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Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci

Lookup NU author(s): Dr Julian Rutherford

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Abstract

The human fungal pathogens Cryptococcus neoformans and C. gattii cause life-threatening infections of the central nervous system. One of the major characteristics of cryptococcal disease is the ability of the pathogen to parasitise upon phagocytic immune effector cells, a phenomenon that correlates strongly with virulence in rodent models of infection. Despite the importance of phagocyte/Cryptococcus interactions to disease progression, current methods for assaying virulence in the macrophage system are both time consuming and low throughput. Here, we introduce the first stable and fully characterised GFP-expressing derivatives of two widely used cryptococcal strains: C. neoformans serotype A type strain H99 and C. gattii serotype B type strain R265. Both strains show unaltered responses to environmental and host stress conditions and no deficiency in virulence in the macrophage model system. In addition, we report the development of a method to effectively and rapidly investigate macrophage parasitism by flow cytometry, a technique that preserves the accuracy of current approaches but offers a four-fold improvement in speed.

Publication metadata

Author(s): Voelz K, Johnston SA, Rutherford JC, May RC

Publication type: Article

Publication status: Published

Journal: PLoS ONE

Year: 2010

Volume: 5

Issue: 12

Print publication date: 31/12/2010

Date deposited: 05/05/2011

ISSN (electronic): 1932-6203

Publisher: Public Library of Science

URL: http://dx.doi.org/10.1371/journal.pone.0015968

DOI: 10.1371/journal.pone.0015968

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