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Cell Rep. 2012 Dec 27;2(6):1762-73. doi: 10.1016/j.celrep.2012.10.026. Epub 2012 Nov 29.

Tracing conidial fate and measuring host cell antifungal activity using a reporter of microbial viability in the lung.

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  • 1Vaccine and Infectious Diseases Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.

Abstract

Fluorescence can be harnessed to monitor microbial fate and to investigate functional outcomes of individual microbial cell-host cell encounters at portals of entry in native tissue environments. We illustrate this concept by introducing fluorescent Aspergillus reporter (FLARE) conidia that simultaneously report phagocytic uptake and fungal viability during cellular interactions with the murine respiratory innate immune system. Our studies using FLARE conidia reveal stepwise and cell-type-specific requirements for CARD9 and Syk, transducers of C-type lectin receptor and integrin signals, in neutrophil recruitment, conidial uptake, and conidial killing in the lung. By achieving single-event resolution in defined leukocyte populations, the FLARE method enables host cell profiling on the basis of pathogen uptake and killing and may be extended to other pathogens in diverse model host organisms to query molecular, cellular, and pharmacologic mechanisms that shape host-microbe interactions.

Copyright © 2012 The Authors. Published by Elsevier Inc. All rights reserved.

PMID:
23200858
[PubMed - indexed for MEDLINE]
PMCID:
PMC3712646
Free PMC Article
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