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J. Biol. Chem., Vol. 281, Issue 32, 22614-22623, August 11, 2006

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An -Glucan of Pseudallescheria boydii Is Involved in Fungal Phagocytosis and Toll-like Receptor Activation^*

Vera Carolina B. Bittencourt¹, Rodrigo T. Figueiredo¹, Rosana B. da Silva, Diego S. Mourão-Sá, Patricia L. Fernandez, Guilherme L. Sassaki^¶, Barbara Mulloy^||, Marcelo T. Bozza², and Eliana Barreto-Bergter³

From the Departamento de Microbiologia Geral and Departamento de Imunologia, Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Avenida Brigadeiro Trompowsky s/n, CCS Bloco I, 21941-590 Rio de Janeiro, Brazil, the ^¶Departamento de Bioquímica, Universidade Federal do Paraná, 81531-990 Curitiba, Paraná, Brazil, and the ^||National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom

The host response to fungi is in part dependent on activation of evolutionarily conserved receptors, including toll-like receptors and phagocytic receptors. However, the molecular nature of fungal ligands responsible for this activation is largely unknown. Herein, we describe the isolation and structural characterization of an -glucan from Pseudallescheria boydii cell wall and evaluate its role in the induction of innate immune response. These analyses indicate that -glucan of P. boydii is a glycogen-like polysaccharide consisting of linear 4-linked -D-Glcp residues substituted at position 6 with -D-Glcp branches. Soluble -glucan, but not -glucan, led to a dose-dependent inhibition of conidia phagocytosis. Furthermore, a significant decrease in the phagocytic index occurred when -glucan from conidial surface was removed by enzymatic treatment with -amyloglucosidase, thus indicating an essential role of -glucan in P. boydii internalization by macrophages. -Glucan stimulates the secretion of inflammatory cytokines by macrophages and dendritic cells; again this effect is abolished by treatment with -amyloglucosidase. Finally, -glucan induces cytokine secretion by cells of the innate immune system in a mechanism involving toll-like receptor 2, CD14, and MyD88. These results might have relevance in the context of infections with P. boydii and other fungi, and -glucan could be a target for intervention during fungal infections.

Received for publication, October 20, 2005 , and in revised form, June 9, 2006.

^* The work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico, Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior, Fundação de Amparo a Pesquisa do Estado do Rio de Janeiro, and Programa de Núcleos de Excelência, and the Universidade Federal do Rio de Janeiro (UFRJ). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally to this work.

² To whom correspondence may be addressed. Tel.: 55-21-8729-5029; Fax: 55-21-2560-8344; E-mail: mbozza{at}micro.ufrj.br. ³ To whom correspondence may be addressed. Tel.: 55-21-2562-6741; Fax: 55-21-2560-8344; E-mail: eliana.bergter{at}micro.ufrj.br.

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