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.
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.
Authors: Janine L Oliver; Matthew P Alexander; Allison G Norrod; Irene M Mullins; David W Mullins Journal: Pigment Cell Melanoma Res Date: 2013-04-11 Impact factor: 4.693
Authors: Dustin R Glasner; Kalani Ratnasiri; Henry Puerta-Guardo; Diego A Espinosa; P Robert Beatty; Eva Harris Journal: PLoS Pathog Date: 2017-11-09 Impact factor: 6.823