Characterization of the adherence of Clostridium difficile spores: The integrity of the outermost layer affects adherence properties of spores of the epidemic strain R20291 to components of the intestinal mucosa

Paola Mora-Uribe, Camila Miranda-Cárdenas, Pablo Castro-Córdova, Fernando Gil, Iván Calderón, Juan A. Fuentes, Paula I. Rodas, Saeed Banawas, Mahfuzur R. Sarker, Daniel Paredes-Sabja

Resultado de la investigación: Contribución a la publicaciónArticle

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Resumen

Clostridium difficile is the causative agent of the most frequently reported nosocomial diarrhea worldwide. The high incidence of recurrent infection is the main clinical challenge of C. difficile infections (CDI). Formation of C. difficile spores of the epidemic strain R20291 has been shown to be essential for recurrent infection and transmission of the disease in a mouse model. However, the underlying mechanisms of how these spores persist in the colonic environment remains unclear. In this work, we characterized the adherence properties of epidemic R20291 spores to components of the intestinal mucosa, and we assessed the role of the exosporium integrity in the adherence properties by using cdeC mutant spores with a defective exosporium layer. Our results showed that spores and vegetative cells of the epidemic R20291 strain adhered at high levels to monolayers of Caco-2 cells and mucin. Transmission electron micrographs of Caco-2 cells demonstrated that the hair-like projections on the surface of R20291 spores are in close proximity with the plasma membrane and microvilli of undifferentiated and differentiated monolayers of Caco-2 cells. Competitive-binding assay in differentiated Caco-2 cells suggests that spore-adherence is mediated by specific binding sites. By using spores of a cdeC mutant we demonstrated that the integrity of the exosporium layer determines the affinity of adherence of C. difficile spores to Caco-2 cells and mucin. Binding of fibronectin and vitronectin to the spore surface was concentration-dependent, and depending on the concentration, spore-adherence to Caco-2 cells was enhanced. In the presence of an aberrantly-assembled exosporium (cdeC spores), binding of fibronectin, but not vitronectin, was increased. Notably, independent of the exosporium integrity, only a fraction of the spores had fibronectin and vitronectin molecules binding to their surface. Collectively, these results demonstrate that the integrity of the exosporium layer of strain R20291 contributes to selective spore adherence to components of the intestinal mucosa.

Idioma originalEnglish
Número de artículo099
PublicaciónFrontiers in cellular and infection microbiology
Volumen6
Número de ediciónSEP
Identificadores de objetos digitales
EstadoPublished - 22 sep 2016

Huella dactilar

Clostridium difficile
Intestinal Mucosa
Spores
Caco-2 Cells
Vitronectin
Fibronectins
Mucins
Clostridium Infections
Infectious Disease Transmission
Competitive Binding
Microvilli
Hair
Diarrhea
Binding Sites
Cell Membrane
Electrons
Incidence
Infection

Keywords

    ASJC Scopus subject areas

    • Microbiology
    • Immunology
    • Medicine(all)
    • Microbiology (medical)
    • Infectious Diseases

    Citar esto

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    title = "Characterization of the adherence of Clostridium difficile spores: The integrity of the outermost layer affects adherence properties of spores of the epidemic strain R20291 to components of the intestinal mucosa",
    abstract = "Clostridium difficile is the causative agent of the most frequently reported nosocomial diarrhea worldwide. The high incidence of recurrent infection is the main clinical challenge of C. difficile infections (CDI). Formation of C. difficile spores of the epidemic strain R20291 has been shown to be essential for recurrent infection and transmission of the disease in a mouse model. However, the underlying mechanisms of how these spores persist in the colonic environment remains unclear. In this work, we characterized the adherence properties of epidemic R20291 spores to components of the intestinal mucosa, and we assessed the role of the exosporium integrity in the adherence properties by using cdeC mutant spores with a defective exosporium layer. Our results showed that spores and vegetative cells of the epidemic R20291 strain adhered at high levels to monolayers of Caco-2 cells and mucin. Transmission electron micrographs of Caco-2 cells demonstrated that the hair-like projections on the surface of R20291 spores are in close proximity with the plasma membrane and microvilli of undifferentiated and differentiated monolayers of Caco-2 cells. Competitive-binding assay in differentiated Caco-2 cells suggests that spore-adherence is mediated by specific binding sites. By using spores of a cdeC mutant we demonstrated that the integrity of the exosporium layer determines the affinity of adherence of C. difficile spores to Caco-2 cells and mucin. Binding of fibronectin and vitronectin to the spore surface was concentration-dependent, and depending on the concentration, spore-adherence to Caco-2 cells was enhanced. In the presence of an aberrantly-assembled exosporium (cdeC spores), binding of fibronectin, but not vitronectin, was increased. Notably, independent of the exosporium integrity, only a fraction of the spores had fibronectin and vitronectin molecules binding to their surface. Collectively, these results demonstrate that the integrity of the exosporium layer of strain R20291 contributes to selective spore adherence to components of the intestinal mucosa.",
    keywords = "BclA, C. difficile spores, CdeC, Exosporium, Spore adherence",
    author = "Paola Mora-Uribe and Camila Miranda-Cárdenas and Pablo Castro-Córdova and Fernando Gil and Iván Calderón and Fuentes, {Juan A.} and Rodas, {Paula I.} and Saeed Banawas and Sarker, {Mahfuzur R.} and Daniel Paredes-Sabja",
    year = "2016",
    month = "9",
    doi = "10.3389/fcimb.2016.00099",
    volume = "6",
    journal = "Frontiers in cellular and infection microbiology",
    issn = "2235-2988",
    publisher = "Frontiers Media S.A.",
    number = "SEP",

    }

    TY - JOUR

    T1 - Characterization of the adherence of Clostridium difficile spores

    T2 - Frontiers in cellular and infection microbiology

    AU - Mora-Uribe,Paola

    AU - Miranda-Cárdenas,Camila

    AU - Castro-Córdova,Pablo

    AU - Gil,Fernando

    AU - Calderón,Iván

    AU - Fuentes,Juan A.

    AU - Rodas,Paula I.

    AU - Banawas,Saeed

    AU - Sarker,Mahfuzur R.

    AU - Paredes-Sabja,Daniel

    PY - 2016/9/22

    Y1 - 2016/9/22

    N2 - Clostridium difficile is the causative agent of the most frequently reported nosocomial diarrhea worldwide. The high incidence of recurrent infection is the main clinical challenge of C. difficile infections (CDI). Formation of C. difficile spores of the epidemic strain R20291 has been shown to be essential for recurrent infection and transmission of the disease in a mouse model. However, the underlying mechanisms of how these spores persist in the colonic environment remains unclear. In this work, we characterized the adherence properties of epidemic R20291 spores to components of the intestinal mucosa, and we assessed the role of the exosporium integrity in the adherence properties by using cdeC mutant spores with a defective exosporium layer. Our results showed that spores and vegetative cells of the epidemic R20291 strain adhered at high levels to monolayers of Caco-2 cells and mucin. Transmission electron micrographs of Caco-2 cells demonstrated that the hair-like projections on the surface of R20291 spores are in close proximity with the plasma membrane and microvilli of undifferentiated and differentiated monolayers of Caco-2 cells. Competitive-binding assay in differentiated Caco-2 cells suggests that spore-adherence is mediated by specific binding sites. By using spores of a cdeC mutant we demonstrated that the integrity of the exosporium layer determines the affinity of adherence of C. difficile spores to Caco-2 cells and mucin. Binding of fibronectin and vitronectin to the spore surface was concentration-dependent, and depending on the concentration, spore-adherence to Caco-2 cells was enhanced. In the presence of an aberrantly-assembled exosporium (cdeC spores), binding of fibronectin, but not vitronectin, was increased. Notably, independent of the exosporium integrity, only a fraction of the spores had fibronectin and vitronectin molecules binding to their surface. Collectively, these results demonstrate that the integrity of the exosporium layer of strain R20291 contributes to selective spore adherence to components of the intestinal mucosa.

    AB - Clostridium difficile is the causative agent of the most frequently reported nosocomial diarrhea worldwide. The high incidence of recurrent infection is the main clinical challenge of C. difficile infections (CDI). Formation of C. difficile spores of the epidemic strain R20291 has been shown to be essential for recurrent infection and transmission of the disease in a mouse model. However, the underlying mechanisms of how these spores persist in the colonic environment remains unclear. In this work, we characterized the adherence properties of epidemic R20291 spores to components of the intestinal mucosa, and we assessed the role of the exosporium integrity in the adherence properties by using cdeC mutant spores with a defective exosporium layer. Our results showed that spores and vegetative cells of the epidemic R20291 strain adhered at high levels to monolayers of Caco-2 cells and mucin. Transmission electron micrographs of Caco-2 cells demonstrated that the hair-like projections on the surface of R20291 spores are in close proximity with the plasma membrane and microvilli of undifferentiated and differentiated monolayers of Caco-2 cells. Competitive-binding assay in differentiated Caco-2 cells suggests that spore-adherence is mediated by specific binding sites. By using spores of a cdeC mutant we demonstrated that the integrity of the exosporium layer determines the affinity of adherence of C. difficile spores to Caco-2 cells and mucin. Binding of fibronectin and vitronectin to the spore surface was concentration-dependent, and depending on the concentration, spore-adherence to Caco-2 cells was enhanced. In the presence of an aberrantly-assembled exosporium (cdeC spores), binding of fibronectin, but not vitronectin, was increased. Notably, independent of the exosporium integrity, only a fraction of the spores had fibronectin and vitronectin molecules binding to their surface. Collectively, these results demonstrate that the integrity of the exosporium layer of strain R20291 contributes to selective spore adherence to components of the intestinal mucosa.

    KW - BclA

    KW - C. difficile spores

    KW - CdeC

    KW - Exosporium

    KW - Spore adherence

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    U2 - 10.3389/fcimb.2016.00099

    DO - 10.3389/fcimb.2016.00099

    M3 - Article

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    JO - Frontiers in cellular and infection microbiology

    JF - Frontiers in cellular and infection microbiology

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