Quantification of zinc intoxication of Candida glabrata after phagocytosis by primary macrophages

Abstract

Summary: Zinc (Zn2+) is a trace element, playing pivotal roles during host-pathogen interactions. Macrophages can sequester Zn2+ and restrict bioavailability or increase phagolysosomal Zn2+ to kill pathogens. This method quantifies Zn2+-mediated clearance of the human fungal pathogen C. glabrata after phagocytosis by innate immune cells. Double staining with propidium iodide and a zinc-specific fluorescence dye allows for discrimination of live versus dead pathogens inside phagolysosomes. Moreover, elevated phagolysosomal Zn2+ decreases fungal viability as a function of intracellular Zn2+ concentrations in macrophages.For complete details on the use and execution of this protocol, please refer to Riedelberger et al. (2020).

Keywords

• Flow cytometry/mass cytometry
• Cell-based assays
• Immunology