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High glucose and function of dendritic cells: effects on proliferation, survival, activation and phagocytosis

Grant number: 15/07932-1
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2015
Effective date (End): May 31, 2016
Field of knowledge:Health Sciences - Dentistry - Periodontology
Principal researcher:Carlos Rossa Junior
Grantee:Debora Emy Miyazaki Lopes
Home Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Dendritic cells are the prototypical antigen-presenting cells and, as such, represent an essential link between innate and adaptive immune response. Thus, dendritic cells play a fundamental role in infectious conditions, such as periodontal diseases. Moreover, these cells also have a prominent role in allergic, autoimmune and metabolic conditions associated with chronic inflammation, including asthma, psoriasis, chronic obstructive pulmonary diseases and diabetes. Diabetes is associated with major social and economic costs, as a condition with increasing prevalence and incidence, which is associated with high mortality and morbidity due to the complications involving vascular, nervous and inflammatory changes. The association between poorly-controlled diabetes and increased extent and severity of destructive periodontal disease is widely documented and acknowledged; however the biological mechanisms involved are not well understood. Considering: 1) the de-regulation of immune response associated with diabetes; 2) the fundamental role of dendritic cells in the immune response; 3) the association between diabetes and destructive periodontal diseases and 4) the fundamental role of the immune response in destructive periodontal diseases we propose the following hypothesis for this project: high glucose concentration, the hallmark feature of diabetes, affects the biology of dendritic cells. To test this hypothesis, we propose in vitro experiments using a murine cell line of dendritic cells, which we will expose to high glucose concentration for 3 and 5 days and subsequently assess the following outcomes of interest: cell proliferation (by direct cell counting on an automated cell counter), cell survival (by Annexin V/ nuclear membrane permeability flow cytometry-based assay), dendritic cell activation/maturation (by assessing expression of specific markers using the flow cytometer after activation with bacterial LPS) and phagocytosis (using fluorescent-labeled bacterial particles and detection using flow cytometer or fluorescent microscopy).

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