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Active migration into the subcellular space precedes Campylobacter jejuni invasion of epithelial cells

Authors :
Lieke B. van Alphen
Jos P. M. van Putten
Marc M. S. M. Wösten
Bas W.M. van Balkom
Nancy M. C. Bleumink-Pluym
Klazina D Rochat
Source :
Cellular Microbiology. :070725190509002
Publication Year :
2007
Publisher :
Hindawi Limited, 2007.

Abstract

The bacterial pathogen Campylobacter jejuni invades mucosal cells via largely undefined and rather inefficient (0.01-2 bacteria per cell) mechanisms. Here we report a novel, highly efficient C. jejuni infection pathway resulting in 10-15 intracellular bacteria per cell within 3 h of infection. Electron microscopy, pulse-chase infection assays and time-lapse multiphoton laser confocal microscopy demonstrated that the mechanism involved active and rapid migration of the pathogen into the subcellular space (termed 'subvasion'), followed by bacterial entry ('invasion') at the cell basis. Efficient subvasion was maximal after repeated rounds of selection for the subvasive phenotype. Targeted mutagenesis indicated that the CadF, JlpA or PEB1 adhesins were not required. Dissection of the selected and parental phenotypes by SDS-PAGE yielded comparable capsule polysaccharide and lipooligosaccharide profiles. Proteomics revealed reduced amounts of the chemotaxis protein CheW for the subvasive phenotype. Swarming assays confirmed that the selected phenotype exhibited altered migration behaviour. Introduction of a plasmid carrying chemotaxis genes into the subvasive strain yielded wild-type subvasion levels and migration behaviour. These results indicate that alterations in the bacterial migration machinery enable C. jejuni to actively penetrate the subcellular space and gain access to the cell interior with unprecedented efficiency.

Details

ISSN :
14625822 and 14625814
Database :
OpenAIRE
Journal :
Cellular Microbiology
Accession number :
edsair.doi.dedup.....4858f69cfb367b557507dea08cd1a625