Rab14 regulates maturation of macrophage phagosomes containing the fungal pathogen Candida albicans and outcome of the host-pathogen interaction

Infect Immun. 2015 Apr;83(4):1523-35. doi: 10.1128/IAI.02917-14. Epub 2015 Feb 2.

Abstract

Avoidance of innate immune defense is an important mechanism contributing to the pathogenicity of microorganisms. The fungal pathogen Candida albicans undergoes morphogenetic switching from the yeast to the filamentous hyphal form following phagocytosis by macrophages, facilitating its escape from the phagosome, which can result in host cell lysis. We show that the intracellular host trafficking GTPase Rab14 plays an important role in protecting macrophages from lysis mediated by C. albicans hyphae. Live-cell imaging of macrophages expressing green fluorescent protein (GFP)-tagged Rab14 or dominant negative Rab14, or with small interfering RNA (siRNA)-mediated knockdown of Rab14, revealed the temporal dynamics of this protein and its influence on the maturation of macrophage phagosomes following the engulfment of C. albicans cells. Phagosomes containing live C. albicans cells became transiently Rab14 positive within 2 min following engulfment. The duration of Rab14 retention on phagosomes was prolonged for hyphal cargo and was directly proportional to hyphal length. Interference with endogenous Rab14 did not affect the migration of macrophages toward C. albicans cells, the rate of engulfment, the overall uptake of fungal cells, or early phagosome processing. However, Rab14 depletion delayed the acquisition of the late phagosome maturation markers LAMP1 and lysosomal cathepsin, indicating delayed formation of a fully bioactive lysosome. This was associated with a significant increase in the level of macrophage killing by C. albicans. Therefore, Rab14 activity promotes phagosome maturation during C. albicans infection but is dysregulated on the phagosome in the presence of the invasive hyphal form, which favors fungal survival and escape.

MeSH terms

  • Animals
  • Bone Marrow Cells
  • Candida albicans / immunology*
  • Candida albicans / pathogenicity
  • Cathepsins / biosynthesis
  • Cell Line
  • Green Fluorescent Proteins / genetics
  • Host-Pathogen Interactions / immunology
  • Hyphae / immunology*
  • Immune Evasion
  • Lysosomal-Associated Membrane Protein 1 / biosynthesis
  • Lysosomes / immunology
  • Lysosomes / microbiology
  • Macrophages / immunology*
  • Mice
  • Mice, Inbred C57BL
  • Phagocytosis / immunology*
  • Phagosomes / genetics
  • Phagosomes / immunology
  • Phagosomes / microbiology
  • RNA Interference
  • RNA, Small Interfering
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / immunology*
  • rab5 GTP-Binding Proteins / immunology

Substances

  • Lysosomal-Associated Membrane Protein 1
  • RNA, Small Interfering
  • Green Fluorescent Proteins
  • rab7 protein
  • Cathepsins
  • Rab14 protein, mouse
  • rab GTP-Binding Proteins
  • rab5 GTP-Binding Proteins