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B Activation Induced by Bacterial Lipopolysaccharide
From the Bovine Functional Genomics Laboratory,* USDA-Agricultural Research Service, Beltsville, Maryland; and the Departments of Surgery and Medicine, University of Washington School of Medicine, Seattle, Washington
Bacterial lipopolysaccharide (LPS) via its activation of Toll-like receptor-4 contributes to much of the vascular injury/dysfunction associated with gram-negative sepsis. Inhibition of de novo gene expression has been shown to sensitize endothelial cells (EC) to LPS-induced apoptosis, the onset of which correlates with decreased expression of FLICE-like inhibitory protein (FLIP). We now have data that conclusively establish a role for FLIP in protecting EC against LPS-induced apoptosis. Overexpression of FLIP protected against LPS-induced apoptosis, whereas down-regulation of FLIP using antisense oligonucleotides sensitized EC to direct LPS killing. Interestingly, FLIP overexpression suppressed NF-
B activation induced by LPS, but not by phorbol ester, suggesting a specific role for FLIP in mediating LPS activation. Conversely, mouse embryo fibroblasts (MEF) obtained from FLIP –/– mice showed enhanced LPS-induced NF-B activation relative to those obtained from wild-type mice. Reconstitution of FLIP–/– MEF with full-length FLIP reversed the enhanced NF-B activity elicited by LPS in the FLIP –/– cells. Changes in the expression of FLIP had no demonstrable effect on other known LPS/Tlr-4-activated signaling pathways including the p38, Akt, and Jnk pathways. Together, these data support a dual role for FLIP in mediating LPS-induced apoptosis and NF-B activation.
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