Role of host glycosphingolipids on Paracoccidioides brasiliensis adhesion.
Binding of yeast forms to human lung fibroblast cultures was analyzed, aiming to better understand the initial steps of Paracoccidioides brasiliensis infection in humans. A significant P. brasiliensis adhesion was observed either to fibroblasts or to their Triton X-100 insoluble fraction, which contains extracellular matrix and membrane microdomains enriched in glycosphingolipids. Since human lung fibroblasts express at cell-surface gangliosides, such as GM1, GM2, and GM3, the role of these glycosphingolipids on P. brasiliensis adhesion was analyzed by different procedures. Anti-GM3 monoclonal antibody or cholera toxin subunit B (which binds specifically to GM1) reduced significantly fungal adhesion to fibroblast cells, by 35% and 33%, respectively. Direct binding of GM1 to yeast forms of P. brasiliensis was confirmed using cholera toxin subunit B conjugated to AlexaFluor(®)488. It was also demonstrated that P. brasiliensis binds to polystyrene plates coated with galactosylceramide, lactosylceramide, trihexosylceramide, GD3, GM1, GM3, and GD1a, suggesting that glycosphingolipids presenting residues of beta-galactose or neuraminic acid at non-reducing end may act as adhesion molecules for P. brasiliensis. Conversely, no binding was detected when plates were adsorbed with glycosphingolipids that contain terminal residue of beta-N-acetylgalactosamine, such as globoside (Gb4), GM2, and asialo-GM2. In human fibroblast (WI-38 cells), GM3 and GM1 are associated with membrane rafts, which remain insoluble after treatment with Triton X-100 at 4°C. Taken together, these results strongly suggest that lung fibroblast gangliosides, GM3 and GM1, are involved in binding and/or infection by P. brasiliensis.
Department of Biochemistry, Universidade Federal de São Paulo, Rua Botucatu 862, São Paulo, SP, 04023-900, Brazil.
Cristina Y
CY
Ywazaki
Paloma K
PK
Maza
Erika
E
Suzuki
Helio K
HK
Takahashi
Anita H
AH
Straus
D002448:Cell Adhesion; D002460:Cell Line; D006028:Glycosphingolipids / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D008168:Lung / Q000378:metabolism / Q000382:microbiology; D010228:Paracoccidioides / Q000235:genetics / Q000502:physiology*; D010229:Paracoccidioidomycosis / Q000378:metabolism* / Q000382:microbiology*
D006028:Glycosphingolipids