EGTA treatment of human airways in vitro unmasks M1/MUC5AC mucin in submucosal glands
Mucin staining can be used to evaluate secretory activity of human airways. However, mucin epitopes may be masked by physicochemical properties of the secretions. The aim of this investigation was to examine the effects of the calcium chelator, ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N...
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| Veröffentlicht in: | The European respiratory journal 2001-07, Vol.18 (1), p.176-183 |
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| creator | Roger, P Gascard, J-P Bara, J Dulmet, E Brink, C |
| description | Mucin staining can be used to evaluate secretory activity of human airways. However, mucin epitopes may be masked by physicochemical properties of the secretions. The aim of this investigation was to examine the effects of the calcium chelator, ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) on the detection of M1/MUC5AC mucin in isolated human bronchial preparations. Immunohistochemical investigation and immunoradiometric assays with anti-M1 monoclonal antibodies (Mabs) were used to detect M1/MUC5AC mucin derived from bronchial preparations with an intact surface epithelium, or in tissues where the epithelium had been removed (rubbed preparations). The Mabs labelled both epithelial goblet cells and submucosal glandular cells in EGTA (4 mM)-exposed bronchial preparations, while only goblet cells were stained in EGTA (0.4 mM)-exposed tissues. The quantities of M1/MUC5AC mucin detected in either the bronchial fluids derived from EGTA (4 mM)-exposed intact and rubbed preparations or in bronchial fluids treated with EGTA (4 mM) were significantly increased by two-fold when compared with untreated control values (p |
| doi_str_mv | 10.1183/09031936.01.00105301 |
| format | Article |
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However, mucin epitopes may be masked by physicochemical properties of the secretions. The aim of this investigation was to examine the effects of the calcium chelator, ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) on the detection of M1/MUC5AC mucin in isolated human bronchial preparations. Immunohistochemical investigation and immunoradiometric assays with anti-M1 monoclonal antibodies (Mabs) were used to detect M1/MUC5AC mucin derived from bronchial preparations with an intact surface epithelium, or in tissues where the epithelium had been removed (rubbed preparations). The Mabs labelled both epithelial goblet cells and submucosal glandular cells in EGTA (4 mM)-exposed bronchial preparations, while only goblet cells were stained in EGTA (0.4 mM)-exposed tissues. The quantities of M1/MUC5AC mucin detected in either the bronchial fluids derived from EGTA (4 mM)-exposed intact and rubbed preparations or in bronchial fluids treated with EGTA (4 mM) were significantly increased by two-fold when compared with untreated control values (p<0.001). In addition, lactate dehydrogenase (LDH) activity and protein measurements were unaltered during exposure of human airways to EGTA (4 mM) suggesting that this treatment did not affect tissue viability. These results provide evidence that ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (4 mM) facilitates the detection of M1/MUC5AC mucin by altering the physicochemical properties of respiratory mucin, thereby exposing epitopes with which anti-M1 monoclonal antibodies are reactive. This will allow more accurate measurement of secretory activity in human airways in vitro.</description><identifier>ISSN: 0903-1936</identifier><identifier>EISSN: 1399-3003</identifier><identifier>DOI: 10.1183/09031936.01.00105301</identifier><identifier>PMID: 11510791</identifier><language>eng</language><publisher>Leeds: Eur Respiratory Soc</publisher><subject>Biological and medical sciences ; Bronchi - pathology ; Egtazic Acid - pharmacology ; Humans ; In Vitro Techniques ; Investigative techniques, diagnostic techniques (general aspects) ; Lung Neoplasms - pathology ; Medical sciences ; Mucin 5AC ; Mucins - analysis ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Respiratory Mucosa - pathology ; Respiratory system ; Sensitivity and Specificity</subject><ispartof>The European respiratory journal, 2001-07, Vol.18 (1), p.176-183</ispartof><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c363t-30fa13e4e840111b4dda3f5c552e932b0b02f95edf69eb1289f27cb0cff2b2e03</citedby><cites>FETCH-LOGICAL-c363t-30fa13e4e840111b4dda3f5c552e932b0b02f95edf69eb1289f27cb0cff2b2e03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,782,786,27931,27932</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14065319$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11510791$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Roger, P</creatorcontrib><creatorcontrib>Gascard, J-P</creatorcontrib><creatorcontrib>Bara, J</creatorcontrib><creatorcontrib>Dulmet, E</creatorcontrib><creatorcontrib>Brink, C</creatorcontrib><title>EGTA treatment of human airways in vitro unmasks M1/MUC5AC mucin in submucosal glands</title><title>The European respiratory journal</title><addtitle>Eur Respir J</addtitle><description>Mucin staining can be used to evaluate secretory activity of human airways. However, mucin epitopes may be masked by physicochemical properties of the secretions. The aim of this investigation was to examine the effects of the calcium chelator, ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) on the detection of M1/MUC5AC mucin in isolated human bronchial preparations. Immunohistochemical investigation and immunoradiometric assays with anti-M1 monoclonal antibodies (Mabs) were used to detect M1/MUC5AC mucin derived from bronchial preparations with an intact surface epithelium, or in tissues where the epithelium had been removed (rubbed preparations). The Mabs labelled both epithelial goblet cells and submucosal glandular cells in EGTA (4 mM)-exposed bronchial preparations, while only goblet cells were stained in EGTA (0.4 mM)-exposed tissues. The quantities of M1/MUC5AC mucin detected in either the bronchial fluids derived from EGTA (4 mM)-exposed intact and rubbed preparations or in bronchial fluids treated with EGTA (4 mM) were significantly increased by two-fold when compared with untreated control values (p<0.001). In addition, lactate dehydrogenase (LDH) activity and protein measurements were unaltered during exposure of human airways to EGTA (4 mM) suggesting that this treatment did not affect tissue viability. These results provide evidence that ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (4 mM) facilitates the detection of M1/MUC5AC mucin by altering the physicochemical properties of respiratory mucin, thereby exposing epitopes with which anti-M1 monoclonal antibodies are reactive. This will allow more accurate measurement of secretory activity in human airways in vitro.</description><subject>Biological and medical sciences</subject><subject>Bronchi - pathology</subject><subject>Egtazic Acid - pharmacology</subject><subject>Humans</subject><subject>In Vitro Techniques</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Lung Neoplasms - pathology</subject><subject>Medical sciences</subject><subject>Mucin 5AC</subject><subject>Mucins - analysis</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>Respiratory Mucosa - pathology</subject><subject>Respiratory system</subject><subject>Sensitivity and Specificity</subject><issn>0903-1936</issn><issn>1399-3003</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkFtPwzAMhSMEgnH5BwjlBd427KZpm8dp4iZt4oU9R2masEIvELcg_j2ZGCBZsiV_Pjo-jJ0jzBALcQ0KBCqRzQBnAAhSAO6xCQqlpgJA7LPJFplumSN2TPQSqSwVeMiOECVCrnDC1jd3T3M-BGeG1nUD7z3fjK3puKnDp_kiXnf8ox5Cz8euNfRKfIXXq_VCzhe8HW3cxqKxjHNPpuHPjekqOmUH3jTkznb9hK1vb54W99Pl493DYr6cWpGJIdr0BoVLXZECIpZpVRnhpZUycUokJZSQeCVd5TPlSkwK5ZPclmC9T8rEgThhVz-6b6F_Hx0Nuq3JuiaacP1IOkeELC9kBNMf0IaeKDiv30LdmvClEfQ2Tv0bpwbUv3HGs4udfvzQVf9Hu_wicLkDDFnT-GA6W9M_l0Imt7J_3KZ-3nzWwWlqTdNEWdQuvGChUWOeiW_5aoib</recordid><startdate>20010701</startdate><enddate>20010701</enddate><creator>Roger, P</creator><creator>Gascard, J-P</creator><creator>Bara, J</creator><creator>Dulmet, E</creator><creator>Brink, C</creator><general>Eur Respiratory Soc</general><general>Maney</general><scope>IQODW</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>7X8</scope></search><sort><creationdate>20010701</creationdate><title>EGTA treatment of human airways in vitro unmasks M1/MUC5AC mucin in submucosal glands</title><author>Roger, P ; Gascard, J-P ; Bara, J ; Dulmet, E ; Brink, C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c363t-30fa13e4e840111b4dda3f5c552e932b0b02f95edf69eb1289f27cb0cff2b2e03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Biological and medical sciences</topic><topic>Bronchi - pathology</topic><topic>Egtazic Acid - pharmacology</topic><topic>Humans</topic><topic>In Vitro Techniques</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Lung Neoplasms - pathology</topic><topic>Medical sciences</topic><topic>Mucin 5AC</topic><topic>Mucins - analysis</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>Respiratory Mucosa - pathology</topic><topic>Respiratory system</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Roger, P</creatorcontrib><creatorcontrib>Gascard, J-P</creatorcontrib><creatorcontrib>Bara, J</creatorcontrib><creatorcontrib>Dulmet, E</creatorcontrib><creatorcontrib>Brink, C</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The European respiratory journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Roger, P</au><au>Gascard, J-P</au><au>Bara, J</au><au>Dulmet, E</au><au>Brink, C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>EGTA treatment of human airways in vitro unmasks M1/MUC5AC mucin in submucosal glands</atitle><jtitle>The European respiratory journal</jtitle><addtitle>Eur Respir J</addtitle><date>2001-07-01</date><risdate>2001</risdate><volume>18</volume><issue>1</issue><spage>176</spage><epage>183</epage><pages>176-183</pages><issn>0903-1936</issn><eissn>1399-3003</eissn><abstract>Mucin staining can be used to evaluate secretory activity of human airways. However, mucin epitopes may be masked by physicochemical properties of the secretions. The aim of this investigation was to examine the effects of the calcium chelator, ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) on the detection of M1/MUC5AC mucin in isolated human bronchial preparations. Immunohistochemical investigation and immunoradiometric assays with anti-M1 monoclonal antibodies (Mabs) were used to detect M1/MUC5AC mucin derived from bronchial preparations with an intact surface epithelium, or in tissues where the epithelium had been removed (rubbed preparations). The Mabs labelled both epithelial goblet cells and submucosal glandular cells in EGTA (4 mM)-exposed bronchial preparations, while only goblet cells were stained in EGTA (0.4 mM)-exposed tissues. The quantities of M1/MUC5AC mucin detected in either the bronchial fluids derived from EGTA (4 mM)-exposed intact and rubbed preparations or in bronchial fluids treated with EGTA (4 mM) were significantly increased by two-fold when compared with untreated control values (p<0.001). In addition, lactate dehydrogenase (LDH) activity and protein measurements were unaltered during exposure of human airways to EGTA (4 mM) suggesting that this treatment did not affect tissue viability. These results provide evidence that ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (4 mM) facilitates the detection of M1/MUC5AC mucin by altering the physicochemical properties of respiratory mucin, thereby exposing epitopes with which anti-M1 monoclonal antibodies are reactive. This will allow more accurate measurement of secretory activity in human airways in vitro.</abstract><cop>Leeds</cop><pub>Eur Respiratory Soc</pub><pmid>11510791</pmid><doi>10.1183/09031936.01.00105301</doi><tpages>8</tpages></addata></record> |
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| subjects | Biological and medical sciences Bronchi - pathology Egtazic Acid - pharmacology Humans In Vitro Techniques Investigative techniques, diagnostic techniques (general aspects) Lung Neoplasms - pathology Medical sciences Mucin 5AC Mucins - analysis Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Respiratory Mucosa - pathology Respiratory system Sensitivity and Specificity |
| title | EGTA treatment of human airways in vitro unmasks M1/MUC5AC mucin in submucosal glands |
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