Pasteurella multocida enters polarized epithelial cells by interacting with host F-actin

We investigated the interaction of an avian strain of Pasteurella multocida with the cytoskeleton of MDCK cells, which formed a polarized epithelium when grown on type I collagen coated filters. Bacteria were incubated with MDCK cells for 30 min, 2, 4 and 6 hours and their location and association w...

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Bibliographic Details
Published in:Veterinary microbiology 1997-03, Vol.54 (3), p.343-355
Main Authors: Rabier, Mireille J., Tyler, Nancy K., Walker, Naomi J., Hansen, Lori M., Hirsh, Dwight C., Tablin, Fern
Format: Article
Language:English
Subjects:
Actin
Actin Cytoskeleton - microbiology
Actin Cytoskeleton - ultrastructure
Actins - physiology
Animals
Antibodies, Bacterial
Bacteriology
Biological and medical sciences
Cell Line
Confocal microscopy
Cytochalasin D
Cytoskeleton
Dogs
Epithelium - microbiology
Epithelium - physiology
Fundamental and applied biological sciences. Psychology
Kidney
Microbiology
Microscopy, Confocal
Microscopy, Electron
Microvilli - microbiology
Microvilli - ultrastructure
Pasteurella multocida
Pasteurella multocida - physiology
Pasteurella multocida - ultrastructure
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
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Summary:We investigated the interaction of an avian strain of Pasteurella multocida with the cytoskeleton of MDCK cells, which formed a polarized epithelium when grown on type I collagen coated filters. Bacteria were incubated with MDCK cells for 30 min, 2, 4 and 6 hours and their location and association with the cell cytoskeleton determined by double-label immunofluorescence confocal microscopy. Cells were stained with a polyclonal antiserum to the outer-membrane proteins of P. multocida and with rhodamine phalloidin which specifically binds filamentous (F) actin. Confocal microscopy revealed that bacteria entered the cells by 30 min, and that by 6 hours there was a marked alteration in the actin cytoskeleton in which long filaments were reorganized to discrete foci of short actin filaments, within which were one or more bacteria. Electron microscopy demonstrated that by 2 hours, each bacterium was associated with many short 5–6 nm filaments. Treatment of MDCK cells with cytochalasin D for either 30 minutes or 24 hours prior to infection disrupted the actin cytoskeleton and inhibited entry of P. multocida.
ISSN:0378-1135
1873-2542
DOI:10.1016/S0378-1135(96)01255-2