Isolation of luteal endothelial cells and functional interactions with T lymphocytes

S. S. Walusimbi, L. M. Wetzel, D. H. Townson, J. L. Pate

Research output: Contribution to journalArticle

3 Scopus citations

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 PGF (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 PGF 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.

Original languageEnglish (US)
Pages (from-to)519-533
Number of pages15
JournalReproduction
Volume153
Issue number5
DOIs
StatePublished - Jan 1 2017

All Science Journal Classification (ASJC) codes

  • Reproductive Medicine
  • Embryology
  • Endocrinology
  • Obstetrics and Gynecology
  • Cell Biology

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