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In vitro characteristics of an airway barrier-disrupting factor secreted by Staphylococcus aureus.
Murphy, Jae; Ramezanpour, Mahnaz; Drilling, Amanda; Roscioli, Eugene; Psaltis, Alkis James; Wormald, Peter-John; Vreugde, Sarah.
Afiliação
  • Murphy J; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
  • Ramezanpour M; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
  • Drilling A; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
  • Roscioli E; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
  • Psaltis AJ; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
  • Wormald PJ; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
  • Vreugde S; Department of Otolaryngology-Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
Int Forum Allergy Rhinol ; 9(2): 187-196, 2019 02.
Article em En | MEDLINE | ID: mdl-30431711
BACKGROUND: Staphylococcus aureus is a major contributor to the pathophysiology of chronic rhinosinusitis (CRS). Previous research has shown that S. aureus-secreted products disrupt the airway barrier. METHODS: S. aureus ATCC 13565 and 25923 strains were grown at exponential, postexponential, and stationary phases. Microbial conditioned media (CM) was collected from the cultures and ultrafiltered (UF). Liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was performed on the UF-CM. UF-CM was subjected to heat and protease treatment, size fractionation, and ultracentrifugation (UC) separation. Human nasal epithelial cells grown at air-liquid interface (HNEC-ALI) cultures were exposed to purified alpha hemolysin (Hla), staphylococcal enterotoxin A (SEA), lipoteichoic acid (LTA), and UF-CM. Barrier function outcomes were measured by transepithelial electrical resistance (TEER) and apparent permeability (Papp). UC fraction exposed cultures were subjected to immunofluorescence microscopy for tight junction (TJ) protein zonula occludens-1 (ZO-1). RESULTS: LC-ESI-MS/MS identified 107 proteins, with Hla being most abundant. Hla, SEA, and LTA did not alter the HNEC-ALI barrier as measured by TEER or Papp. Barrier disruption caused by UF-CM peaked in the postexponential phase, was sensitive to heat and protease treatment, >30-kDa in size, and enriched in the UC fraction. HNEC-ALI exposed to UF-CM and UC demonstrated loss of ZO-1 localization. CONCLUSION: These results suggest that the S. aureus factor responsible for TJ disruption in HNEC-ALI cultures is either a protein-macromolecule or a combination of secreted factors. The product is enriched in the UC fraction, suggesting it is associated with large structures such as membrane components or vesicles.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Sinusite / Infecções Estafilocócicas / Staphylococcus aureus / Rinite / Meios de Cultivo Condicionados / Junções Íntimas / Mucosa Nasal Tipo de estudo: Prognostic_studies Limite: Humans Idioma: En Ano de publicação: 2019 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Sinusite / Infecções Estafilocócicas / Staphylococcus aureus / Rinite / Meios de Cultivo Condicionados / Junções Íntimas / Mucosa Nasal Tipo de estudo: Prognostic_studies Limite: Humans Idioma: En Ano de publicação: 2019 Tipo de documento: Article