Tumor necrosis factor-induced E-selectin expression on vascular endothelial cells.
OBJECTIVE
To determine the tumor necrosis factor (TNF) receptor type involved in induction of E-selectin expression on vascular endothelial cells.


METHODS
Prospective, in vitro repeated-measures analysis of cellular responses.


METHODS
Research laboratory in an academic medical center.


METHODS
Cultured human umbilical vein endothelial cells.


METHODS
Human umbilical vein endothelial cells were incubated with recombinant human TNF (rhTNF) to induce the expression of E-selectin on their surfaces. To block rhTNF from binding to receptors, the cells were incubated with monoclonal antibodies against TNF receptors (anti-CD120a and anti-CD120b). TNF-induced E-selectin expression of the endothelial cells, with and without blocking antibodies, was then determined using indirect immunofluorescence and flow cytometry.


RESULTS
Blocking of either CD120a or CD120b receptors individually resulted in inhibition of TNF-induced E-selectin expression on human umbilical vein endothelial cells. When both antibodies were added, the inhibition of TNF-induced E-selectin expression was synergistic. Inhibition of E-selectin expression was dependent on both TNF concentrations and antibody concentrations.


CONCLUSIONS
Both CD120a and CD120b receptors are involved in TNF-induced E-selectin expression on human umbilical vein endothelial cells. Blocking of both or one receptor type can reduce or totally inhibit expression of E-selectin on human umbilical vein endothelial cells, but the response is dependent on both TNF and antibody concentrations.
Gedeit
R G
RG
Department of Pediatrics, Medical College of Wisconsin, Children's Hospital of Wisconsin, Milwaukee 53201-1997, USA.
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002478:Cells, Cultured; D019040:E-Selectin / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D011446:Prospective Studies; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism*; D014471:Umbilical Veins
D000911:Antibodies, Monoclonal; D019040:E-Selectin; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha