title: Extracellular vesicles from Paracoccidioides pathogenic species transport polysaccharide and expose ligands for DC-SIGN receptors.
abstract: Extracellular vesicles (EVs) mediate non-conventional transport of molecules across the fungal cell wall. We aimed at describing the carbohydrate composition and surface carbohydrate epitopes of EVs isolated from the pathogenic fungi Paracoccidioides brasiliensis and P. lutzii using standard procedures. Total EV carbohydrates were ethanol-precipitated from preparations depleted of lipids and proteins, then analyzed by chemical degradation, gas chromatography-mass spectrometry, nuclear magnetic resonance and size-exclusion chromatography. EV glycosyl residues of Glc, Man, and Gal comprised most probably two major components: a high molecular mass 4,6-α-glucan and a galactofuranosylmannan, possibly an oligomer, bearing a 2-α-Manp main chain linked to β-Galf (1,3) and α-Manp (1,6) end units. The results also suggested the presence of small amounts of a (1→6)-Manp polymer, (1→3)-glucan and (1→6)-glucan. Glycan microarrays allowed identification of EV surface lectin(s), while plant lectin microarray profiling revealed terminal Man and GlcNAc residues exposed at the EVs surface. Mammalian lectin microarray profiling showed that DC-SIGN receptors recognized surface carbohydrate in Paracoccidioides EVs. Our results suggest that oligosaccharides, cytoplasmic storage, and cell wall polysaccharides can be exported in fungal EVs, which also expose surface PAMPs and lectins. The role of these newly identified components in the interaction with the host remains to be unraveled.
authors.affiliation: Escola Paulista de Medicina-Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Roberta
authors.initials: R
authors.lastname: Peres da Silva
authors.affiliation: Analytical Services &Training Laboratory Complex Carbohydrate Research Center, The University of Georgia.
authors.forename: Christian
authors.initials: C
authors.lastname: Heiss
authors.affiliation: Analytical Services &Training Laboratory Complex Carbohydrate Research Center, The University of Georgia.
authors.forename: Ian
authors.initials: I
authors.lastname: Black
authors.affiliation: Analytical Services &Training Laboratory Complex Carbohydrate Research Center, The University of Georgia.
authors.forename: Parastoo
authors.initials: P
authors.lastname: Azadi
authors.affiliation: Glycoscience Group, National Centre for Biomedical Engineering Science and.
authors.forename: Jared Q
authors.initials: JQ
authors.lastname: Gerlach
authors.affiliation: Escola Paulista de Medicina-Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Luiz R
authors.initials: LR
authors.lastname: Travassos
authors.affiliation: Glycoscience Group, National Centre for Biomedical Engineering Science and.
authors.forename: Lokesh
authors.initials: L
authors.lastname: Joshi
authors.affiliation: Glycoscience Group, National Centre for Biomedical Engineering Science and.|Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
authors.forename: Michelle
authors.initials: M
authors.lastname: Kilcoyne
authors.affiliation: Escola Paulista de Medicina-Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Rosana
authors.initials: R
authors.lastname: Puccia