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*
Department of Hematology and Oncology, University of Regensburg, Regensburg, Germany;
Beatson Institute for Cancer Research, Glasgow, United Kingdom; and
Institute for Immunlogy and Cell Biology, University of Stuttgart, Stuttgart, Germany
We have previously reported that the CD14+ monocytic
subpopulation of human PBMC induces programmed cell death (apoptosis)
in cocultured endothelial cells (EC) when stimulated by bacterial
endotoxin (LPS). Apoptosis is mediated by two routes, first via
transmembrane TNF-
(mTNF) expressed on PBMC and, in addition, by
TNF-independent soluble factors that trigger apoptosis in EC.
Neutralizing anti-TNF mAb completely blocked coculture-mediated
apoptosis, despite the fact that there should have been additional
soluble cell death factors. This led to the hypothesis that a reverse
signal is transmitted from the TNF receptor on EC to monocytes (MO) via
mTNF that prevents the production of soluble apoptotic factors. Here we
have tested this hypothesis. The results support the idea of a
bidirectional cross-talk between MO and EC. Peripheral blood MO,
MO-derived macrophages (M
), or the monocytic cell line Mono Mac 6
were preincubated with human microvascular EC that constitutively
express TNF receptor type I (TNF-R1) and subsequently stimulated with
LPS. Cell-free supernatants of these preparations no longer induced EC
apoptosis. The preincubation of MO/M with TNF-reactive agents, such
as mAb and soluble receptors, also blocked the production of death
factors, providing further evidence for reverse signaling via mTNF.
Finally, we show that reverse signaling through mTNF mediated LPS
resistance in MO/M as indicated by the down-regulation of
LPS-induced soluble TNF and IL-6 as well as IL-1 and
IL-10.
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