Stimulation of CD95-induced apoptosis in T-cells by a subtype specific neutral sphingomyelinase inhibitor

Neutral sphingomyelinase (nSMase) has been supposed to be involved in the activation of anti-apoptotic genes and, thus, could well sustain autoimmune reactions by preventing activation induced death of autoreactive T-cells. When screening cellular extracts for SMase activity in the range between pH 6.5 and 8.5 various murine tissue samples as well as cell lines of murine and human origin displayed peaks of activity, both, at pH 7.0 and 8.0. In contrast, T-cells (human T-cell lymphoma and PRA stimulated murine lymph node cells) and monocytic leukemia cells were lacking SMase activity at pH 8.0. Only one peak of activity was found at pH 7.0. Recently we described an inhibitory compound, C11AG which selectively suppresses nSMase activity. In dose-response assays using cellular extracts the pH 7.0 nSMase turned out to be almost 100-fold more sensitive to the inhibitor than the pH 8.0 nSMase. In Jurkat T-cell lymphoma cells lacking the pH 8.0 nSMase, treatment with C11AG enhanced sensitivity to apoptosis: the concentration of CD95-specific antibody anti-APO1 could be lowered by six-fold in order to induce cell death. Concomitantly the expression of the anti-apoptotic protein A1 was found to be down-regulated. In the joints of arthritic mice, apoptosis of T-cells was stimulated after application of C11AG. Accordingly, C11AG displayed curative effects on experimental arthritis: swelling and inflammation were found to be significantly alleviated. (c) 2005 Elsevier Inc. All rights reserved.