Use of flow cytometry for the adhesion analysis of Streptococcus pyogenes mutant strains to epithelial cells: investigation of the possible role of surface pullulanase and cysteine protease, and the transcriptional regulator Rgg

BMC Microbiology
J HytönenJ Finne

Abstract

Flow cytometry based adherence assay is a potentially powerful but little used method in the study of bacterial binding to host structures. We have previously characterized a glycoprotein-binding activity in Streptococcus pyogenes called 'strepadhesin' binding to thyroglobulin, submaxillar mucin, fetuin and asialofetuin. We have identified surface-associated pullulanase (PulA) and cysteine protease (SpeB) as carriers of strepadhesin activity. In the present paper, we investigated the use of flow cytometry as a method to study the binding of Rgg, SpeB and PulA knock-out strains to cultured human epithelial cells. Streptococcal mutants were readily labelled with CFDA-SE and their binding to epithelial cells could be effectively studied by flow cytometry. A strain deficient in Rgg expression showed increased binding to the analyzed epithelial cell lines of various origin. Inactivation of SpeB had no effect on the adhesion, while PulA knock-out strains displayed decreased binding to the cell lines. These results suggest that the flow cytometric assay is a valuable tool in the analysis of S. pyogenes adherence to host cells. It appears to be an efficient and sensitive tool for the characterization of interactions between the bacteri...Continue Reading

References

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Citations

Dec 26, 2006·Nature Structural & Molecular Biology·Alicia Lammerts van BuerenAlisdair B Boraston
Oct 20, 2006·BMC Microbiology·Helena Tuominen-GustafssonMatti K Viljanen
Jul 15, 2015·The Journal of Infectious Diseases·Yu-Ching SuKristian Riesbeck
Oct 1, 2016·Applied Microbiology and Biotechnology·Benjamin BuysschaertNico Boon
Oct 18, 2016·Nature Microbiology·Anahita JavaheriMarkus Gerhard
Oct 12, 2018·Proceedings of the National Academy of Sciences of the United States of America·Violaine DuboisFabienne Girard-Misguich
Jun 3, 2018·The EMBO Journal·Kristof MoonensHan Remaut
Nov 15, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·A Silva-DiasC Pina-Vaz
Mar 30, 2011·Journal of Microbiological Methods·Charlotte GrootaertTom Van de Wiele

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Datasets Mentioned

BETA
GM2163

Methods Mentioned

BETA
flow cytometry
FCS
PCR

Software Mentioned

SAS
CELLQuest

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