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

Abstract

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: Cell-based assays; Flow cytometry/mass cytometry; Immunology.