Measuring Antibacterial Autophagy

Methods Mol Biol. 2019:1880:679-690. doi: 10.1007/978-1-4939-8873-0_45.

Abstract

Bacteria that escape from membrane-enclosed vacuoles to the cytosol of cells are targeted by autophagy, which recognizes and captures bacteria into autophagosomes wherein their proliferation is restricted. Here we discuss two means by which antibacterial autophagy is assessed: (1) the visualization and enumeration of autophagy protein recruitment to the vicinity of cytosolic bacteria by means of immunofluorescence microscopy and (2) the measurement of autophagy-dependent restriction of bacterial proliferation by means of colony-forming unit assay.

Keywords: Colony-forming unit; Galectin; Immunofluorescence; Salmonella; Ubiquitin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagosomes / immunology*
  • Autophagosomes / microbiology
  • Autophagy / immunology*
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Colony Count, Microbial / instrumentation
  • Colony Count, Microbial / methods
  • HeLa Cells
  • Host-Pathogen Interactions / immunology*
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • RNA Interference
  • Salmonella typhimurium / genetics
  • Salmonella typhimurium / immunology
  • Salmonella typhimurium / isolation & purification
  • Transformation, Bacterial
  • Vacuoles / immunology
  • Vacuoles / microbiology

Substances

  • Luminescent Proteins
  • MAP1LC3A protein, human
  • Microtubule-Associated Proteins