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Sarah Ghamrawi - One of the best experts on this subject based on the ideXlab platform.
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RESEARCH ARTICLE A Multifaceted Study of Scedosporium boydii Cell Wall Changes during Germination and Identification of GPI-Anchored Proteins
2016Co-Authors: Sarah Ghamrawi, Agnès Marot, Ine Gastebois, Agata Zykwinska, Guillaume Mabilleau, Stéphane CuenotAbstract:Scedosporium boydii is a pathogenic filamentous fungus that causes a wide range of human infections, notably respiratory infections in patients with cystic fibrosis. The development of new therapeutic strategies targeting S. boydii necessitates a better understanding of the phys-iology of this fungus and the identification of newmolecular targets. In this work, we studied the conidium-to-germ tube transition using a variety of techniques including scanning and transmission electron microscopy, atomic forcemicroscopy, two-phase partitioning, micro-electrophoresis and cationized ferritin labeling, chemical force spectroscopy, lectin labeling, and nanoLC-MS/MS for cell wall GPI-anchored protein analysis. We demonstrated that the cell wall undergoes structural changes with germination accompanied with a lower hydropho-bicity, electrostatic charge and binding capacity to cationized ferritin. Changes during germina-tion also included a higher accessibility of some cell wall polysaccharides to lectins and less CH3/CH3 interactions (hydrophobic adhesion forces mainly due to glycoproteins). We also ex-tracted and identified 20 GPI-anchored proteins from the cell wall of S. boydii, among which one was detected only in the conidial wall extract and 12 only in the mycelial wall extract. Th
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A Multifaceted Study of Scedosporium boydii Cell Wall Changes during Germination and Identification of GPI-Anchored Proteins
2015Co-Authors: Sarah Ghamrawi, Agnès Marot, Patrick Vandeputte, Amandine Gastebois, Agata Zykwinska, Guillaume Mabilleau, Stéphane Cuenot, Jean-philippe BoucharaAbstract:Scedosporium boydii is a pathogenic filamentous fungus that causes a wide range of human infections, notably respiratory infections in patients with cystic fibrosis. The development of new therapeutic strategies targeting S. boydii necessitates a better understanding of the physiology of this fungus and the identification of new molecular targets. In this work, we studied the conidium-to-germ tube transition using a variety of techniques including scanning and transmission electron microscopy, atomic force microscopy, two-phase partitioning, microelectrophoresis and cationized ferritin labeling, chemical force spectroscopy, lectin labeling, and nanoLC-MS/MS for cell wall GPI-anchored protein analysis. We demonstrated that the cell wall undergoes structural changes with germination accompanied with a lower hydrophobicity, electrostatic charge and binding capacity to cationized ferritin. Changes during germination also included a higher accessibility of some cell wall polysaccharides to lectins and less CH3/CH3 interactions (hydrophobic adhesion forces mainly due to glycoproteins). We also extracted and identified 20 GPI-anchored proteins from the cell wall of S. boydii, among which one was detected only in the conidial wall extract and 12 only in the mycelial wall extract. The identified sequences belonged to protein families involved in virulence in other fungi like Gelp/Gasp, Crhp, Bglp/Bgtp families and a superoxide dismutase. These results highlighted the cell wall remodeling during germination in S. boydii with the identification of a substantial number of cell wall GPI-anchored conidial or hyphal specific proteins, which provides a basis to investigate the role of these molecules in the host-pathogen interaction and fungal virulence.
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Modifications de la paroi au cours de la maturation et de la germination des conidies de Scedosporium boydii
2014Co-Authors: Sarah GhamrawiAbstract:Les espèces du complexe Scedosporium apiospermum sont des agents pathogènes émergents qui se situent au deuxième rang parmi les champignons filamenteux rencontrés au cours de la mucoviscidose. Ils sont omniprésents et particulièrement rencontrés dans les zones polluées. En dépit de leur importance clinique, nos connaissances sur leur biologie moléculaire et leur physiologie restent limitées. Chez les champignons, la paroi constitue un bouclier protecteur face à des conditions environnementales défavorables, et joue un rôle essentiel dans la pathogénicité. Ici, nous avons étudié les changements dynamiques de la paroi des conidies de S. boydii, l’une des deux espèces majeures de ce complexe avec S. apiospermum, avec pour objectif d'identifier des facteurs de virulence potentiels. En utilisant une large variété de techniques, allant de la microscopie électronique à balayage ou à transmission à l’analyse protéomique des protéines à ancre glycosylphosphatidylinositol (GPI) en passant par la microélectrophorèse et la partition de phase, la cytométrie en flux, la microscopie de force atomique, la résonance paramagnétique électronique, ou encore des techniques moléculaires, nous avons mis en évidence diverses modifications qui se produisent dans la paroi pendant la maturation et la germination des conidies de S. boydii et nous avons identifié la DHN-mélanine ainsi qu'un nombre important de protéines à ancre GPI. Enfin, nous avons fourni la première séquence complète du génome de S. apiospermum qui appuierait les différents domaines de la recherche sur ces champignons que ce soit pour l’étude des mécanismes pathogènes ou pour des applications biotechnologiques.
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modifications de la paroi au cours de la maturation et de la germination des conidies de Scedosporium boydii
2014Co-Authors: Sarah GhamrawiAbstract:Les especes du complexe Scedosporium apiospermum sont des agents pathogenes emergents qui se situent au deuxieme rang parmi les champignons filamenteux rencontres au cours de la mucoviscidose. Ils sont omnipresents et particulierement rencontres dans les zones polluees. En depit de leur importance clinique, nos connaissances sur leur biologie moleculaire et leur physiologie restent limitees. Chez les champignons, la paroi constitue un bouclier protecteur face a des conditions environnementales defavorables, et joue un role essentiel dans la pathogenicite. Ici, nous avons etudie les changements dynamiques de la paroi des conidies de S. boydii, l’une des deux especes majeures de ce complexe avec S. apiospermum, avec pour objectif d'identifier des facteurs de virulence potentiels. En utilisant une large variete de techniques, allant de la microscopie electronique a balayage ou a transmission a l’analyse proteomique des proteines a ancre glycosylphosphatidylinositol (GPI) en passant par la microelectrophorese et la partition de phase, la cytometrie en flux, la microscopie de force atomique, la resonance paramagnetique electronique, ou encore des techniques moleculaires, nous avons mis en evidence diverses modifications qui se produisent dans la paroi pendant la maturation et la germination des conidies de S. boydii et nous avons identifie la DHN-melanine ainsi qu'un nombre important de proteines a ancre GPI. Enfin, nous avons fourni la premiere sequence complete du genome de S. apiospermum qui appuierait les differents domaines de la recherche sur ces champignons que ce soit pour l’etude des mecanismes pathogenes ou pour des applications biotechnologiques.
Sara Mina - One of the best experts on this subject based on the ideXlab platform.
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Scedosporium boydii cata1 and sodc recombinant proteins new tools for serodiagnosis of Scedosporium infection of patients with cystic fibrosis
Diagnostic Microbiology and Infectious Disease, 2017Co-Authors: Sara Mina, Agnès Marot, Jean-philippe Bouchara, Cindy Staerck, Alphonse Calenda, Charlotte Godon, Maxime FleuryAbstract:Scedosporium species rank the second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF), after Aspergillus fumigatus. In CF, these fungi may cause various respiratory infections similar to those caused by A. fumigatus, including bronchitis and allergic broncho-pulmonary mycoses. Diagnosis of these infections relies on the detection of serum antibodies using crude antigenic extracts. However, many components of these extracts are common to Scedosporium and Aspergillus species, leading to cross-reactions. Here, 5 recombinant proteins from S. apiospermum or S. boydii were produced, and their value in serodiagnosis of Scedosporium infections was investigated by enzyme-linked immunosorbent assay. Two of them, corresponding to the Scedosporium catalase A1 or cytosolic Cu,Zn-superoxyde dismutase, allowed the detection of Scedosporium infection, and the differentiation with an Aspergillus infection. These recombinant proteins therefore may serve as a basis for the development of a standardized serological test.
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identification of Scedosporium boydii catalase a1 gene a reactive oxygen species detoxification factor highly expressed in response to oxidative stress and phagocytic cells
Fungal Biology, 2015Co-Authors: Sara Mina, Agnès Marot, Cindy Staerck, S Dalmeida, Yves Delneste, Alphonse Calenda, Julie TabiascoAbstract:Scedosporium boydii is an opportunistic filamentous fungus which may be responsible for a large variety of infections in both immunocompetent and immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF). Species of the S. apiospermum complex are able to chronically colonize the CF airways suggesting pathogenic mechanisms allowing persistence and growth of these fungi in the respiratory tract. Few putative virulence factors have been purified and characterized so far in the S. apiospermum complex including a cytosolic Cu,Zn-superoxide dismutase (SOD) and a monofunctional catalase (catalase A1). Upon microbial infection, host phagocytes release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by degradation of the hydrogen peroxide. Here, we identified the S. boydii catalase A1 gene (CATA1) and investigated its expression in response to the environmental conditions encountered in the CF airways and to the oxidative stress. Results showed that S. boydii CATA1 gene expression is not affected by hypoxia, hypercapnia or pH changes. In contrast, CATA1 gene was overexpressed in response to a chemically induced oxidative stress with a relative gene expression 37-fold higher in the presence of 250 μM H(2)O(2), 20-fold higher with 250 μM menadione and 5-fold higher with 2 mM paraquat. Moreover, S. boydii CATA1 gene expression progressively increased upon exposure to activated THP-1-derived macrophages, reaching a maximum after 12 h (26 fold). Activated HL60-derived neutrophils and activated human peripheral blood neutrophils more rapidly induced S. boydii CATA1 gene overexpression, a maximum gene expression level being reached at 75 min (17 fold) and 60 min (15 fold), respectively. In contrast expression of the gene encoding the Cu,Zn-SOD (SODC gene) was not affected by H(2)O(2), menadione, paraquat or in co-culture with phagocytic cells. These results suggest that S. boydii CATA1 gene is highly stimulated by the oxidative burst response whereas SODC gene is constitutively expressed.
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Purification and characterization of a mycelial catalase from Scedosporium boydii, a useful tool for specific antibody detection in patients with cystic fibrosis
Clinical and Vaccine Immunology, 2015Co-Authors: Sara Mina, Agnès Marot, Bernard Cimon, Maxime Fleury, Gérald Larcher, Jean-philippe Bouchara, Raymond RobertAbstract:Scedosporium boydii is an opportunistic filamentous fungus which may be responsible for a wide variety of infections in immunocompetent as well as immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF) and that may lead to allergic broncho-pulmonary mycoses, sensitization or respiratory infections. Upon microbial infection, host phagocytic cells release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by detoxification of the hydrogen peroxide. Here, we investigated the catalase equipment of S. boydii, one of the major pathogenic species in the S. apiospermum species complex. Three catalases were identified and the mycelial catalase A1 was purified to homogeneity by a three-step chromatographic process. This enzyme is a monofunctional tetrameric protein, of 460 kDa, consisting of four 82-kDa glycosylated subunits. The potential interest of this enzyme in serodiagnosis of S. apiospermum infections was then investigated by ELISA, using 64 sera from CF patients. Whatever the species involved in the S. apiospermum complex, sera from infected patients were clearly differentiated from sera from patients with an Aspergillus fumigatus infection, or from CF patients without clinical and biological signs of a fungal infection and without any fungus recovered from sputum samples. These results suggest that catalase A1 is a good candidate for the development of an immunoassay for serodiagnosis of infetions caused by the S. apiospermum complex in patients with CF.
Maxime Fleury - One of the best experts on this subject based on the ideXlab platform.
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Scedosporium boydii cata1 and sodc recombinant proteins new tools for serodiagnosis of Scedosporium infection of patients with cystic fibrosis
Diagnostic Microbiology and Infectious Disease, 2017Co-Authors: Sara Mina, Agnès Marot, Jean-philippe Bouchara, Cindy Staerck, Alphonse Calenda, Charlotte Godon, Maxime FleuryAbstract:Scedosporium species rank the second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF), after Aspergillus fumigatus. In CF, these fungi may cause various respiratory infections similar to those caused by A. fumigatus, including bronchitis and allergic broncho-pulmonary mycoses. Diagnosis of these infections relies on the detection of serum antibodies using crude antigenic extracts. However, many components of these extracts are common to Scedosporium and Aspergillus species, leading to cross-reactions. Here, 5 recombinant proteins from S. apiospermum or S. boydii were produced, and their value in serodiagnosis of Scedosporium infections was investigated by enzyme-linked immunosorbent assay. Two of them, corresponding to the Scedosporium catalase A1 or cytosolic Cu,Zn-superoxyde dismutase, allowed the detection of Scedosporium infection, and the differentiation with an Aspergillus infection. These recombinant proteins therefore may serve as a basis for the development of a standardized serological test.
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the secreted polyketide boydone a is responsible for the anti staphylococcus aureus activity of Scedosporium boydii
Fems Microbiology Letters, 2017Co-Authors: Cindy Staerck, Gérald Larcher, Jean-philippe Bouchara, Anne Landreau, Gaetan Herbette, Catherine Roullier, Samuel Bertrand, Benjamin Siegler, Maxime FleuryAbstract:Usually living as a soil saprophyte, the filamentous fungus Scedosporium boydii may also cause various infections in human. Particularly, it is one of the major causative agents of fungal colonization of the airways in patients with cystic fibrosis (CF). To compete with other microorganisms in the environment, fungi have evolved sophisticated strategies, including the production of secondary metabolites with antimicrobial activity that may also help them to establish successfully within the respiratory tract of receptive hosts. Here, the culture filtrate from a human pathogenic strain of S. boydii was investigated searching for an antibacterial activity, mainly against the major CF bacterial pathogens. A high antibacterial activity against Staphylococcus aureus, including methicillin-resistant strains of this species, was observed. Bio-guided fractionation and analysis of the active fractions by nuclear magnetic resonance or by high-performance liquid chromatography and high-resolution electrospray ionization-mass spectrometry allowed us to identify boydone A as responsible for this antibacterial activity. Together, these results suggest that this six-membered cyclic polyketide could be one of the virulence factors of the fungus. Genes involved in the synthesis of this secreted metabolite are currently being identified in order to confirm the role of this polyketide in pathogenesis.
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Purification and characterization of a mycelial catalase from Scedosporium boydii, a useful tool for specific antibody detection in patients with cystic fibrosis
Clinical and Vaccine Immunology, 2015Co-Authors: Sara Mina, Agnès Marot, Bernard Cimon, Maxime Fleury, Gérald Larcher, Jean-philippe Bouchara, Raymond RobertAbstract:Scedosporium boydii is an opportunistic filamentous fungus which may be responsible for a wide variety of infections in immunocompetent as well as immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF) and that may lead to allergic broncho-pulmonary mycoses, sensitization or respiratory infections. Upon microbial infection, host phagocytic cells release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by detoxification of the hydrogen peroxide. Here, we investigated the catalase equipment of S. boydii, one of the major pathogenic species in the S. apiospermum species complex. Three catalases were identified and the mycelial catalase A1 was purified to homogeneity by a three-step chromatographic process. This enzyme is a monofunctional tetrameric protein, of 460 kDa, consisting of four 82-kDa glycosylated subunits. The potential interest of this enzyme in serodiagnosis of S. apiospermum infections was then investigated by ELISA, using 64 sera from CF patients. Whatever the species involved in the S. apiospermum complex, sera from infected patients were clearly differentiated from sera from patients with an Aspergillus fumigatus infection, or from CF patients without clinical and biological signs of a fungal infection and without any fungus recovered from sputum samples. These results suggest that catalase A1 is a good candidate for the development of an immunoassay for serodiagnosis of infetions caused by the S. apiospermum complex in patients with CF.
Ghamrawi Sarah - One of the best experts on this subject based on the ideXlab platform.
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A multifaceted study of the cell wall changes during maturation and germination of the conidia in Scedosporium boydii
2014Co-Authors: Ghamrawi SarahAbstract:Les espèces du complexe Scedosporium apiospermum sont des agents pathogènes émergents qui se situent au deuxième rang parmi les champignons filamenteux rencontrés au cours de la mucoviscidose. Ils sont omniprésents et particulièrement rencontrés dans les zones polluées. En dépit de leur importance clinique, nos connaissances sur leur biologie moléculaire et leur physiologie restent limitées. Chez les champignons, la paroi constitue un bouclier protecteur face à des conditions environnementales défavorables, et joue un rôle essentiel dans la pathogénicité. Ici, nous avons étudié les changements dynamiques de la paroi des conidies de S. boydii, l’une des deux espèces majeures de ce complexe avec S. apiospermum, avec pour objectif d'identifier des facteurs de virulence potentiels. En utilisant une large variété de techniques, allant de la microscopie électronique à balayage ou à transmission à l’analyse protéomique des protéines à ancre glycosylphosphatidylinositol (GPI) en passant par la microélectrophorèse et la partition de phase, la cytométrie en flux, la microscopie de force atomique, la résonance paramagnétique électronique, ou encore des techniques moléculaires, nous avons mis en évidence diverses modifications qui se produisent dans la paroi pendant la maturation et la germination des conidies de S. boydii et nous avons identifié la DHN-mélanine ainsi qu'un nombre important de protéines à ancre GPI. Enfin, nous avons fourni la première séquence complète du génome de S. apiospermum qui appuierait les différents domaines de la recherche sur ces champignons que ce soit pour l’étude des mécanismes pathogènes ou pour des applications biotechnologiques.Species of the Scedosporium apiospermum complex are emerging human pathogens which rank the second, after Aspergillus fumigatus, among the filamentous fungi colonizing the airways of patients with cystic fibrosis. These fungi are ubiquitous in nature and particularly encountered in polluted areas. Despite their clinical relevance, our knowledge about their molecular biology and physiology remains rather limited. In fungi, the cell wall forms a protective shield against adverse environmental conditions, and therefore plays a key role in pathogenesis, which makes it an interesting target for antifungal drug development. Here, in an attempt to identify potential virulence factors, we investigated the dynamic changes of the cell wall of conidia in S. boydii, one of the main pathogenic species within this species complex with Scedosporium apiospermum. Using various techniques, ranging from scanning and transmission electron microscopy to proteomic analysis of glycosylphosphatidylinositol (GPI)- anchored proteins, through two-phase partitioning and microelectrophoresis, atomic force microscopy and chemical force spectroscopy, flow 5 cytometry, electron paramagnetic resonance and molecular techniques, we highlighted various modifications occurring in the cell wall during maturation and germination of S. boydii and we identified DHN-melanin as well as a substantial number of GPI-anchored proteins in the cell wall. Finally, we provided the first publicly available genome sequence of S. apiospermum that would support various research fields on these fungi whether for understanding their pathogenic mechanisms or for various biotechnological applications
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Modifications de la paroi au cours de la maturation et de la germination des conidies de Scedosporium boydii
HAL CCSD, 2014Co-Authors: Ghamrawi SarahAbstract:Species of the Scedosporium apiospermum complex are emerging human pathogens which rank the second, after Aspergillus fumigatus, among the filamentous fungi colonizing the airways of patients with cystic fibrosis. These fungi are ubiquitous in nature and particularly encountered in polluted areas. Despite their clinical relevance, our knowledge about their molecular biology and physiology remains rather limited. In fungi, the cell wall forms a protective shield against adverse environmental conditions, and therefore plays a key role in pathogenesis, which makes it an interesting target for antifungal drug development. Here, in an attempt to identify potential virulence factors, we investigated the dynamic changes of the cell wall of conidia in S. boydii, one of the main pathogenic species within this species complex with Scedosporium apiospermum. Using various techniques, ranging from scanning and transmission electron microscopy to proteomic analysis of glycosylphosphatidylinositol (GPI)- anchored proteins, through two-phase partitioning and microelectrophoresis, atomic force microscopy and chemical force spectroscopy, flow 5 cytometry, electron paramagnetic resonance and molecular techniques, we highlighted various modifications occurring in the cell wall during maturation and germination of S. boydii and we identified DHN-melanin as well as a substantial number of GPI-anchored proteins in the cell wall. Finally, we provided the first publicly available genome sequence of S. apiospermum that would support various research fields on these fungi whether for understanding their pathogenic mechanisms or for various biotechnological applications.Les espèces du complexe Scedosporium apiospermum sont des agents pathogènes émergents qui se situent au deuxième rang parmi les champignons filamenteux rencontrés au cours de la mucoviscidose. Ils sont omniprésents et particulièrement rencontrés dans les zones polluées. En dépit de leur importance clinique, nos connaissances sur leur biologie moléculaire et leur physiologie restent limitées. Chez les champignons, la paroi constitue un bouclier protecteur face à des conditions environnementales défavorables, et joue un rôle essentiel dans la pathogénicité. Ici, nous avons étudié les changements dynamiques de la paroi des conidies de S. boydii, l’une des deux espèces majeures de ce complexe avec S. apiospermum, avec pour objectif d'identifier des facteurs de virulence potentiels. En utilisant une large variété de techniques, allant de la microscopie électronique à balayage ou à transmission à l’analyse protéomique des protéines à ancre glycosylphosphatidylinositol (GPI) en passant par la microélectrophorèse et la partition de phase, la cytométrie en flux, la microscopie de force atomique, la résonance paramagnétique électronique, ou encore des techniques moléculaires, nous avons mis en évidence diverses modifications qui se produisent dans la paroi pendant la maturation et la germination des conidies de S. boydii et nous avons identifié la DHN-mélanine ainsi qu'un nombre important de protéines à ancre GPI. Enfin, nous avons fourni la première séquence complète du génome de S. apiospermum qui appuierait les différents domaines de la recherche sur ces champignons que ce soit pour l’étude des mécanismes pathogènes ou pour des applications biotechnologiques
Jean-philippe Bouchara - One of the best experts on this subject based on the ideXlab platform.
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Scedosporium boydii cata1 and sodc recombinant proteins new tools for serodiagnosis of Scedosporium infection of patients with cystic fibrosis
Diagnostic Microbiology and Infectious Disease, 2017Co-Authors: Sara Mina, Agnès Marot, Jean-philippe Bouchara, Cindy Staerck, Alphonse Calenda, Charlotte Godon, Maxime FleuryAbstract:Scedosporium species rank the second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF), after Aspergillus fumigatus. In CF, these fungi may cause various respiratory infections similar to those caused by A. fumigatus, including bronchitis and allergic broncho-pulmonary mycoses. Diagnosis of these infections relies on the detection of serum antibodies using crude antigenic extracts. However, many components of these extracts are common to Scedosporium and Aspergillus species, leading to cross-reactions. Here, 5 recombinant proteins from S. apiospermum or S. boydii were produced, and their value in serodiagnosis of Scedosporium infections was investigated by enzyme-linked immunosorbent assay. Two of them, corresponding to the Scedosporium catalase A1 or cytosolic Cu,Zn-superoxyde dismutase, allowed the detection of Scedosporium infection, and the differentiation with an Aspergillus infection. These recombinant proteins therefore may serve as a basis for the development of a standardized serological test.
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the secreted polyketide boydone a is responsible for the anti staphylococcus aureus activity of Scedosporium boydii
Fems Microbiology Letters, 2017Co-Authors: Cindy Staerck, Gérald Larcher, Jean-philippe Bouchara, Anne Landreau, Gaetan Herbette, Catherine Roullier, Samuel Bertrand, Benjamin Siegler, Maxime FleuryAbstract:Usually living as a soil saprophyte, the filamentous fungus Scedosporium boydii may also cause various infections in human. Particularly, it is one of the major causative agents of fungal colonization of the airways in patients with cystic fibrosis (CF). To compete with other microorganisms in the environment, fungi have evolved sophisticated strategies, including the production of secondary metabolites with antimicrobial activity that may also help them to establish successfully within the respiratory tract of receptive hosts. Here, the culture filtrate from a human pathogenic strain of S. boydii was investigated searching for an antibacterial activity, mainly against the major CF bacterial pathogens. A high antibacterial activity against Staphylococcus aureus, including methicillin-resistant strains of this species, was observed. Bio-guided fractionation and analysis of the active fractions by nuclear magnetic resonance or by high-performance liquid chromatography and high-resolution electrospray ionization-mass spectrometry allowed us to identify boydone A as responsible for this antibacterial activity. Together, these results suggest that this six-membered cyclic polyketide could be one of the virulence factors of the fungus. Genes involved in the synthesis of this secreted metabolite are currently being identified in order to confirm the role of this polyketide in pathogenesis.
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draft genome sequence of the human pathogenic fungus Scedosporium boydii
Genome Announcements, 2017Co-Authors: Ludovic Duvaux, Jean-philippe Bouchara, Jason Shiller, Patrick Vandeputte, Thomas Duge De Bernonville, Christopher R Thornton, Nicolas Papon, Bruno Le Cam, Amandine GasteboisAbstract:The opportunistic fungal pathogen Scedosporium boydii is the most common Scedosporium species in French patients with cystic fibrosis. Here we present the first genome report for S. boydii, providing a resource which may enable the elucidation of the pathogenic mechanisms in this species.
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Purification and characterization of a mycelial catalase from Scedosporium boydii, a useful tool for specific antibody detection in patients with cystic fibrosis
Clinical and Vaccine Immunology, 2015Co-Authors: Sara Mina, Agnès Marot, Bernard Cimon, Maxime Fleury, Gérald Larcher, Jean-philippe Bouchara, Raymond RobertAbstract:Scedosporium boydii is an opportunistic filamentous fungus which may be responsible for a wide variety of infections in immunocompetent as well as immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF) and that may lead to allergic broncho-pulmonary mycoses, sensitization or respiratory infections. Upon microbial infection, host phagocytic cells release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by detoxification of the hydrogen peroxide. Here, we investigated the catalase equipment of S. boydii, one of the major pathogenic species in the S. apiospermum species complex. Three catalases were identified and the mycelial catalase A1 was purified to homogeneity by a three-step chromatographic process. This enzyme is a monofunctional tetrameric protein, of 460 kDa, consisting of four 82-kDa glycosylated subunits. The potential interest of this enzyme in serodiagnosis of S. apiospermum infections was then investigated by ELISA, using 64 sera from CF patients. Whatever the species involved in the S. apiospermum complex, sera from infected patients were clearly differentiated from sera from patients with an Aspergillus fumigatus infection, or from CF patients without clinical and biological signs of a fungal infection and without any fungus recovered from sputum samples. These results suggest that catalase A1 is a good candidate for the development of an immunoassay for serodiagnosis of infetions caused by the S. apiospermum complex in patients with CF.
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A Multifaceted Study of Scedosporium boydii Cell Wall Changes during Germination and Identification of GPI-Anchored Proteins
2015Co-Authors: Sarah Ghamrawi, Agnès Marot, Patrick Vandeputte, Amandine Gastebois, Agata Zykwinska, Guillaume Mabilleau, Stéphane Cuenot, Jean-philippe BoucharaAbstract:Scedosporium boydii is a pathogenic filamentous fungus that causes a wide range of human infections, notably respiratory infections in patients with cystic fibrosis. The development of new therapeutic strategies targeting S. boydii necessitates a better understanding of the physiology of this fungus and the identification of new molecular targets. In this work, we studied the conidium-to-germ tube transition using a variety of techniques including scanning and transmission electron microscopy, atomic force microscopy, two-phase partitioning, microelectrophoresis and cationized ferritin labeling, chemical force spectroscopy, lectin labeling, and nanoLC-MS/MS for cell wall GPI-anchored protein analysis. We demonstrated that the cell wall undergoes structural changes with germination accompanied with a lower hydrophobicity, electrostatic charge and binding capacity to cationized ferritin. Changes during germination also included a higher accessibility of some cell wall polysaccharides to lectins and less CH3/CH3 interactions (hydrophobic adhesion forces mainly due to glycoproteins). We also extracted and identified 20 GPI-anchored proteins from the cell wall of S. boydii, among which one was detected only in the conidial wall extract and 12 only in the mycelial wall extract. The identified sequences belonged to protein families involved in virulence in other fungi like Gelp/Gasp, Crhp, Bglp/Bgtp families and a superoxide dismutase. These results highlighted the cell wall remodeling during germination in S. boydii with the identification of a substantial number of cell wall GPI-anchored conidial or hyphal specific proteins, which provides a basis to investigate the role of these molecules in the host-pathogen interaction and fungal virulence.
