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Role of Host Glycosphingolipids on Paracoccidioides brasiliensis Adhesion

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Abstract

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.

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Acknowledgments

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Pesquisa (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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Authors and Affiliations

  1. Department of Biochemistry, Universidade Federal de São Paulo, Rua Botucatu 862, São Paulo, SP, 04023-900, Brazil

    Cristina Y. Ywazaki, Paloma K. Maza, Erika Suzuki, Helio K. Takahashi & Anita H. Straus

  2. Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo, Rua Botucatu 862, São Paulo, SP, 04023-900, Brazil

    Paloma K. Maza & Erika Suzuki

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  1. Cristina Y. Ywazaki
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  2. Paloma K. Maza
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  5. Anita H. Straus
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Correspondence to Anita H. Straus.

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Ywazaki, C.Y., Maza, P.K., Suzuki, E. et al. Role of Host Glycosphingolipids on Paracoccidioides brasiliensis Adhesion. Mycopathologia 171, 325–332 (2011). https://doi.org/10.1007/s11046-010-9376-4

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  • DOI: https://doi.org/10.1007/s11046-010-9376-4

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