Isolation of luteal endothelial cells and functional interactions with T lymphocytes. Issue 5 (May 2017)

• Record Type: Journal Article
• Authors: Walusimbi, S S; Wetzel, L M; Townson, D H; Pate, J L
• Is Part Of: Reproduction. Volume 153:Issue 5(2017)
• Page Start: 519
• Page End: 533
• Publication Date: 2017-05
• Subjects: Reproduction -- Periodicals; Reproduction -- Molecular aspects -- Periodicals; Reproduction -- Immunological aspects -- Periodicals; Reproduction -- Endocrine aspects -- Periodicals; Fertility -- Periodicals; Human reproduction -- Periodicals; 571.805
• DOI: 10.1530/REP-16-0578
• Languages: English
• ISSNs: 1470-1626

Abstract

Abstract : The objectives of this study were to optimize the isolation of luteal endothelial cells (LEC) and examine their functional interactions with autologous T lymphocytes. Analysis by flow cytometry showed that the purity of LEC isolated by filtration was nearly 90% as indicated by Bandeiraea simplicifolia (BS)-1 lectin binding. LEC expressed mRNA for progesterone receptor (PGR), prostaglandin receptors (PTGFR, PTGER2 and 4, and PTGIR), tumor necrosis factor receptors (TNFRSF1A&B) and interleukin (IL) 1B receptors (IL1R1&2). LEC were pretreated with either vehicle, progesterone (P4 ; 0–20 µM), prostaglandin (PG) E2 or PGF2α (0–0.2 µM), and further treated with or without TNF and IL1B (50 ng/mL each). LEC were then incubated with autologous T lymphocytes in an adhesion assay. Fewer lymphocytes adhered to LEC after exposure to high compared to low P4 concentrations (cubic response; P < 0.05). In contrast, 0.2 µM PGE2 and PGF2α each increased T lymphocyte adhesion in the absence of cytokines ( P < 0.05). LEC induced IL2 receptor alpha (CD25) expression and proliferation of T lymphocytes. In conclusion, filtration is an effective way of isolating large numbers of viable LEC. It is proposed that PGs and P4 modulate the ability of endothelial cells to bind T lymphocytes, potentially regulating extravasation, and that LEC activate T lymphocytes migrating into or resident in the CL.