Titan cells formation in Cryptococcus neoformans is finely tuned by environmental conditions and modulated by positive and negative genetic regulators

PLoS Pathogens
Benjamin HommelAlexandre Alanio

Abstract

The pathogenic fungus Cryptococcus neoformans exhibits morphological changes in cell size during lung infection, producing both typical size 5 to 7 μm cells and large titan cells (> 10 μm and up to 100 μm). We found and optimized in vitro conditions that produce titan cells in order to identify the ancestry of titan cells, the environmental determinants, and the key gene regulators of titan cell formation. Titan cells generated in vitro harbor the main characteristics of titan cells produced in vivo including their large cell size (>10 μm), polyploidy with a single nucleus, large vacuole, dense capsule, and thick cell wall. Here we show titan cells derived from the enlargement of progenitor cells in the population independent of yeast growth rate. Change in the incubation medium, hypoxia, nutrient starvation and low pH were the main factors that trigger titan cell formation, while quorum sensing factors like the initial inoculum concentration, pantothenic acid, and the quorum sensing peptide Qsp1p also impacted titan cell formation. Inhibition of ergosterol, protein and nucleic acid biosynthesis altered titan cell formation, as did serum, phospholipids and anti-capsular antibodies in our settings. We explored genetic factors im...Continue Reading

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Datasets Mentioned

BETA
SRR5989089
PRJNA174567

Methods Mentioned

BETA
flow cytometry
fluorescence microscopy
light microscopy
Bioscreen
FCS
with genetic modifications
PCR
electrophoresis
Illumina sequencing

Software Mentioned

ImageStreamX
RAxML
STATA®
IDEAS
ICY
ImageJ Icy
haplotypeCaller
BWA
MEM
imageJ

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