Hyphal Growth of Phagocytosed Fusarium oxysporum Causes Cell Lysis and Death of Murine Macrophages

Schäfer, Katja; Bain, Judith M.; Pietro, Antonio Di; Gow, Neil A. R.; Erwig, Lars P.

doi:10.1371/journal.pone.0101999

Cited by 10 publications

(9 citation statements)

References 37 publications

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“…Aborted mitosis of the host cell after the uptake of C. albicans has been reported previously (Lewis et al, 2012b ; Schafer et al, 2014 ). After the engulfment of fungal cells, the phagocytic cell starts to divide but instead of separating, the two attached daughter cells fuse back together.…”

Section: Resultssupporting

confidence: 58%

Kinetic studies of Candida parapsilosis phagocytosis by macrophages and detection of intracellular survival mechanisms

Tóth

Tóth²,

Papp

et al. 2014

Front. Microbiol.

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Even though the number of Candida infections due to non-albicans species like C. parapsilosis has been increasing, little is known about their pathomechanisms. Certain aspects of C. parapsilosis and host interactions have already been investigated; however we lack information about the innate cellular responses toward this species. The aim of our project was to dissect and compare the phagocytosis of C. parapsilosis to C. albicans and to another Candida species C. glabrata by murine and human macrophages by live cell video microscopy. We broke down the phagocytic process into three stages: macrophage migration, engulfment of fungal cells and host cell killing after the uptake. Our results showed increased macrophage migration toward C. parapsilosis and we observed differences during the engulfment processes when comparing the three species. The engulfment time of C. parapsilosis was comparable to that of C. albicans regardless of the pseudohypha length and spatial orientation relative to phagocytes, while the rate of host cell killing and the overall uptake regarding C. parapsilosis showed similarities mainly with C. glabrata. Furthermore, we observed difference between human and murine phagocytes in the uptake of C. parapsilosis. UV-treatment of fungal cells had varied effects on phagocytosis dependent upon which Candida strain was used. Besides statistical analysis, live cell imaging videos showed that this species similarly to the other two also has the ability to survive in host cells via the following mechanisms: yeast replication, and pseudohypha growth inside of phagocytes, exocytosis of fungal cells and also abortion of host cell mitosis following the uptake. According to our knowledge this is the first study that provides a thorough examination of C. parapsilosis phagocytosis and reports intracellular survival mechanisms associated with this species.

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Section: Resultssupporting

confidence: 58%

Kinetic studies of Candida parapsilosis phagocytosis by macrophages and detection of intracellular survival mechanisms

Tóth

Tóth²,

Papp

et al. 2014

Front. Microbiol.

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“…Such analyses of the influence of geometric apposition of irregular-shaped pathogens encountered by host cells could not be conducted without live imaging. Recent work using similar methodology studying the interaction of the important plant and opportunistic human pathogen Fusarium oxysporum with macrophages revealed remarkable similarities with the findings described for C. albicans above [ 26 ]. The study shows that murine macrophages efficiently migrate towards and internalise F. oxysporum germlings.…”

Section: The Innate Immune Response To Fungal Infection In Vitromentioning

confidence: 52%

Novel insights into host-fungal pathogen interactions derived from live-cell imaging

2014

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The theoretical physicist and Nobel laureate Richard Feynman outlined in his 1959 lecture, “There’s plenty of room at the bottom”, the enormous possibility of producing and visualising things at smaller scales. The advent of advanced scanning and transmission electron microscopy and high-resolution microscopy has begun to open the door to visualise host-pathogen interactions at smaller scales, and spinning disc confocal and two-photon microscopy has improved our ability to study these events in real time in three dimensions. The aim of this review is to illustrate some of the advances in understanding host-fungal interactions that have been made in recent years in particular those relating to the interactions of live fungal pathogens with phagocytes. Dynamic imaging of host-pathogen interactions has recently revealed novel detail and unsuspected mechanistic insights, facilitating the dissection of the phagocytic process into its component parts. Here, we will highlight advances in our knowledge of host-fungal pathogen interactions, including the specific effects of fungal cell viability, cell wall composition and morphogenesis on the phagocytic process and try to define the relative contributions of neutrophils and macrophages to the clearance of fungal pathogens in vitro and the infected host.

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“…Algorithms for the automated image analysis of confrontation assays, especially in the context of fungal pathogens interacting with immune cells, have been developed before (Mech et al, 2011 , 2014 ; Kraibooj et al, 2014 ; Schäfer et al, 2014 ). The most important progress of the novel algorithm presented here concerns the successful segmentation of macrophages.…”

Section: Discussionmentioning

confidence: 99%

“…Hence, the staining protocol enabled us to conveniently work on single layers for object segmentation and to ultimately combine layers in the classification of objects and in the analysis of their spatial colocalizations. While some progress has been made in previous developments of algorithms for the automated image analysis of this type of experiments (Mech et al, 2011 , 2014 ; Kraibooj et al, 2014 ; Schäfer et al, 2014 ), we are here presenting a novel algorithm that differs from previous approaches with regard to the crucial step of segmenting all different types of cells in the assay. This was not achieved in previous approaches regarding the segmentation of macrophages, which is complicated by the occasionally interrupted staining of their cell surface.…”

Section: Introductionmentioning

confidence: 99%

Automated quantification of the phagocytosis of Aspergillus fumigatus conidia by a novel image analysis algorithm

et al. 2015

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Studying the pathobiology of the fungus Aspergillus fumigatus has gained a lot of attention in recent years. This is due to the fact that this fungus is a human pathogen that can cause severe diseases, like invasive pulmonary aspergillosis in immunocompromised patients. Because alveolar macrophages belong to the first line of defense against the fungus, here, we conduct an image-based study on the host-pathogen interaction between murine alveolar macrophages and A. fumigatus. This is achieved by an automated image analysis approach that uses a combination of thresholding, watershed segmentation and feature-based object classification. In contrast to previous approaches, our algorithm allows for the segmentation of individual macrophages in the images and this enables us to compute the distribution of phagocytosed and macrophage-adherent conidia over all macrophages. The novel automated image-based analysis provides access to all cell-cell interactions in the assay and thereby represents a framework that enables comprehensive computation of diverse characteristic parameters and comparative investigation for different strains. We here apply automated image analysis to confocal laser scanning microscopy images of the two wild-type strains ATCC 46645 and CEA10 of A. fumigatus and investigate the ability of macrophages to phagocytose the respective conidia. It is found that the CEA10 strain triggers a stronger response of the macrophages as revealed by a higher phagocytosis ratio and a larger portion of the macrophages being active in the phagocytosis process.

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Hyphal Growth of Phagocytosed Fusarium oxysporum Causes Cell Lysis and Death of Murine Macrophages

Cited by 10 publications

References 37 publications

Kinetic studies of Candida parapsilosis phagocytosis by macrophages and detection of intracellular survival mechanisms

Kinetic studies of Candida parapsilosis phagocytosis by macrophages and detection of intracellular survival mechanisms

Novel insights into host-fungal pathogen interactions derived from live-cell imaging

Automated quantification of the phagocytosis of Aspergillus fumigatus conidia by a novel image analysis algorithm

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