Escherichia coli K1 invasion of human brain microvascular endothelial cells

Methods Enzymol. 2012:506:93-113. doi: 10.1016/B978-0-12-391856-7.00030-5.

Abstract

The pathogenic Escherichia coli strain E. coli K1 is a primary causative agent of neonatal meningitis. Understanding how these bacteria cross the blood-brain barrier is vital to develop therapeutics. Here, we describe the use of live-cell imaging techniques to study E. coli K1 interactions with cellular markers following infection of human brain microvascular endothelial cells, a model system of the blood-brain barrier. We also discuss optimization of endothelial cell transfection conditions using nonviral transfection technique, bacterial labeling techniques, and in vitro assays to screen for fluorescent bacteria that retain their ability to invade host cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood-Brain Barrier / microbiology
  • Brain / blood supply
  • Brain / microbiology*
  • Cell Survival
  • Endothelium, Vascular / microbiology*
  • Escherichia coli / genetics
  • Escherichia coli / pathogenicity
  • Escherichia coli / physiology*
  • Escherichia coli Infections / microbiology*
  • Fluorescent Dyes / analysis
  • Gene Expression
  • Host-Parasite Interactions*
  • Humans
  • Luminescent Proteins / analysis
  • Luminescent Proteins / genetics
  • Microscopy, Fluorescence / methods*
  • Transfection / methods

Substances

  • Fluorescent Dyes
  • Luminescent Proteins