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Volume 152, Issue 9

Laboratory and clinical strains do not bind glycosphingolipids or during type IV pili-mediated initial host cell attachment Free

Abstract

The glycosphingolipids (GSLs) gangliotriaosylceramide (Gg) and gangliotetraosylceramide (Gg) have been implicated as receptors for type IV pili (T4P)-mediated epithelial cell attachment. Since T4P are divided into five groups, the authors determined whether GSLs in general, and Gg and Gg in particular, are specifically bound and required for host epithelial cell attachment of clinical and laboratory strains within these groups. An enterohaemorrhagic strain, CL56, known to bind to both Gg and Gg, provided a positive control. TLC overlay showed no binding of more than 12 strains to either Gg or Gg (or other GSLs), while CL56 Gg/Gg binding was readily detectable. GSL ELISA similarly demonstrated no significant binding to Gg or Gg, compared with CL56. Using a selective chemical inhibitor, epithelial cell GSL synthesis was abrogated, and Gg and Gg expression deleted, but attachment was not impaired. Target cell attachment was mediated by T4P, since non-piliated, but flagellated, mutants were unable to bind to the target cells. CFTR (cystic fibrosis transmembrane conductance regulator) has also been implicated as a receptor; however, in this work, overexpression of CFTR had no effect on binding. It is concluded that neither Gg nor Gg are specifically recognized by , and that endogenous GSLs do not have a role in the attachment of live intact to cultured lung epithelial cells. In contrast to whole piliated , T4P sheared from such bacteria showed significant Gg and Gg binding, which may explain the results of other studies.

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Keyword(s): CF, cystic fibrosis , CFTR, cystic fibrosis transmembrane conductance regulator , DAPI, 4′,6-diamidino-2-phenylindole dilactate , EHEC, enterohaemorrhagic E. coli , ER, endoplasmic reticulum , FBS, fetal bovine serum , Gb3, globotriaosylceramide , Gg3, gangliotriaosylceramide , Gg4, gangliotetraosylceramide , GSL, glycosphingolipid , HA, haemagglutinin , P4, 1-phenyl-2-palmitoylamino-3-pyrrolidino-1-propanol , PIBM, polyisobutylmethacrylate , RELISA, receptor ELISA , SGC, sulfagalactosylceramide , T4P, type IV pili and wt, wild-type
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2006-09-01
2024-04-19
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Laboratory and clinical Pseudomonas aeruginosa strains do not bind glycosphingolipids in vitro or during type IV pili-mediated initial host cell attachment
Microbiology 152, 2789 (2006); https://doi.org/10.1099/mic.0.28863-0
/content/journal/micro/10.1099/mic.0.28863-0
/content/journal/micro/10.1099/mic.0.28863-0
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