functionality and opposite roles of two interleukin 4 haplotypes in immune cells. Genes and Immunity, v. 18, n. 1, p. 33-41, JAN-FEB 2017. (14/04638-2, 13/17887-8)

PIGOSSI, SUZANE C.; ANOVAZZI, GIOVANA; FINOTI, LIVIA S.; DE MEDEIROS, MARCELL C.; MAYER, MARCIA P. A.; ROSSA JUNIOR, CARLOS; SCAREL-CAMINAGA, RAQUEL M.. Functionality of the Interleukin 8 haplotypes in lymphocytes and macrophages in response to gram-negative periodontopathogens. Gene, v. 689, p. 152-160, MAR 20 2019. (14/04638-2, 13/17887-8)

PIGOSSI, SUZANE C.; ANOVAZZI, GIOVANA; FINOTI, LIVIA S.; DE MEDEIROS, MARCELL C.; DE SOUZA-MOREIRA, TATIANA MARIA; MAYER, MARCIA P. A.; ZANELLI, CLESLEI FERNANDO; VALENTINI, SANDRO ROBERTO; ROSSA JUNIOR, CARLOS; SCAREL-CAMINAGA, RAQUEL M.. The ATC/TTC haplotype in the Interleukin 8 gene in response to Gram-negative bacteria: A pilot study. ARCHIVES OF ORAL BIOLOGY, v. 107, NOV 2019. (14/04638-2, 13/17887-8)

ANOVAZZI, GIOVANA; DE MEDEIROS, MARCELL COSTA; PIGOSSI, SUZANE CRISTINA; FINOTI, LIVIA SERTORI; ALVES MAYER, MARCIA PINTO; ROSSA JUNIOR, CARLOS; SCAREL-CAMINAGA, RAQUEL MANTUANELI. Functional Haplotypes in Interleukin 4 Gene Associated with Periodontitis. PLoS One, v. 12, n. 1, JAN 23 2017. (14/04638-2, 13/17887-8)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

MEDEIROS, Giovana Anovazzi. Investigation of functionality of the haplotypes in the IL4 gene. 2016. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Faculdade de Odontologia. Araraquara Araraquara.