Role of glucuronoxylomannan and steryl glucosides in protecting against cryptococcosis

The development of vaccines for fungal diseases, including cryptococcosis, is an emergent line of research and development. In previous studies, we showed that a <i>Cryptococcus</i> mutant lacking the <i>SGL1</i> gene (<i>∆sgl1</i>) accumulates certain glycolipids called steryl glucosides (SGs) on the fungal capsule, promoting an effective immunostimulation that totally protects the host from a secondary cryptococcal infection. However, this protection is lost when the cryptococcal capsule is absent in the <i>∆sgl1</i> background. The cryptococcal capsule is mainly composed of glucuronoxylomannan (GXM), a polysaccharide microfiber consisting of glucuronic acid, xylose, and mannose linked by glycosidic bonds forming specific triads. In this study, we engineered cells to lack each of the GXM components and tested the effect of these deletions on protection under the condition of SG accumulation. We found that glucuronic acid and xylose are required for protection, and their absence abrogates the production of IFNγ and IL-17A by γδ T cells, which are necessary stimulants for the protective phenotype of the <i>∆sgl1</i>. We analyzed the structure of the GXM microfibers and found that although the deletion of <i>SGL1</i> only slightly affects the size and distribution of these microfibers, it significantly changes the ratio of mannose to other components. In conclusion, this study identifies the structural modifications that the deletion of <i>SGL1</i> and the consequent accumulation of SGs impart to the GXM structure of <i>C. neoformans</i>. This provides significant insights into the protective mechanisms mediated by SG accumulation on the capsule, with important implications for the future development of an efficacious cryptococcal vaccine.IMPORTANCE<i>Cryptococcus neoformans</i> is an encapsulated fungus that causes invasive fungal infections with high morbidity and mortality in susceptible patients. With increasing drug resistance and high toxicity of current antifungal drugs, there is a need for alternative therapeutic strategies, such as a cryptococcal vaccine. In this study, we identify the necessary capsular components and their structural organization required for a cryptococcal vaccine to protect the host against challenge with a virulent strain. These capsular components are glucuronic acid, xylose, and mannose, and they work together with certain glycolipids called steryl glucosides (SGs) to stimulate host immunity. Interestingly, SGs on the capsule may favor the formation of small capsular microfibers organized in specific mannose triads. Thus, the results of this paper are important because they identify a mechanism by which SGs affect the structure of the cryptococcal capsule, with important implications for the future development of a cryptococcal vaccine using capsular components and SGs.