Flow cytometric evaluation of adhesion of Streptococcus pyogenes to epithelial cells

The precise roles of various surface molecules in the attachment of Streptococcus pyogenes to host epithelia are currently unclear. A flow cytometry assay that facilitates the analysis of the kinetics of S. pyogenes adhesion to epithelial cells was developed. Dose- and time-dependent adhesion isothe...

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Bibliographic Details
Published in:Journal of microbiological methods 2002-09, Vol.51 (1), p.35-42
Main Authors: Sethman, Chad R, Doyle, Ronald J, Cowan, Marjorie M
Format: Article
Language:English
Subjects:
Bacterial adhesion
Bacterial Adhesion - physiology
Bacterial diseases
Bacteriology
Biological and medical sciences
Epithelial Cells - physiology
Flow Cytometry
Fundamental and applied biological sciences. Psychology
Genetics
Human bacterial diseases
Humans
Infectious diseases
Kinetics
Kinetics of adhesion
Medical sciences
Microbiology
Mouth Mucosa - cytology
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Staphylococcal infections, streptococcal infections, pneumococcal infections
Streptococcus pyogenes
Streptococcus pyogenes - physiology
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Summary:The precise roles of various surface molecules in the attachment of Streptococcus pyogenes to host epithelia are currently unclear. A flow cytometry assay that facilitates the analysis of the kinetics of S. pyogenes adhesion to epithelial cells was developed. Dose- and time-dependent adhesion isotherms with both buccal epithelial cells (BECs) and Hep-2 cells as substrata were obtained. Although binding equilibrium is reached within 2 h on both cell types, saturation of binding sites on BECs is not achieved within a wide range of experimental conditions. This indicates a high degree of non-specific attachment to that cell type. Since no rinsing step is necessary when using flow cytometry to analyze adhesion, low-affinity associations were observable. This was confirmed by determining bacterial desorption rates early and late in the adsorption process. Binding irregularities were also easily detected since the cytometer records and displays data for up to 10,000 epithelial cells per time point. It is proposed to use this methodology to assign roles to particular surface molecules/characteristics during distinct phases of adhesion.
ISSN:0167-7012
1872-8359
DOI:10.1016/S0167-7012(02)00054-4