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 RggReportar como inadecuado




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 - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Microbiology

, 6:18

First Online: 24 February 2006Received: 28 September 2005Accepted: 24 February 2006

Abstract

BackgroundFlow 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.

ResultsStreptococcal 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.

ConclusionThese 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 bacteria and the host at the molecular level. The results also suggest a role for Rgg regulated surface molecules, like PulA, in the adhesion of S. pyogenes to host cells.

List of abbreviationsGASgroup A streptococcus

PulAstreptococcal pullulanase

SpeBstreptococcal cysteine protease

Mgamultiple gene activator of GAS

Rggregulator gene of GTF

FDSGfirst described in Streptococcus gordonii

SCCsquamous cell carcinoma

MFImean fluorescence intensity

NMFInormalized mean fluorescence intensity

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2180-6-18 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Autor: Jukka Hytönen - Sauli Haataja - Jukka Finne

Fuente: https://link.springer.com/article/10.1186/1471-2180-6-18



DESCARGAR PDF




Documentos relacionados