Landgraf Taise N, Costa Marcelo V, Oliveira Aline F, Ribeiro Wander C, Panunto-Castelo Ademilson, Fernandes Fabrício F
Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São PauloRibeirão Preto, Brazil.
Department of Biology, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São PauloRibeirão Preto, Brazil.
Front Microbiol. 2017 Sep 20;8:1803. doi: 10.3389/fmicb.2017.01803. eCollection 2017.
and are fungi causing paracoccidioidomycosis (PCM), an autochthonous systemic mycosis found in Latin America. These microorganisms contain a multitude of molecules that may be associated with the complex interaction of the fungus with the host. Here, we identify the enzyme dihydrolipoyl dehydrogenase (DLD) as an exoantigen from (Pb18_Dld) by mass spectrometry. Interestingly, the DLD gene expression in yeast form showed higher expression levels than those in mycelial form and transitional phases. Pb18_Dld gene was cloned, and the recombinant protein (rPb18_Dld) was expressed and purified for subsequent studies and production of antibodies. Immunogold labeling and transmission electron microscopy revealed that the Pb18_Dld is also localized in mitochondria and cytoplasm of . Moreover, when macrophages were stimulated with rPb18Dld, there was an increase in the phagocytic and microbicidal activity of these cells, as compared with non-stimulated cells. These findings suggest that Pb18_Dld can be involved in the pathogen-host interaction, opening possibilities for studies of this protein in PCM.
和是引起副球孢子菌病(PCM)的真菌,PCM是一种在拉丁美洲发现的地方性系统性真菌病。这些微生物含有多种分子,这些分子可能与真菌与宿主的复杂相互作用有关。在这里,我们通过质谱法将二氢硫辛酰胺脱氢酶(DLD)鉴定为**(Pb18_Dld)的一种外抗原。有趣的是,酵母形式的DLD基因表达水平高于菌丝体形式和过渡阶段。克隆了Pb18_Dld基因,并表达和纯化了重组蛋白(rPb18_Dld),用于后续研究和抗体生产。免疫金标记和透射电子显微镜显示,Pb18_Dld也定位于的线粒体和细胞质中**。此外,用rPb18Dld刺激巨噬细胞时,与未刺激的细胞相比,这些细胞的吞噬和杀菌活性有所增加。这些发现表明,Pb18_Dld可能参与病原体与宿主的相互作用,为在PCM中研究这种蛋白质开辟了可能性。