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Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci

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...

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Detalles Bibliográficos
Autores principales: Voelz, Kerstin, Johnston, Simon A., Rutherford, Julian C., May, Robin C.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3013146/
https://www.ncbi.nlm.nih.gov/pubmed/21209844
http://dx.doi.org/10.1371/journal.pone.0015968
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author Voelz, Kerstin
Johnston, Simon A.
Rutherford, Julian C.
May, Robin C.
author_facet Voelz, Kerstin
Johnston, Simon A.
Rutherford, Julian C.
May, Robin C.
author_sort Voelz, Kerstin
collection PubMed
description 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.
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spelling pubmed-30131462011-01-05 Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci Voelz, Kerstin Johnston, Simon A. Rutherford, Julian C. May, Robin C. PLoS One Research Article 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. Public Library of Science 2010-12-31 /pmc/articles/PMC3013146/ /pubmed/21209844 http://dx.doi.org/10.1371/journal.pone.0015968 Text en Voelz et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Voelz, Kerstin
Johnston, Simon A.
Rutherford, Julian C.
May, Robin C.
Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci
title Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci
title_full Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci
title_fullStr Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci
title_full_unstemmed Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci
title_short Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci
title_sort automated analysis of cryptococcal macrophage parasitism using gfp-tagged cryptococci
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3013146/
https://www.ncbi.nlm.nih.gov/pubmed/21209844
http://dx.doi.org/10.1371/journal.pone.0015968
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