Macrophages are the major source of tumor necrosis factor in the porcine corpus luteum

Yulian Zhao, James A. Burbach, Katherine F. Roby, Paul F. Terranova, John D. Brannian

Research output: Contribution to journalArticlepeer-review

Abstract

This study was designed to determine the source of tumor necrosis factor (TNF) α within the porcine corpus luteum (CL). 1) Sections of frozen or paraffin-embedded CL from various stages of the estrous cycle were incubated with the following primary antibodies: anti-human recombinant TNFα, anti- porcine macrophage-specific antigen, or anti-α-actin (marker of pericyte and smooth muscle cells). Dolichos biflorus lectin-peroxidase was used as an endothelial cell label. Positive immunostaining for TNFα was apparent in porcine CL throughout the estrous cycle. TNFα immunoreactivity was primarily localized in cells along septal/vascular tracts, and exhibited spatial and temporal distribution similar to that of cells labeled with anti-macrophage antibodies. Large luteal cells exhibited weak staining for TNFα in paraffin sections, whereas microvascular endothelial cells were consistently negative in both frozen and paraffin sections. 2) Enriched subpopulations of macrophages, endothelial cells, and large and small luteal cells were isolated by density gradient and immunomagnetic bead separation techniques. TNFα secretion by each subpopulation was determined by measuring bioactive TNFα in incubation media using a specific in vitro bioassay. Macrophage subpopulations secreted up to 100-fold greater quantities of bioactive TNFα (up to 400 pg/106 cells) than did other subpopulations. In contrast, endothelial cell and small luteal cell subpopulations released very small amounts (< 8 pg/106 cells) of bioactive TNFα. Large luteal cells secreted slightly greater amounts of TNFα (10-15 pg/106 cells). Local macrophages appear to be the primary source of TNFα in the porcine CL.

Original languageEnglish (US)
Pages (from-to)1385-1391
Number of pages7
JournalBiology of reproduction
Volume59
Issue number6
DOIs
StatePublished - Dec 1998

ASJC Scopus subject areas

  • Reproductive Medicine
  • Cell Biology

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