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Imaging macropinosomes during Shigella infections.

Sonja Kühn1, Noelia Lopez-Montero1, Yuen-Yan Chang1

  • 1Department of Cell Biology and Infection, Institut Pasteur, Paris, France.

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Shigella bacteria hijack cellular macropinocytosis (fluid uptake) not for entry, but to create a niche. This process aids their escape into the host cell cytoplasm, crucial for infection.

Keywords:
Correlative light electron microscopyEndocytosisFluorescence imagingIntracellular pathogensMacropinocytosisShigella

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Area of Science:

  • Cell Biology
  • Microbiology
  • Pathogen-Host Interactions

Background:

  • Macropinocytosis is a cellular process for fluid uptake, utilized by various cells.
  • Pathogens frequently exploit macropinocytosis for host cell entry and infection progression.
  • Shigella, an invasive bacterial pathogen, induces macropinosome formation during epithelial cell invasion.

Purpose of the Study:

  • To investigate the role of infection-associated macropinosomes in Shigella pathogenesis.
  • To review and detail imaging techniques for analyzing macropinocytosis during pathogen entry.
  • To highlight the application of correlative microscopy for studying bacterial-induced macropinosomes.

Main Methods:

  • Utilizing genetically encoded reporters and chemical probes to track fluid-phase uptake.
  • Employing correlative light and electron microscopy (CLEM) on thin sections and large volumes.
  • Analyzing Shigella entry into epithelial cells and subsequent macropinosome dynamics.

Main Results:

  • Shigella induces macropinosomes that do not engulf bacteria but are involved in intracellular niche formation.
  • These macropinosomes facilitate vacuolar membrane rupture and bacterial release into the host cell cytosol.
  • Combined imaging approaches provide detailed characterization of Shigella-induced macropinosomes.

Conclusions:

  • Shigella manipulates host macropinocytosis for intracellular niche formation and escape, rather than direct entry.
  • Advanced imaging techniques, including CLEM, are essential for dissecting pathogen-induced macropinocytosis.
  • The findings are relevant for understanding the invasion mechanisms of other bacterial pathogens like Salmonella, Brucella, and Mycobacteria.