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Profile of TNF-alpha receptors expressed in Th17 CD4+ lymphocytes in the peripheral blood of pregnant women with preeclampsia

Grant number: 21/12564-2
Support type:Scholarships in Brazil - Master
Effective date (Start): March 01, 2022
Effective date (End): August 31, 2023
Field of knowledge:Health Sciences - Medicine - Maternal and Child Health
Principal researcher:Maria Terezinha Serrão Peraçoli
Grantee:Patrícia Braga da Silva
Home Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Pregnant women with preeclampsia (PE) present a shift in the immune response to an inflammatory profile, resulting from the endogenous activation of innate and adaptive immunity cells. The activation of peripheral blood monocytes, with exacerbated production of inflammatory cytokines such as TNF-± and other inflammatory mediators, can cause activation of adaptive immune cells, generating inflammatory CD4+ T cells and a decrease in the production of anti-inflammatory cytokines. TNF-± can exert different functional effects on CD4+ T lymphocyte subpopulations through interaction with TNFR1 and TNFR2 receptors. The binding of TNF-± to these two receptors activates different signaling pathways. TNFR1 induces cell death by apoptosis or necrosis and is also important for the development of inflammatory effector T cells, whereas TNFR2 is preferentially expressed by regulatory T cells (Treg). It is already known that in PE, the T lymphocyte response is shifted to an inflammatory Th1 and Th17 profile in detriment to a regulatory profile, and this balance between Treg and Th17 cells may be critical for tolerance to the fetus and for disease prevention. This project aims to determine the expression of tumor necrosis factor-± receptors (TNFR1 and TNFR2) in peripheral blood CD4+ Th17 cells and its association with plasma levels of TNF-± and the soluble form of these receptors in pregnant women with PE. Forty pregnant women will be evaluated, 20 with PE and 20 normotensive pregnant women, matched for gestational age. The blood collected from these pregnant women will be centrifuged, the plasma separated and stored at -80°C for TNFR1 and TNFR2 soluble receptors, and TNF-± measurement by ELISA. The percentage of CD4+ Th17 cells will be determined by the expression of the transcription factor ROR³t, the expression of TNF-± receptors (TNFR1 and TNFR2) in this subpopulation, and the intracytoplasmic expression of IL-17A by flow cytometry, using specific monoclonal antibodies, labeled with fluorochromes soon after blood collection (endogenous expression). The results will be analyzed using parametric or non-parametric tests with a significance level of 5%. (AU)

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