Staphylococcus aureus impairs the airway epithelial barrier in vitro

Background Chronic rhinosinusitis (CRS) is a cluster of disorders that result in sinonasal mucosal inflammation. Staphylococcus aureus (S. aureus) is associated with severe and recalcitrant CRS. The purpose of our study was to investigate the effect of S. aureus on respiratory epithelial barrier str...

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Veröffentlicht in:International forum of allergy & rhinology 2015-06, Vol.5 (6), p.551-556
Hauptverfasser: Malik, Zacki, Roscioli, Eugene, Murphy, Jae, Ou, Judy, Bassiouni, Ahmed, Wormald, Peter-John, Vreugde, Sarah
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container_issue 6
container_start_page 551
container_title International forum of allergy & rhinology
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creator Malik, Zacki
Roscioli, Eugene
Murphy, Jae
Ou, Judy
Bassiouni, Ahmed
Wormald, Peter-John
Vreugde, Sarah
description Background Chronic rhinosinusitis (CRS) is a cluster of disorders that result in sinonasal mucosal inflammation. Staphylococcus aureus (S. aureus) is associated with severe and recalcitrant CRS. The purpose of our study was to investigate the effect of S. aureus on respiratory epithelial barrier structure and function. Methods Conditioned media from S. aureus reference strains (American Type Culture Collection [ATCC] 13565, 14458, and 25923) was applied to air‐liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs) and transepithelial electrical resistance (TEER) was measured to assess cell‐to‐cell integrity. Electron microscopy was used to gauge the ciliated area and tight junctions (TJs). Additionally, the expression of the TJ protein zona occludens‐1 (ZO‐1) was examined via immunofluorescence. Statistical analysis was performed using analysis of variance (ANOVA) with pairwise Bonferroni‐adjusted t tests. Results Secreted products applied to ALI cultures from S. aureus strain 13565 caused a concentration‐dependent decline in electrical impedance compared to controls and reference strains 14458 and 25923 (p < 0.001). Electron microscopy showed a distinct separation between adjacent cells apically, in the region of TJs. The ciliated area was not affected; however, ZO‐1 expression became discontinuous in HNECs exposed to the 13565 strain's conditioned media. Conclusion Conditioned media of the S. aureus strain 13565 damages the airway epithelium by disrupting the TJs between primary HNECs grown at an ALI. These findings suggest that strain‐specific S. aureus–secreted product(s) compromise epithelial barrier function, which may constitute 1 of the roles played by S. aureus in the pathophysiology of recalcitrant CRS. Further research is required to uncover the relevant molecular mechanisms.
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Staphylococcus aureus (S. aureus) is associated with severe and recalcitrant CRS. The purpose of our study was to investigate the effect of S. aureus on respiratory epithelial barrier structure and function. Methods Conditioned media from S. aureus reference strains (American Type Culture Collection [ATCC] 13565, 14458, and 25923) was applied to air‐liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs) and transepithelial electrical resistance (TEER) was measured to assess cell‐to‐cell integrity. Electron microscopy was used to gauge the ciliated area and tight junctions (TJs). Additionally, the expression of the TJ protein zona occludens‐1 (ZO‐1) was examined via immunofluorescence. Statistical analysis was performed using analysis of variance (ANOVA) with pairwise Bonferroni‐adjusted t tests. Results Secreted products applied to ALI cultures from S. aureus strain 13565 caused a concentration‐dependent decline in electrical impedance compared to controls and reference strains 14458 and 25923 (p &lt; 0.001). Electron microscopy showed a distinct separation between adjacent cells apically, in the region of TJs. The ciliated area was not affected; however, ZO‐1 expression became discontinuous in HNECs exposed to the 13565 strain's conditioned media. Conclusion Conditioned media of the S. aureus strain 13565 damages the airway epithelium by disrupting the TJs between primary HNECs grown at an ALI. These findings suggest that strain‐specific S. aureus–secreted product(s) compromise epithelial barrier function, which may constitute 1 of the roles played by S. aureus in the pathophysiology of recalcitrant CRS. Further research is required to uncover the relevant molecular mechanisms.</description><identifier>ISSN: 2042-6976</identifier><identifier>EISSN: 2042-6984</identifier><identifier>DOI: 10.1002/alr.21517</identifier><identifier>PMID: 25821008</identifier><language>eng</language><publisher>United States: Blackwell Publishing Ltd</publisher><subject>air-liquid interface ; Bacteriology ; Cells, Cultured ; chronic rhinosinusitis ; Culture Media, Conditioned ; Electric Impedance ; epithelium ; Fluorescent Antibody Technique, Indirect ; Humans ; Intercellular Junctions - metabolism ; Microscopy ; Microscopy, Electron ; Nasal Cavity - cytology ; Nasal Mucosa - metabolism ; Nasal Mucosa - microbiology ; Nasal Mucosa - pathology ; Staphylococcus aureus - physiology ; Tight Junctions ; Variance analysis ; Zonula Occludens-1 Protein - metabolism</subject><ispartof>International forum of allergy &amp; rhinology, 2015-06, Vol.5 (6), p.551-556</ispartof><rights>2015 ARS‐AAOA, LLC</rights><rights>2015 ARS-AAOA, LLC.</rights><rights>2015 ARS-AAOA, LLC</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4577-f9966e3cba1d5955121b4ca08f04560764d73a46aab64af40d8058c02c12fb1b3</citedby><cites>FETCH-LOGICAL-c4577-f9966e3cba1d5955121b4ca08f04560764d73a46aab64af40d8058c02c12fb1b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Falr.21517$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Falr.21517$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,777,781,1413,27906,27907,45556,45557</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25821008$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Malik, Zacki</creatorcontrib><creatorcontrib>Roscioli, Eugene</creatorcontrib><creatorcontrib>Murphy, Jae</creatorcontrib><creatorcontrib>Ou, Judy</creatorcontrib><creatorcontrib>Bassiouni, Ahmed</creatorcontrib><creatorcontrib>Wormald, Peter-John</creatorcontrib><creatorcontrib>Vreugde, Sarah</creatorcontrib><title>Staphylococcus aureus impairs the airway epithelial barrier in vitro</title><title>International forum of allergy &amp; rhinology</title><addtitle>International Forum of Allergy and Rhinology</addtitle><description>Background Chronic rhinosinusitis (CRS) is a cluster of disorders that result in sinonasal mucosal inflammation. Staphylococcus aureus (S. aureus) is associated with severe and recalcitrant CRS. The purpose of our study was to investigate the effect of S. aureus on respiratory epithelial barrier structure and function. Methods Conditioned media from S. aureus reference strains (American Type Culture Collection [ATCC] 13565, 14458, and 25923) was applied to air‐liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs) and transepithelial electrical resistance (TEER) was measured to assess cell‐to‐cell integrity. Electron microscopy was used to gauge the ciliated area and tight junctions (TJs). Additionally, the expression of the TJ protein zona occludens‐1 (ZO‐1) was examined via immunofluorescence. Statistical analysis was performed using analysis of variance (ANOVA) with pairwise Bonferroni‐adjusted t tests. Results Secreted products applied to ALI cultures from S. aureus strain 13565 caused a concentration‐dependent decline in electrical impedance compared to controls and reference strains 14458 and 25923 (p &lt; 0.001). Electron microscopy showed a distinct separation between adjacent cells apically, in the region of TJs. The ciliated area was not affected; however, ZO‐1 expression became discontinuous in HNECs exposed to the 13565 strain's conditioned media. Conclusion Conditioned media of the S. aureus strain 13565 damages the airway epithelium by disrupting the TJs between primary HNECs grown at an ALI. These findings suggest that strain‐specific S. aureus–secreted product(s) compromise epithelial barrier function, which may constitute 1 of the roles played by S. aureus in the pathophysiology of recalcitrant CRS. Further research is required to uncover the relevant molecular mechanisms.</description><subject>air-liquid interface</subject><subject>Bacteriology</subject><subject>Cells, Cultured</subject><subject>chronic rhinosinusitis</subject><subject>Culture Media, Conditioned</subject><subject>Electric Impedance</subject><subject>epithelium</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Humans</subject><subject>Intercellular Junctions - metabolism</subject><subject>Microscopy</subject><subject>Microscopy, Electron</subject><subject>Nasal Cavity - cytology</subject><subject>Nasal Mucosa - metabolism</subject><subject>Nasal Mucosa - microbiology</subject><subject>Nasal Mucosa - pathology</subject><subject>Staphylococcus aureus - physiology</subject><subject>Tight Junctions</subject><subject>Variance analysis</subject><subject>Zonula Occludens-1 Protein - metabolism</subject><issn>2042-6976</issn><issn>2042-6984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kMtOwzAQRS0EAlRY8AMoEhtYpLUdP9IlolAQbUE8BDtr4jrCkDTBTij9e1wKXSAxm5mRzr0zuggdENwlGNMeFK5LCSdyA-1SzGgs-inbXM9S7KB9719xKE544LbRDuUpDeJ0Fw3uG6hfFkWlK61bH0HrTGi2rME6HzUvJgrDHBaRqW3YCgtFlIFz1rjIzqIP27hqD23lUHiz_9M76PHi_OHsMh7dDK_OTkexZlzKOO_3hTCJzoBMeT-8QknGNOA0x4wLLAWbygSYAMgEg5zhaYp5qjHVhOYZyZIOOl751q56b41vVGm9NkUBM1O1XhGRyoRJSUlAj_6gr1XrZuG7JRWOLaFAnawo7SrvnclV7WwJbqEIVst0VUhXfacb2MMfxzYrzXRN_mYZgN4KmNvCLP53Uqeju1_LeKWwvjGfawW4NyVkIrl6mgzV8_VkQMe3Y5UkX7eMkTE</recordid><startdate>201506</startdate><enddate>201506</enddate><creator>Malik, Zacki</creator><creator>Roscioli, Eugene</creator><creator>Murphy, Jae</creator><creator>Ou, Judy</creator><creator>Bassiouni, Ahmed</creator><creator>Wormald, Peter-John</creator><creator>Vreugde, Sarah</creator><general>Blackwell Publishing Ltd</general><general>Wiley Subscription Services, Inc</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope><scope>7X8</scope></search><sort><creationdate>201506</creationdate><title>Staphylococcus aureus impairs the airway epithelial barrier in vitro</title><author>Malik, Zacki ; Roscioli, Eugene ; Murphy, Jae ; Ou, Judy ; Bassiouni, Ahmed ; Wormald, Peter-John ; Vreugde, Sarah</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4577-f9966e3cba1d5955121b4ca08f04560764d73a46aab64af40d8058c02c12fb1b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>air-liquid interface</topic><topic>Bacteriology</topic><topic>Cells, Cultured</topic><topic>chronic rhinosinusitis</topic><topic>Culture Media, Conditioned</topic><topic>Electric Impedance</topic><topic>epithelium</topic><topic>Fluorescent Antibody Technique, Indirect</topic><topic>Humans</topic><topic>Intercellular Junctions - metabolism</topic><topic>Microscopy</topic><topic>Microscopy, Electron</topic><topic>Nasal Cavity - cytology</topic><topic>Nasal Mucosa - metabolism</topic><topic>Nasal Mucosa - microbiology</topic><topic>Nasal Mucosa - pathology</topic><topic>Staphylococcus aureus - physiology</topic><topic>Tight Junctions</topic><topic>Variance analysis</topic><topic>Zonula Occludens-1 Protein - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Malik, Zacki</creatorcontrib><creatorcontrib>Roscioli, Eugene</creatorcontrib><creatorcontrib>Murphy, Jae</creatorcontrib><creatorcontrib>Ou, Judy</creatorcontrib><creatorcontrib>Bassiouni, Ahmed</creatorcontrib><creatorcontrib>Wormald, Peter-John</creatorcontrib><creatorcontrib>Vreugde, Sarah</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>International forum of allergy &amp; rhinology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Malik, Zacki</au><au>Roscioli, Eugene</au><au>Murphy, Jae</au><au>Ou, Judy</au><au>Bassiouni, Ahmed</au><au>Wormald, Peter-John</au><au>Vreugde, Sarah</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Staphylococcus aureus impairs the airway epithelial barrier in vitro</atitle><jtitle>International forum of allergy &amp; rhinology</jtitle><addtitle>International Forum of Allergy and Rhinology</addtitle><date>2015-06</date><risdate>2015</risdate><volume>5</volume><issue>6</issue><spage>551</spage><epage>556</epage><pages>551-556</pages><issn>2042-6976</issn><eissn>2042-6984</eissn><abstract>Background Chronic rhinosinusitis (CRS) is a cluster of disorders that result in sinonasal mucosal inflammation. Staphylococcus aureus (S. aureus) is associated with severe and recalcitrant CRS. The purpose of our study was to investigate the effect of S. aureus on respiratory epithelial barrier structure and function. Methods Conditioned media from S. aureus reference strains (American Type Culture Collection [ATCC] 13565, 14458, and 25923) was applied to air‐liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs) and transepithelial electrical resistance (TEER) was measured to assess cell‐to‐cell integrity. Electron microscopy was used to gauge the ciliated area and tight junctions (TJs). Additionally, the expression of the TJ protein zona occludens‐1 (ZO‐1) was examined via immunofluorescence. Statistical analysis was performed using analysis of variance (ANOVA) with pairwise Bonferroni‐adjusted t tests. Results Secreted products applied to ALI cultures from S. aureus strain 13565 caused a concentration‐dependent decline in electrical impedance compared to controls and reference strains 14458 and 25923 (p &lt; 0.001). Electron microscopy showed a distinct separation between adjacent cells apically, in the region of TJs. The ciliated area was not affected; however, ZO‐1 expression became discontinuous in HNECs exposed to the 13565 strain's conditioned media. Conclusion Conditioned media of the S. aureus strain 13565 damages the airway epithelium by disrupting the TJs between primary HNECs grown at an ALI. These findings suggest that strain‐specific S. aureus–secreted product(s) compromise epithelial barrier function, which may constitute 1 of the roles played by S. aureus in the pathophysiology of recalcitrant CRS. Further research is required to uncover the relevant molecular mechanisms.</abstract><cop>United States</cop><pub>Blackwell Publishing Ltd</pub><pmid>25821008</pmid><doi>10.1002/alr.21517</doi><tpages>6</tpages></addata></record>
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subjects air-liquid interface
Bacteriology
Cells, Cultured
chronic rhinosinusitis
Culture Media, Conditioned
Electric Impedance
epithelium
Fluorescent Antibody Technique, Indirect
Humans
Intercellular Junctions - metabolism
Microscopy
Microscopy, Electron
Nasal Cavity - cytology
Nasal Mucosa - metabolism
Nasal Mucosa - microbiology
Nasal Mucosa - pathology
Staphylococcus aureus - physiology
Tight Junctions
Variance analysis
Zonula Occludens-1 Protein - metabolism
title Staphylococcus aureus impairs the airway epithelial barrier in vitro
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