Quantification of zinc intoxication of Candida glabrata after phagocytosis by primary macrophages
| Autor: | Michael Riedelberger, Irina Tsymala, Sabrina Jenull, Philipp Penninger, Karl Kuchler, Hossein Arzani |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
inorganic chemicals
Phagocytosis Immunology chemistry.chemical_element Candida glabrata Zinc General Biochemistry Genetics and Molecular Biology Microbiology Cell Line chemistry.chemical_compound Mice Protocol Animals Humans Propidium iodide lcsh:Science (General) Fluorescent Dyes Zinc intoxication Innate immune system General Immunology and Microbiology biology General Neuroscience Macrophages Candidiasis Cell-based assays biology.organism_classification Bioavailability enzymes and coenzymes (carbohydrates) chemistry biological sciences health occupations bacteria Flow cytometry/mass cytometry Intracellular lcsh:Q1-390 |
| Zdroj: | STAR Protocols STAR Protocols, Vol 2, Iss 1, Pp 100352-(2021) |
| ISSN: | 2666-1667 |
| Popis: | 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). Graphical Abstract Highlights • A host-pathogen interaction system of primary macrophages and fungal pathogens • Flow cytometric assay to quantify Zn2+ intoxication of fungal pathogens by macrophages • Fungal viability depends on intra-phagosomal Zn2+ changes during immune responses • Zn2+ levels in reisolated pathogens correlate with fungal killing by macrophages 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. |
| Databáze: | OpenAIRE |
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