Involvement of TNF-alpha receptors in the Th17 / Treg balance in pregnant women with preeclampsia

Abstract

Pregnant women with pre-eclampsia (PE) have an exacerbated immune response, with no clinical signs of infection, which can be called sterile inflammation. This activation state of innate and adaptive immunity cells results in an increase in the production of inflammatory cytokines as tumor necrosis factor alpha (TNF-alpha), and in decrease of the anti-inflammatory cytokine, interleukin-10 (IL-10). TNF-alpha can exert different functional effects on T lymphocytes through interaction with its specific receptors TNFR1 and TNFR2 present on the surface of these cells. The binding of TNF-alpha to these two receptors activates different signaling pathways. TNFR1 induces cell death by apoptosis or necrosis and is important for the development of effector T cells, while TNFR2 is predominantly expressed by CD4/Foxp3+ regulatory T cells. Co-expression of CD25 with TNFR2 identifies cells with the greatest suppressive capacity. It is already known that in PE the response of T lymphocytes is polarized to the inflammatory profile Th1 and Th17 to the detriment of the regulatory profile, and this balance between Treg and Th17 cells can be critical for the tolerance of the fetus and for the prevention of the disease. This project aims to assess the involvement of TNF-alpha receptors in the Th17/Treg balance in pregnant women with PE. Forty pregnant women will be studied, 20 with PE and 20 normotensives, paired by gestational age. The blood collected from these patients will be centrifuged, and the plasma separated and stored at -80°C for the measurement of TNF-alpha, IL-10 and the soluble forms of sTNFR1 and sTNFR2 receptors by ELISA. The determination of TNF-± receptors, TNFR1 and TNFR2 in the surface of T cell subsets, the expression of intra-cytoplasmic transcription factors RORgt (Th17) and Foxp3 (Treg) and the intracytoplasmic cytokines TNF-alpha, IL-10 and IL -17 by these cells will be evaluated by flow cytometry, using specific monoclonal antibodies marked with fluorochromes, after separation of mononuclear cells (endogenous expression). The results will be analyzed using parametric or non-parametric tests with a significance level of 5%. (AU)