The apoptotic response to pneumolysin is Toll-like receptor 4 dependent and protects against pneumococcal disease.

Abstract

Pneumolysin, the cholesterol-dependent cytolysin of Streptococcus pneumoniae, induces inflammatory and apoptotic events in mammalian cells. Toll-like receptor 4 (TLR4) confers resistance to pneumococcal infection via its interaction with pneumolysin, but the underlying mechanisms remain to be identified. In the present study, we found that pneumolysin-induced apoptosis is also mediated by TLR4 and confers protection against invasive disease. The interaction between TLR4 and pneumolysin is direct and specific; ligand-binding studies demonstrated that pneumolysin binds to TLR4 but not to TLR2. Involvement of TLR4 in pneumolysin-induced apoptosis was demonstrated in several complementary experiments. First, macrophages from wild-type mice were significantly more prone to pneumolysin-induced apoptosis than cells from TLR4-defective mice. In gain-of-function experiments, we found that epithelial cells expressing TLR4 and stimulated with pneumolysin were more likely to undergo apoptosis than cells expressing TLR2. A specific TLR4 antagonist, B1287, reduced pneumolysin-mediated apoptosis in wild-type cells. This apoptotic response was also partially caspase dependent as preincubation of cells with the pan-caspase inhibitor zVAD-fmk reduced pneumolysin-induced apoptosis. Finally, in a mouse model of pneumococcal infection, pneumolysin-producing pneumococci elicited significantly more upper respiratory tract cell apoptosis in wild-type mice than in TLR4-defective mice, and blocking apoptosis by administration of zVAD-fmk to wild-type mice resulted in a significant increase in mortality following nasopharyngeal pneumococcal exposure. Overall, our results strongly suggest that protection against pneumococcal disease is dependent on the TLR4-mediated enhancement of pneumolysin-induced apoptosis.

• Authors: Amit Srivastava, Philipp Henneke, Alberto Visintin, Sarah C Morse, Victoria Martin, Claire Watkins, James C Paton, Michael R Wessels, Douglas T Golenbock, Richard Malley
• Journal: Infection and immunity (Infect Immun) Vol. 73 Issue 10 Pg. 6479-87 (Oct 2005) ISSN: 0019-9567 [Print] United States
• PMID: 16177320 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Amino Acid Chloromethyl Ketones
• Bacterial Proteins
• Caspase Inhibitors
• Lipopolysaccharides
• Streptolysins
• benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
• plY protein, Streptococcus pneumoniae
• Caspases
• B 1287

Topics

• Amino Acid Chloromethyl Ketones (pharmacology)
• Animals
• Apoptosis (drug effects)
• Bacterial Proteins (metabolism, pharmacology)
• Caspase Inhibitors
• Caspases (metabolism)
• Lipopolysaccharides (pharmacology)
• Macrophages (drug effects)
• Mice
• Mice, Inbred Strains
• Nasopharynx (microbiology, pathology)
• Pneumococcal Infections (immunology, metabolism)
• Streptococcus pneumoniae (metabolism)
• Streptolysins (metabolism, pharmacology)