Influence of Eap on leukocyte–endothelial-cell interactions and on the activity of NFκB. (A) Adhesion of human neutrophils to endothelial cells is shown in the absence (–) or presence of blocking mAb to CD18, ICAM-1, and CD31 as indicated (each 20 μg/mL), Eap, or protein A (each 20 μg/mL). Cell adhesion is expressed relative to control (in the absence of competitor). Data are means ± SD (n = 3) of a typical experiment; similar results were obtained in 3 separate experiments. (B) The transendothelial migration of human neutrophils in response toward 50 ng/mL MCP-1 is shown in the absence (–) or presence of mAb to CD18, mAb to ICAM-1, mAb to CD29 (each 20 μg/mL), Eap, or protein A (each 20 μg/mL). Neutrophils and inhibitors were coincubated during the whole course of the transmigration experiment (□), or neutrophils were incubated on the endothelial cells for 20 minutes in the absence of competitors in order to facilitate their initial attachment on the endothelial surface, and inhibitors were added thereafter into the wells (▪). Transmigration is presented as percent of control (in the absence of competitor). Data are means ± SD (n = 3) of a typical experiment; similar results were obtained in 3 separate experiments. (C) The DNA-binding activity of NFκB without or with TNFα or TNFα plus Eap in THP-1 cells is shown in the absence (□) or presence (▦) of ICAM-1 (10 μg/mL), as indicated. The insert demonstrates a typical EMSA for NFκB DNA-binding activity (1 indicates control; 2, TNF-α; 3, TNF-α+Eap; 4, ICAM-1; 5, ICAM-1+TNFα; and 6, ICAM-1+TNFα+Eap). (D) The expression of tissue factor without or with TNFα or TNFα plus Eap in THP-1 cells is shown in the absence (□) or presence (▦) of ICAM-1 (10 μg/mL). Densitometric data are expressed relative to control (100% control is represented in the absence of stimuli or competitors), and are means ± SD of 2 separate experiments. *P < .05.