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Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry
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Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry

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Aspergillus fumigatus LaeA-mediated phagocytosis is associated with a decreased hydrophobin layer

Taylor R T Dagenais1, Steve S Giles, Vishukumar Aimanianda

  • 1Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, 3467 Microbial Sciences, 1550 Linden Drive, Madison, WI 53706, USA.

Infection and Immunity
|November 18, 2009
PubMed

Insights

Reduced surface rodlet protein (RodAp) on Aspergillus fumigatus laeA mutant spores increases their susceptibility to macrophage phagocytosis, a key factor in invasive aspergillosis virulence. This finding highlights RodAp

Area of Science:

  • Mycology
  • Immunology
  • Molecular Biology

Background:

  • Aspergillus fumigatus causes invasive aspergillosis, a severe disease.
  • laeA mutant spores show reduced virulence and increased phagocytosis by macrophages.
  • Key spore surface components influencing host-pathogen interactions include MAMPs, secreted metabolites, and rodlets.

Purpose of the Study:

  • To investigate the mechanisms behind the increased phagocytosis of A. fumigatus laeA mutant spores.
  • To determine the roles of cell wall MAMPs, secreted metabolites, and rodlet protein (RodAp) in macrophage-Aspergillus spore interactions.

Main Methods:

  • Flow cytometry to analyze surface-accessible MAMPs (beta-(1,3)-glucan, alpha-mannose, chitin).
  • Blocking experiments using laminarin and mannan.
  • Spent culture exchange, transwell, and coincubation assays for secreted metabolites.
  • Sonication assays, RodAp extraction/quantification, and surface layer removal to assess RodAp function.

Main Results:

  • No significant differences in surface MAMPs or carbohydrate ligands between wild-type and laeA mutant spores.
  • Secreted metabolites did not account for differential spore uptake.
  • Sonication assays and RodAp quantification indicated lower RodAp content in laeA mutant spores (60% less).
  • Removal of surface rodlets equalized phagocytosis rates between wild-type and mutant spores.

Conclusions:

  • Increased phagocytosis of laeA mutant spores is primarily due to reduced surface RodAp content, not altered MAMPs or secreted metabolites.
  • RodAp likely functions as an antiphagocytic molecule, potentially through physicochemical interactions or by hindering MAMP recognition by macrophages.

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