Purification and Immunomodulatory activity of the capsular polysaccharides mating types a and a from Cryptococcus neoformans var. neoformans
Villena, SN1,2; Pinheiro RO2; Pinheiro, C2; Luna, TGS2; Nunes MP2; Previato, JO1; Mendonça-Previato L1; DosReis GA2 & Freire de Lima2, CG.
1Laboratório de Glicobiologia, 2Laboratório de Biologia Imunitária, Instituto de Biofísica Carlos Chagas Filho, CCS, UFRJ, Ilha do Fundão, 21944-970, Rio de Janeiro, RJ.
Cryptococcus neoformans is an opportunistic yeast that is surrounded by a polysaccharide capsule, which is the most prominent virulence factor from this fungus. The capsule is principally constituted by glucuronoxylomannan (GXM) representing 88% of total capsule weight, galactoxylomannan (GalXM) representing the other 12%, together with some mannoproteins (MP). The C. neoformans possesses a bipolar system with two mating types, MATa and MATα. Genetic analysis has demonstrated (1:1) segregation of mating types in laboratory crosses. However, clinical samples and environmental isolates are predominantly of the MATα, ranging from (30:1 – α:a) and (40:1 – α:a) respectively, a fact that could explain the major virulence of this mating type. Previous results in our work have demonstrated that both polysaccharides (GXM and GalXM) induced an intensive vacuolization and a reduction in cellular viability in RAW 264.7 murine macrophages lineage, especially in GalXM in both mating types. This reduction was due to cell apoptosis, being recovered when cells were treated with a neutralizing anti-FasL antibody, suggesting an involvement of Fas-FasL pathway. Also, we observed that GalXM in both mating types induced a more pronounced nitric oxide (NO) production than GXM. However, TGF- b production was observed only in cell cultures treated with GXMα, 24h after stimulation. Apoptosis induced in RAW cell line was a mechanism dependent of the Fas-L expression on the cell surface, and this mechanism was more induced when macrophages were stimulated by GalXM. On the other hand, GXM induces apoptosis and TGF- b increases in cell cultures. These results suggest that both capsular polysaccharides from Cryptococcus neoformans display immunomodulatory properties on macrophage biology.
FAPERJ, CNPq (PRONEX) & HHMI
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