Changes in astrocyte shape induced by sublytic concentrations of the cholesterol-dependent cytolysin pneumolysin still require pore-forming capacity

Christina Förtsch, Sabrina Hupp, Jiangtao Ma, Timothy J Mitchell, Elke Maier, Roland Benz, Asparouh I Iliev

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

Streptococcus pneumoniae is a common pathogen that causes various infections, such as sepsis and meningitis. A major pathogenic factor of S. pneumoniae is the cholesterol-dependent cytolysin, pneumolysin. It produces cell lysis at high concentrations and apoptosis at lower concentrations. We have shown that sublytic amounts of pneumolysin induce small GTPase-dependent actin cytoskeleton reorganization and microtubule stabilization in human neuroblastoma cells that are manifested by cell retraction and changes in cell shape. In this study, we utilized a live imaging approach to analyze the role of pneumolysin's pore-forming capacity in the actin-dependent cell shape changes in primary astrocytes. After the initial challenge with the wild-type toxin, a permeabilized cell population was rapidly established within 20-40 minutes. After the initial rapid permeabilization, the size of the permeabilized population remained unchanged and reached a plateau. Thus, we analyzed the non-permeabilized (non-lytic) population, which demonstrated retraction and shape changes that were inhibited by actin depolymerization. Despite the non-lytic nature of pneumolysin treatment, the toxin's lytic capacity remained critical for the initiation of cell shape changes. The non-lytic pneumolysin mutants W433F-pneumolysin and delta6-pneumolysin, which bind the cell membrane with affinities similar to that of the wild-type toxin, were not able to induce shape changes. The initiation of cell shape changes and cell retraction by the wild-type toxin were independent of calcium and sodium influx and membrane depolarization, which are known to occur following cellular challenge and suggested to result from the ion channel-like properties of the pneumolysin pores. Excluding the major pore-related phenomena as the initiation mechanism of cell shape changes, the existence of a more complex relationship between the pore-forming capacity of pneumolysin and the actin cytoskeleton reorganization is suggested.

Original languageEnglish
Pages (from-to)43-62
Number of pages20
JournalToxins
Volume3
Issue number1
DOIs
Publication statusPublished - Jan 2011

Keywords

  • Actin Cytoskeleton
  • Animals
  • Animals, Newborn
  • Astrocytes
  • Bacterial Proteins
  • Cell Membrane
  • Cell Membrane Permeability
  • Cell Shape
  • Cells, Cultured
  • Cholesterol
  • Coculture Techniques
  • Dose-Response Relationship, Drug
  • Escherichia coli
  • Image Processing, Computer-Assisted
  • Immunohistochemistry
  • Membrane Potentials
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal
  • Porosity
  • Rats
  • Rats, Sprague-Dawley
  • Spectrometry, Fluorescence
  • Streptococcus pneumoniae
  • Streptolysins
  • Virulence Factors

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