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In Vitro Assay of Bacterial Adhesion onto Mammalian Epithelial Cells
05:57

In Vitro Assay of Bacterial Adhesion onto Mammalian Epithelial Cells

Published on: May 16, 2011

Galectin-3-mediated adherence of Proteus mirabilis to Madin-Darby canine kidney cells

E Altman1, B A Harrison, R K Latta

  • 1Institute for Biological Sciences, National Research Council of Canada, Ottawa, ON. Eleonora.Altman@nrc.ca

Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire
|January 22, 2002
PubMed

Insights

Proteus mirabilis uses nonagglutinating fimbriae (NAF) to adhere to kidney cells. Galectin-3 on kidney cells mediates this adherence, suggesting it

Area of Science:

  • Microbiology
  • Immunology
  • Cell Biology

Background:

  • Proteus mirabilis is a significant cause of urinary tract infections (UTIs).
  • Nonagglutinating fimbriae (NAF) of P. mirabilis mediate adherence to Madin-Darby canine kidney (MDCK) cells.
  • Understanding bacterial adherence mechanisms is crucial for combating UTIs.

Purpose of the Study:

  • To identify the specific host cell receptor involved in P. mirabilis NAF-mediated adherence to MDCK cells.
  • To elucidate the role of identified host factors in bacterial binding.

Main Methods:

  • Immunoblot overlay analysis of MDCK cell plasma membrane fractions with purified NAF.
  • Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for protein identification.
  • Immunocytochemical assays using galectin-3 specific antibodies (anti-Mac-2).

Main Results:

  • A 34-kDa protein band was identified in MDCK cell membranes that bound to NAF.
  • MALDI-TOF MS and subsequent assays identified this protein as galectin-3, located on the extracellular surface of MDCK cell membranes.
  • Antibodies against galectin-3 inhibited P. mirabilis binding to MDCK cells.

Conclusions:

  • Galectin-3 is a key mediator of P. mirabilis adherence to kidney cells in vitro.
  • The interaction involves P. mirabilis fimbriae and galectin-3, potentially with glycosylated surface receptors.
  • This finding offers insights into the pathogenesis of P. mirabilis UTIs.

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