Abstract
The polysaccharide capsule of fungal pathogen Cryptococcus neoformans is a critical virulence factor that has historically evaded characterization. Polysaccharides remain attached to the cell as capsular polysaccharide (CPS) or are shed into the surroundings in the form of exopolysaccharide (EPS). While a great deal of study has been done examining the properties of EPS, far less is known about CPS. In this work, we detail the development of sonication, French Press, and glucanex digestion to both remove the capsule and produce ‘capsule ghosts.’ The ability of sonication to strip cells of capsular material while maintaining survival allowed us to explore of capsule regrowth dynamics, immune modulation, as well as NMR characterization of removed material. Development of these new methodologies will allow for exciting future work exploring cell wall capsule dynamics and improving the characterization of CPS.
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