Quorum Sensing pp 213-225 | Cite as

Assessing Pseudomonas aeruginosa Autoinducer Effects on Mammalian Epithelial Cells

  • Jake Everett
  • Rebecca Gabrilska
  • Kendra P. Rumbaugh
  • Elena Vikström
Part of the Methods in Molecular Biology book series (MIMB, volume 1673)


The human mucosal environment in the gut is rich with interactions between microbiota and mammalian epithelia. Microbes such as the Gram-negative bacterium Pseudomonas aeruginosa may use quorum sensing to communicate with other microorganisms and mammalian cells to alter gene expression. Here, we present methodologies to evaluate the effects of P. aeruginosa N-(3-oxo-dodecanoyl)-l-homoserine lactone (3O-C12-HSL) on Caco-2 cell monolayers. First, we describe a method for assessing barrier function and permeability of epithelial cells when exposed to 3O-C12-HSL by measuring transepithelial electrical resistance (TER) and paracellular flow using fluorescently labeled dextran. Secondly, we detail methods to investigate the effect of 3O-C12-HSL on protein–protein interactions of epithelial junction proteins. Lastly, we will detail imaging techniques to visualize Caco-2 barrier disruption following exposure to 3O-C12-HSL through the use of confocal laser scanning microscopy (CLSM) and a super resolution technique, stimulated emission depletion (STED) microscopy, to achieve nanoscale visualization of Caco-2 monolayers.

Key words

Host pathogen interaction Pseudomonas aeruginosa Quorum sensing Acyl homoserine lactone Transepithelial electrical resistance Paracellular permeability Cell junction IQGAP1 Imaging 


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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Jake Everett
    • 1
  • Rebecca Gabrilska
    • 1
  • Kendra P. Rumbaugh
    • 1
  • Elena Vikström
    • 2
  1. 1.Department of SurgeryTexas Tech University Health Sciences CenterLubbockUSA
  2. 2.Department of Clinical and Experimental Medicine, Faculty of Medicine and Health SciencesLinköping UniversityLinköpingSweden

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