Detection of adhesion of superantigen-activated T lymphocytes to human endothelial cells by ELISA

A sensitive ELISA using monoclonal antibodies (mAb) reactive with surface molecules specific for various leukocytes was devised to measure the adhesion of these cells to cultured monolayers of human umbilical vein endothelial cells. Superantigens, staphylococcal enterotoxin B or toxic shock syndrome toxin 1 were used to activate human peripheral blood mononuclear cells. The extent of adhesion of these cells to endothelial cells was assayed by measuring the optical density produced by a complex of peroxidase-labeled streptavidin, biotin-conjugated F(ab')(2) antimouse Ig and monoclonal antibody specific for leukocytes on fixed leukocytic cells that had adhered to endothelial cells. This method was fast and sensitive, and because the detection is by a specific marker on the cell of interest, it can be used in preparations of unseparated mixtures of cells. An increase in adhesion of superantigen-activated CD4(+) and CD8(+) T lymphocytes to endothelial cells may contribute to the pathologic mechanism of superantigens.