Divalent metal cations potentiate the predatory capacity of amoeba for Cryptococcus neoformans

Abstract

Among the best studied interaction between soil phagocytic predators and a human pathogenic fungus is that of Acanthamoeba castellanii and Cryptococcus neoformans. The experimental conditions used in amoeba-fungal confrontation assays can have major effects on whether the fungus or the protozoan is ascendant in the interaction. In the presence of Mg2+ and Ca2+ in PBS, C. neoformans was consistently killed when incubated with A. castellanii. A. castellanii survived better in the presence of Mg2+ and Ca2+, even when incubated with C. neoformans. In the absence of Mg2+ and Ca2+, C. neoformans survived when incubated with A. castellanii, and the percentage of dead amoeba was higher than when incubated without yeast cells. These results show that the presence of Mg2+ and Ca2+ can make a decisive contribution toward tilting the outcome of the interaction in favor of amoeba. Of the two metals Mg2+ had a stronger effect than Ca2+. Cations enhanced A. castellanii activity against C. neoformans through enhanced phagocytosis, which is the major mechanism for amoeba to kill fungal cells. We found no evidence that amoeba uses extracellular killing mechanisms in their interactions with C. neoformans. In summary, the presence of Mg2+ and Ca2+ enhanced cell adhesion on surface and motility of amoeba, thus increasing the chance for contact of C. neoformans and the frequency of phagocytosis. Our findings imply that divalent cation concentration in soils could be an important variable for whether amoeba can control C. neoformans in the environment.\n\nImportanceGrazing of soil organisms by phagocytic predators such as amoeba is thought to select for traits that allow some of them to acquire the capacity for virulence in animals. Consequently, knowledge about the interactions between amoeba and soil microbes, such as pathogenic fungi, is important for understanding how virulence can emerge. We show that the interaction between amoeba and the pathogenic fungus C. neoformans is influenced by the presence of magnesium and calcium in the assay, which potentiate amoeba. The results may also have practical applications since enriching soils with divalent cations may reduce C. neoformans numbers in contaminated soils.