Biological activities and chemical composition of purified tracheal cytotoxin of Bordetella pertussis

Abstract

Specific destruction of ciliated epithelial cells lining the large airways is the primary respiratory tract cytopathology associated with human Bordetella pertussis infections. We have purified a single low-molecularweight glycopeptide, tracheal cytotoxin (TCT), that appears to cause this pathology. By using a combination of solid-phase extraction and reversed-phase high-pressure liquid chromatography, about 700 nmol of biologically active peptide can be isolated from 1 liter of B. pertussis culture supernatant (approximately 60% yield). TCT at concentrations of 1 ,uM destroyed the ciliated cell population when incubated with respiratory epithelium in vitro. This concentration of TCT is similar to the concentrations found in the culture supernatant of growing B. pertussis. Purified TCT also inhibited DNA synthesis of hamster trachea epithelial cells in a quantitative, dose-dependent fashion. Endotoxin was not detected in the purified material, and neither B. pertussis nor Escherichia coli endotoxin could duplicate the biological activities of TCT. Amino acid and amino sugar analyses of purified TCT revealed the presence of glucosamine, muramic acid, alanine, glutamic acid, and diaminopimelic acid in molar ratios of 1:1:2:1:1. This suggests that TCT, the released ciliostatic principle of B. pertussis, is a disaccharide tetrapeptide subunit of peptidoglycan.

Muramylpeptide shedding modulates cell sensing of Shigella flexneri

“…In this study we analysed the contribution of PGN sensing to the overall cellular response to Shigella infection. First, we investigated whether Shigella releases PGN fragments during PGN remodelling as originally shown for E. coli (Goodell and Schwarz, 1985) and for two human pathogens, Bordetella pertussis (Goldman et al, 1982;Cookson et al, 1989) and Neisseria gonorrhoeae (Sinha and Rosenthal, 1980). Then, we evaluated whether and how the PGN derivatives shed by Shigella could stimulate the innate immune response.…”

Peptidoglycan induces loss of a nuclear peptidoglycan recognition protein during host tissue development in a beneficial animal-bacterial symbiosis

“…Nonsymbiotic juveniles exposed to chloramphenicol were also analysed as a control for any effects of the antibiotic on EsPGRP1. For experiments involving bacterial cell envelope components, juvenile E. scolopes were exposed to 10 mM TCT and/or 10 ng ml -1 of either V. fischeri LPS or lipid A in FSIO for 24 h. Cell envelope components were purified from V. fischeri as previously described (Cookson et al, 1989;Apicella et al, 1994).…”

“…Studies of these early developmental events implicated the cell envelope constituents of V. fischeri, specifically derivatives of lipopolysaccharide (LPS) and PGN, in the triggering of host tissue morphogenesis. V. fischeri is one of the few bacterial species known to release the tetrapeptide PGN monomer, 'tracheal cytotoxin' (TCT) (Rosenthal et al, 1987;Cloud-Hansen et al, 2006), which mediates the destruction of epithelial tissues in certain pathogenic associations (Cookson et al, 1989;Cloud and Dillard, 2002). Further, as stated above, only V. fischeri can enter the crypts of E. scolopes to present these morphogens.…”

Molecular determinants of the pathogenesis of disease due to non-typable Haemophilus influenzae

“…In considering this idea, it is worth noting that Bordetella pertussis produces a virulence factor called tracheal cytotoxin that is a 921-Da muramyl-tetrapeptide derived from peptidoglycan. Goldman and colleagues have demonstrated that this molecule causes ciliostasis and extrusion of ciliated cells in hamster tracheal rings [43,44]. Neisseria gonorrhoeae produces a muramyl-tetrapeptide that is also derived from peptidoglycan and causes speci¢c damage to ciliated cells in human fallopian tube mucosa [45,46].…”