Reverse signalling of membrane-integrated ligands is a common phenomenon in the tumour necrosis factor (TNF) family and contributes to the pleiotropy of this pro-inflammatory cytokine and to the plasticity of the immune system in general. Transmembrane TNF (mTNF) itself can induce resistance to bacterial endotoxin in monocytes and can stimulate the immune activity of mitogen-activated, as well as of virus-infected, T cells. The aim of the present study was to investigate the influence of reverse signalling of mTNF on the allogeneic activity of CD4+ and CD8+ T cells against human microvascular endothelial cells (HMEC), as targets of various inflammatory responses. The proliferative potential of CD4+ T cells towards HMEC was attenuated by mTNF signalling, whereas stimulation of mTNF on CD8+ T cells increased their cytotoxic potential against HMEC. These effects were specific for reverse signalling of mTNF, as a blockade of the classical TNF-TNF receptor interaction by a neutralizing TNF receptor antibody had no effect. Cytokine profiling of the effector cells revealed that the anti-endothelial CD4+ T cells were of a T helper 2 (Th2) phenotype, whereas CD8+ T cells mainly produced cytotox. T cell 1 (Tc1) cytokines. From the results obtained in this study, we conclude that reverse signalling of mTNF differentially modulates CD4+ and CD8+ T-cell activity against allogeneic endothelial cells, which should be taken into account in settings of therapeutic cytokine antagonisms.