Killing of Cryptococcus neoformans by Staphylococcus aureus: The role of cryptococcal capsular polysaccharide in the fungal-bacteria interaction

Abstract

Microbes compete for the environmental niche which is their host. To investigate the effects of a pathogenic bacterium on invasion and colonization by a pathogenic yeast, Cryptococcus neoformans was co-cultured with Staphylococcus aureus. We found that the number of colony forming units of C. neoformans was decreased by Staphylococcus aureus. In contrast, the viability of Candida albicans was not affected. Under the microscope, wild-type C. neoformans cells were shown to be surrounded by S. aureus, while cells of a capsuleless mutant of C. neoformans were not. C. neoformans was not killed when a membrane separated it from S. aureus in co-culture. Killing was confirmed by staining with cyanoditolyl tetrazolium chloride: S. aureus stained red, indicating viability, while C. neoformans did not stain, indicating lethality. The in situ terminal deoxynucleotidyl transferase-mediated dUTR nick end labeling (TUNEL) assay indicated cell death with fragmentation of DNA of C. neoformans. Capsular polysaccharide from C. neoformans inhibited the killing. Treatment of the crude polysaccharide with protease increased the inhibition. The protective activity resided in the glucuronoxylomannan (GXM) fraction, although the concentration required for the inhibition was high. These results suggest that S. aureus kills C. neoformans by a process that involves attachment to the cryptococcal capsule. © 2004 ISHAM.