Human vascular endothelial cells produce tumor necrosis factor-alpha in response to proinflammatory cytokine stimulation

Crit Care Med. 1999 Oct;27(10):2184-7. doi: 10.1097/00003246-199910000-00019.

Abstract

Objective: To determine whether human vascular endothelial cells produce tumor necrosis factor-alpha (TNF-alpha) after stimulation with proinflammatory cytokines and bacterial lipopolysaccharides (LPS).

Design: Prospective, in vitro repeated-measurements analysis of cellular responses.

Setting: Research laboratory in an academic medical center.

Subjects: Human umbilical vein endothelial cells (HUVECs).

Interventions: HUVECs were incubated with interferon-gamma (IFN-gamma), interleukin-1beta (IL-1beta), and LPS, or their different combinations for 2 to 48 hrs.

Measurements and main results: TNF-alpha was measured by time-resolved immunofluorometric assay. Unstimulated HUVECs did not produce detectable amounts of TNF-alpha, but IFN-gamma, IL-1beta, and LPS when added together induced TNF-alpha production of HUVECs in a time-dependent manner. Immunofluorescent staining confirmed that the TNF-alpha was produced by endothelial cells. IFN-gamma, IL-1beta, or LPS alone did not induce TNF-alpha production, whereas IFN-gamma and IL-1beta in combination were able to induce TNF-alpha production to some extent, and the production could be further increased with LPS. TNF-alpha messenger RNA expression was detected with reverse transcriptase-coupled polymerase chain reaction in stimulated, but not in unstimulated, HUVECs.

Conclusions: HUVECs are capable of producing TNF-alpha after proinflammatory cytokine stimulation and may therefore contribute to the increased amount of TNF-alpha found in pathologic states such as septic shock.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Count
  • Cells, Cultured
  • Culture Media
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / metabolism
  • Fluoroimmunoassay
  • Humans
  • Interferon-gamma / pharmacology
  • Interleukin-1 / pharmacology*
  • Lipopolysaccharides / pharmacology
  • Microscopy, Fluorescence
  • Prospective Studies
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Salmonella typhimurium
  • Stimulation, Chemical
  • Tumor Necrosis Factor-alpha / biosynthesis*
  • Tumor Necrosis Factor-alpha / genetics
  • Umbilical Veins / cytology

Substances

  • Culture Media
  • Interleukin-1
  • Lipopolysaccharides
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma