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A flow cytometry-based assay for screening FimH antagonists.

Meike Scharenberg1, Daniela Abgottspon, Evelin Cicek

  • 1Institute of Molecular Pharmacy, Pharmacenter, University of Basel, Klingelbergstrasse 50, Basel, Switzerland.

Assay and Drug Development Technologies
|June 17, 2011
PubMed
Summary

A new flow cytometry assay effectively measures how well FimH antagonists prevent uropathogenic E. coli (UPEC) from adhering to bladder cells. This method aids in developing new treatments for urinary tract infections (UTIs).

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Area of Science:

  • Microbiology
  • Urology
  • Pharmacology

Background:

  • Urinary tract infections (UTIs) are common and costly, often caused by uropathogenic Escherichia coli (UPEC).
  • UPEC adherence to the bladder epithelium, mediated by the FimH lectin on type I pili, is a critical step in UTI pathogenesis.
  • FimH antagonists, like α-d-mannopyranosides, show therapeutic potential by blocking UPEC attachment.

Purpose of the Study:

  • To develop and validate a flow cytometry-based assay for evaluating FimH antagonists.
  • To assess the inhibitory potential of various α-d-mannopyranosides against UPEC adhesion.
  • To compare the efficacy of the developed assay with existing methods.

Main Methods:

  • Utilized a human urinary bladder cell line (5637) and UPEC strain (UTI89).

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  • Employed flow cytometry to quantify UPEC adherence to uroepithelial cells.
  • Optimized and validated the assay parameters for reliable results.
  • Determined the inhibitory potency (IC50 values) of different α-d-mannopyranosides.

    • Main Results:

    • The flow cytometry assay was successfully optimized and validated for assessing FimH antagonist activity.
    • The inhibitory potencies of various α-d-mannopyranosides against UPEC adhesion were determined.
    • IC50 values obtained from the flow cytometry assay were comparable to those from other established assay formats.

    Conclusions:

    • A robust flow cytometry assay is established for evaluating FimH antagonists.
    • This assay provides a reliable method for screening potential UTI therapeutics targeting UPEC adhesion.
    • The findings support the development of FimH antagonists as a novel strategy for UTI prevention and treatment.