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.

DESIGN

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

SETTING

Research laboratory in an academic medical center.

SUBJECTS

Cultured human umbilical vein endothelial cells.

INTERVENTIONS

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.

MEASUREMENTS AND MAIN 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.