Abstract
Mammalian infection by dimorphic fungi occurs through the inhalation of asexual spores (conidia), which are phagocytosed by host pulmonary alveolar macrophages of the innate immune system. Once phagocytosed, fungal conidia germinate into the pathogenic cell type; unicellular yeast cells which divide by fission (Vanittanakom et al., 2006; Boyce et al., 2011). To investigate if mutation of a particular fungal gene affects macrophage phagocytosis or the production of yeast cells, a murine macrophage cell culture assay can be utilized. This protocol was developed for Penicillium marneffei but is applicable to most dimorphic fungi.
Materials and Reagents
Equipment
Procedure
Note: All steps performed in the Biological Safety Cabinet. Use sterile materials and reagents and aseptic techniques. All incubation steps are at 37 °C, 5% CO2 in a cell culture incubator unless indicated. Day 1
Recipes
Acknowledgments
This work was funded by grants from the Australian Research Council, National Health and Medical Research Council of Australia and the Howard Hughes Medical Institute.
References
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