Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity

Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity

Autor Tanaka, Améria Kaori Autor UNIFESP Google Scholar
Gorin, Philip Albert James Google Scholar
Takahashi, Helio Kiyoshi Autor UNIFESP Google Scholar
Straus, Anita Hilda Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Universidade Federal do Paraná Departamento de Bioquímica
Resumo The role of glycosphingolipids (GSLs) present in amastigote forms of Leishmania (Leishmania) amazonensis during infection of macrophages was analyzed, with particular emphasis on GSLs presenting the terminal Galpß1-3Galpa disaccharide. Macrophage invasion by L. (L.) amazonensis amastigotes was reduced by 37% when the disaccharide Galpß1-3Galp (1 mM) was added to the culture medium. The putative macrophage receptor/lectin for ß-Gal-globotriaosylceramide (Galpß1-3Galpa1-4Galpß1-4Glc pß1-1Cer) and other structurally related GSLs from L. (L.) amazonensis amastigotes were analyzed by micelles and parasite binding assay to peritoneal macrophage proteins fractionated by SDS-PAGE under nonreducing conditions. Micelles containing purified amastigote GSLs or a suspention of L. (L.) amazonensis amastigotes fixed with 2% formaldehyde were incubated with nitrocellulose membrane containing the macrophage proteins transferred by Western blotting. Binding of micelles containing purified GSLs from amastigote forms or fixed L. (L.) amazonensis amastigotes to nitrocellulose membrane was probed using monoclonal antibody ST-3, which recognizes the glycoepitope Galpß1-3Galpa1-R present either in the micelle preparation or on the amastigote surface. Macrophage protein with molecular mass ~30 kDa bound the amastigote GSL and appeared to be a doublet on electrophoresis. The specificity of this interaction was confirmed using fixed L. (L.) chagasi amastigotes, which do not express GSLs such as ß-Galp-globotriaosylceramides, and which do not bind to 30-kDa protein.
Palavra-chave Leishmania (Leishmania) amazonensis
Amastigote
Glycosphingolipid
Macrophage binding protein
Idioma Inglês
Data de publicação 2007-06-01
Publicado em Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 40, n. 6, p. 799-806, 2007.
ISSN 0100-879X (Sherpa/Romeo, fator de impacto)
Publicador Associação Brasileira de Divulgação Científica
Extensão 799-806
Fonte http://dx.doi.org/10.1590/S0100-879X2006005000106
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000248409900008
SciELO S0100-879X2007000600008 (estatísticas na SciELO)
Endereço permanente http://repositorio.unifesp.br/handle/11600/3777

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