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Background  Probiotics appear to be beneficial in inflammatory bowel disease, but their mechanism of action is incompletely understood. We investigated whether probiotic-derived sphingomyelinase mediates this beneficial effect.    Methodology/Principal Findings  Neutral sphingomyelinase (NSMase) activity was measured in sonicates of the probiotic  L. brevis (LB)  and  S. thermophilus (ST)  and the non-probiotic  E. coli (EC)  and  E. faecalis (EF) . Lamina propria mononuclear cells (LPMC) were obtained from patients with Crohn's disease (CD) and Ulcerative Colitis (UC), and peripheral blood mononuclear cells (PBMC) from healthy volunteers, analysing LPMC and PBMC apoptosis susceptibility, reactive oxygen species (ROS) generation and JNK activation. In some experiments, sonicates were preincubated with GSH or GW4869, a specific NSMase inhibitor. NSMase activity of  LB  and  ST  was 10-fold that of  EC  and  EF  sonicates.  LB  and  ST  sonicates induced significantly more apoptosis of CD and UC than control LPMC, whereas  EC  and  EF  sonicates failed to induce apoptosis. Pre-stimulation with anti-CD3/CD28 induced a significant and time-dependent increase in  LB -induced apoptosis of LPMC and PBMC. Exposure to  LB  sonicates resulted in JNK activation and ROS production by LPMC. NSMase activity of  LB  sonicates was completely abrogated by GW4869, causing a dose-dependent reduction of  LB -induced apoptosis.  LB  and  ST  selectively induced immune cell apoptosis, an effect dependent on the degree of cell activation and mediated by bacterial NSMase.    Conclusions  These results suggest that induction of immune cell apoptosis is a mechanism of action of some probiotics, and that NSMase-mediated ceramide generation contributes to the therapeutic effects of probiotics.

Probiotic Sonicates Selectively Induce Mucosal Immune Cells Apoptosis through Ceramide Generation via Neutral Sphingomyelinase