Effect of tumor necrosis factor (TNF) on activation and expression of receptors TNFR1 and TNFR2 in lymphocytes subsets Th17 and Treg from healthy women

• Grant number: 24/05453-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2024
• End date: November 30, 2024
• Field of knowledge: Biological Sciences - Immunology - Applied Immunology
• Principal Investigator: Maria Terezinha Serrão Peraçoli
• Grantee: Giovana Checco
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Introduction: Pre-eclampsia (PE) is a pregnancy-specific syndrome characterized by high plasma levels of pro-inflammatory cytokines, such as tumor necrosis factor (TNF), which play an important role in the pathophysiology of the disease. The binding of TNF to the TNFR1 and TNFR2 receptors on the cell surface activates different signaling pathways. TNFR1 induces cell death by apoptosis or necrosis, and is also important for the development of effector T cells, while TNFR2 is predominantly expressed by CD4+/FoxP3+ regulatory T cells (Treg), and the co-expression of CD25 with TNFR2 identifies cells with the greatest suppressor capacity. Although the imbalance of the Treg/Th17 relationship has already been well described in PE, the understanding of TNF role in modulating the TNFR1 and TNFR2 receptors in T cell subsets will bring new knowledge to the understanding of the systemic inflammatory response as well as in the imbalance of T cell subsets in this pathology. Objective: This study aims to evaluate the role of soluble recombinant TNF on the activation of CD4+ T cell subsets (Th17 and Treg) from healthy non-pregnant women by inducing TNF receptors in the membrane of these cells. Subjects and Methods: Peripheral blood from 16 healthy, non-pregnant women will be obtained and mononuclear cells (PBMCs) will be cultured in vitro in the presence or absence of TNF for 4h and 18h. The cell membrane expression of TNFR1 and TNFR2 will be determined by flow cytometry. The concentration of soluble TNF receptors (sTNFR1 and sTNFR2) will be performed in the culture supernatant of these cells by enzyme immunoassay (ELISA). The results will be analyzed using parametric or non-parametric tests with a significance level of 5%.