DETECTION OF ADHESION OF SUPERANTIGEN-ACTIVATED T-LYMPHOCYTES TO HUMAN ENDOTHELIAL-CELLS BY ELISA

A sensitive ELISA using monoclonal antibodies (mAb) reactive with surf ace molecules specific for various leukocytes was devised to measure t he adhesion of these cells to cultured monolayers of human umbilical v ein endothelial cells. Superantigens, staphylococcal enterotoxin B or toxic shock syndrome toxin 1 were used to activate human peripheral bl ood mononuclear cells. The extent of adhesion of these cells to endoth elial 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 fix ed leukocytic cells that had adhered to endothelial cells. This method was fast and sensitive, and because the detection is by a specific ma rker on the cell of interest, it can be used in preparations of unsepa rated mixtures of cells. An increase in adhesion of superantigen-activ ated CD4(+) and CD8(+) T lymphocytes to endothelial cells may contribu te to the pathologic mechanism of superantigens.