Isolation of luteal endothelial cells and functional interactions with T lymphocytes.
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 byBandeiraea 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) E2or 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 P4concentrations (cubic response;P< 0.05). In contrast, 0.2 µM PGE2and PGF2αeach 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 P4modulate 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.
Department of Animal ScienceCenter for Reproductive Biology and Health, Pennsylvania State University, University Park, Pennsylvania, USA.
S S
SS
Walusimbi
Department of Animal ScienceCenter for Reproductive Biology and Health, Pennsylvania State University, University Park, Pennsylvania, USA.
L M
LM
Wetzel
Department of Animal and Veterinary SciencesUniversity of Vermont, Burlington, Vermont, USA.
D H
DH
Townson
Department of Animal ScienceCenter for Reproductive Biology and Health, Pennsylvania State University, University Park, Pennsylvania, USA jlp36@psu.edu.
J L
JL
Pate
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D015237:Dinoprost / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D008184:Luteal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D011374:Progesterone / Q000494:pharmacology; D011980:Receptors, Progesterone / Q000378:metabolism; D011982:Receptors, Prostaglandin / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D016207:Cytokines; D011980:Receptors, Progesterone; D011982:Receptors, Prostaglandin; D014409:Tumor Necrosis Factor-alpha; D011374:Progesterone; D015237:Dinoprost; D015232:Dinoprostone