Inhibition of PbGP43 Expression May Suggest that gp43 is a Virulence Factor in Paracoccidioides brasiliensis

dc.contributor.authorTorres, Isaura
dc.contributor.authorHernandez, Orville
dc.contributor.authorTamayo, Diana
dc.contributor.authorMunoz, Jose F.
dc.contributor.authorLeitão Junior, Natanael P. [UNIFESP]
dc.contributor.authorGarcia, Ana M.
dc.contributor.authorRestrepo, Angela
dc.contributor.authorPuccia, Rosana [UNIFESP]
dc.contributor.authorMcEwen, Juan G.
dc.contributor.institutionUniv Antioquia
dc.contributor.institutionInst Univ Colegio Mayor Antioquia
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.description.abstractGlycoprotein gp43 is an immunodominant diagnostic antigen for paracoccidioidomycosis caused by Paracoccidioides brasiliensis. It is abundantly secreted in isolates such as Pb339. It is structurally related to beta-1,3-exoglucanases, however inactive. Its function in fungal biology is unknown, but it elicits humoral, innate and protective cellular immune responses; it binds to extracellular matrix-associated proteins. in this study we applied an antisense RNA (aRNA) technology and Agrobacterium tumefaciens-mediated transformation to generate mitotically stable PbGP43 mutants (PbGP43 aRNA) derived from wild type Pb339 to study its role in P. brasiliensis biology and during infection. Control PbEV was transformed with empty vector. Growth curve, cell vitality and morphology of PbGP43 aRNA mutants were indistinguishable from those of controls. PbGP43 expression was reduced 80-85% in mutants 1 and 2, as determined by real time PCR, correlating with a massive decrease in gp43 expression. This was shown by immunoblotting of culture supernatants revealed with anti-gp43 mouse monoclonal and rabbit polyclonal antibodies, and also by affinity-ligand assays of extracellular molecules with laminin and fibronectin. in vitro, there was significantly increased TNF-alpha production and reduced yeast recovery when PbGP43 aRNA1 was exposed to IFN-gamma-stimulated macrophages, suggesting reduced binding/uptake and/or increased killing. in vivo, fungal burden in lungs of BALB/c mice infected with silenced mutant was negligible and associated with decreased lung I Lambda-10 and IL-6. Therefore, our results correlated low gp43 expression with lower pathogenicity in mice, but that will be definitely proven when PbGP43 knockouts become available. This is the first study of gp43 using genetically modified P. brasiliensis.en
dc.description.affiliationCIB, Unidad Biol Celular & Mol, Medellin, Colombia
dc.description.affiliationUniv Antioquia, Fac Med, Medellin, Colombia
dc.description.affiliationUniv Antioquia, Inst Biol, Medellin, Colombia
dc.description.affiliationInst Univ Colegio Mayor Antioquia, Fac Ciencias Salud, Medellin, Colombia
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipColciencias project
dc.description.sponsorshipCorporacion para Investigaciones Biologicas
dc.description.sponsorshipUniversidad de Antioquia Estrategia de Sostenibilidad
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipIDColciencias project: 221340820447
dc.identifier.citationPlos One. San Francisco: Public Library Science, v. 8, n. 7, 9 p., 2013.
dc.publisherPublic Library Science
dc.relation.ispartofPlos One
dc.rightsAcesso aberto
dc.titleInhibition of PbGP43 Expression May Suggest that gp43 is a Virulence Factor in Paracoccidioides brasiliensisen
Pacote Original
Agora exibindo 1 - 1 de 1
Imagem de Miniatura
954.19 KB
Adobe Portable Document Format