Phenotypic and functional evaluation of Regulatory T cells in gestational obesity

Abstract

During the gestational period several regulatory mechanisms are operative to maintain a homeostatic environment and to allow fetal development. In this context, regulatory T lymphocytes (Tregs) are essential for tolerance, avoiding the exacerbation of both placental and systemic inflammatory reactions. Some gestational complications, such as bacterial and viral infections, have been shown to reduce numerically and functionally the Tregs, which may lead to preterm birth and abortion. In obesity, a chronic low-grade inflammatory disease, there is a dysfunction of this cell type, leading to increased susceptibility to infections and compromised immune response. However, the role of Tregs in gestational obesity is not yet determined. This project aims to evaluate gestational obesity in mice regarding the phenotypic and functional characteristics of maternal and fetal Tregs. To this end, C57BL/6 Foxp3-GFP female mice will be induced to obesity by hypercaloric diet, monitored by weight, blood glucose and cholesterol and then mated. Tregs cells of placenta, spleen and adipose tissue of pregnant obese mice and spleen Tregs of their neonates will be evaluated. Treg cell subtypes will be evaluated for transcription factors (Foxp3, T-bet, GATA3, RORg-t) by flow cytometry and serum adipokine, leptin and cytokines IL-10 and TGF-b levels. The expression profile of Foxp3, IL-10, TGF-b, TNF and IL-1b will be evaluated by real time PCR in the placental tissue. The functional capacity of Tregs will be evaluated by cell suppression assay. The development of this project may help to understand the regulatory mechanisms in the maternal-fetal interface mediated by Tregs cells in metabolic complications such as gestational obesity and its impact on neonates.