The current study was undertaken to investigate the role of TNF-R75 in regulation of E-selectin and ICAM-1 expression by TNF on HUVEC. To this end, we used agonistic anti-TNF-R75 antibodies, being mAb MR2-1 and polyclonal antibodies anti-TNF-R75 (pAb75). The agonistic properties of these antibodies were ascertained by the costimulatory capacity in a T-cell proliferation assay. These anti-TNF-R75 antibodies bound effectively to HUVEC, as evidenced in binding studies using 125I-TNF, but they did not induce or enhance E-selectin or ICAM-1 expression as did agonistic anti-TNF-R55 antibodies. In contrast, both MR2-1 and pAb75 inhibited specifically TNF-induced E-selectin and ICAM-1 expression, but not activation by IL-1 or LPS. These results support the hypothesis, that in cells responding to TNF via the signalling pathway of the TNF-R55, the extracellular part of TNF-R75 captures TNF and delivers it to TNF-R55, resulting in an enhanced response to TNF.