Sputum processing for evaluation of inflammatory mediators
Neutrophil‐dominated inflammation is prominent in the cystic fibrosis (CF) and chronic bronchitis (CB) airways. We assessed the degree of airway inflammation by measuring the sputum concentrations of interleukin (IL)‐8, myeloperoxidase (MPO), and deoxyribonucleic acid (DNA). We determined the relati...
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| Veröffentlicht in: | Pediatric pulmonology 2001-08, Vol.32 (2), p.152-158 |
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| creator | Kim, Jung-Soo Hackley, Grant H. Okamoto, Kosuke Rubin, Bruce K. |
| description | Neutrophil‐dominated inflammation is prominent in the cystic fibrosis (CF) and chronic bronchitis (CB) airways. We assessed the degree of airway inflammation by measuring the sputum concentrations of interleukin (IL)‐8, myeloperoxidase (MPO), and deoxyribonucleic acid (DNA). We determined the relationship among the concentrations of these mediators and investigated methodological problems that may be responsible for reported variability in measurements. Sputa obtained from 31 patients were solubilized with phosphate‐buffered saline, dithiothreitol (DTT) (0.1% or 1%), or dornase alfa (0.2 mg/mL). The sputum concentration of IL‐8 and MPO was measured by enzyme‐linked immunosorbent assay (ELISA), and DNA was measured using microfluorimetry.
There was a significant relationship among sputum IL‐8, MPO, and DNA. For MPO (means ± SD), CF was 1,392 ± 771 vs. CB at 75 ± 65 mcg/mL; P |
| doi_str_mv | 10.1002/ppul.1101 |
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There was a significant relationship among sputum IL‐8, MPO, and DNA. For MPO (means ± SD), CF was 1,392 ± 771 vs. CB at 75 ± 65 mcg/mL; P < 0.0001. For IL‐8: CF was 239 ± 154 vs. CB at 121 ± 108 ng/mL; P = 0.0002. For DNA, CF was 1.707 ± 1.25 vs. CB at 0.184 ± 0.272 mg/mL; P < 0.0001. The MPO concentration in CF sputum was approximately double after in vitro treatment with dornase alfa (P < 0.0001).
There is a greater concentration of IL‐8, MPO, and DNA in CF than in CB sputa. There is a significant relationship among these inflammatory markers in sputum. DNA polymers bind myeloperoxidase in the sputum, and we speculate that treatment with dornase alfa may remove a source of MPO inhibition. Pediatr Pulmonol. 2001; 32:152–158. © 2001 Wiley‐Liss, Inc.</description><identifier>ISSN: 8755-6863</identifier><identifier>EISSN: 1099-0496</identifier><identifier>DOI: 10.1002/ppul.1101</identifier><identifier>PMID: 11477732</identifier><identifier>CODEN: PEPUES</identifier><language>eng</language><publisher>New York: John Wiley & Sons, Inc</publisher><subject>Adolescent ; Adult ; Biological and medical sciences ; Biomarkers - analysis ; Bronchitis - physiopathology ; Child ; chronic bronchitis ; cystic fibrosis ; Cystic Fibrosis - physiopathology ; Deoxyribonuclease I - pharmacology ; dithiothreitol ; DNA ; DNA - analysis ; dornase alfa ; Expectorants - pharmacology ; Female ; Humans ; Inflammation ; interleukin-8 ; Interleukin-8 - analysis ; Investigative techniques, diagnostic techniques (general aspects) ; Male ; Medical sciences ; myeloperoxidase ; neutrophils ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Peroxidase - analysis ; Polymers ; Recombinant Proteins - pharmacology ; Respiratory system ; Specimen Handling ; Sputum - chemistry</subject><ispartof>Pediatric pulmonology, 2001-08, Vol.32 (2), p.152-158</ispartof><rights>Copyright © 2001 Wiley‐Liss, Inc.</rights><rights>2001 INIST-CNRS</rights><rights>Copyright 2001 Wiley-Liss, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3031-7e19fe8d5c016aa6a079bca1df38cdd7b1df878067d5350090d804af96f4a5ef3</citedby><cites>FETCH-LOGICAL-c3031-7e19fe8d5c016aa6a079bca1df38cdd7b1df878067d5350090d804af96f4a5ef3</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%2Fppul.1101$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fppul.1101$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1083038$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11477732$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, Jung-Soo</creatorcontrib><creatorcontrib>Hackley, Grant H.</creatorcontrib><creatorcontrib>Okamoto, Kosuke</creatorcontrib><creatorcontrib>Rubin, Bruce K.</creatorcontrib><title>Sputum processing for evaluation of inflammatory mediators</title><title>Pediatric pulmonology</title><addtitle>Pediatr. Pulmonol</addtitle><description>Neutrophil‐dominated inflammation is prominent in the cystic fibrosis (CF) and chronic bronchitis (CB) airways. We assessed the degree of airway inflammation by measuring the sputum concentrations of interleukin (IL)‐8, myeloperoxidase (MPO), and deoxyribonucleic acid (DNA). We determined the relationship among the concentrations of these mediators and investigated methodological problems that may be responsible for reported variability in measurements. Sputa obtained from 31 patients were solubilized with phosphate‐buffered saline, dithiothreitol (DTT) (0.1% or 1%), or dornase alfa (0.2 mg/mL). The sputum concentration of IL‐8 and MPO was measured by enzyme‐linked immunosorbent assay (ELISA), and DNA was measured using microfluorimetry.
There was a significant relationship among sputum IL‐8, MPO, and DNA. For MPO (means ± SD), CF was 1,392 ± 771 vs. CB at 75 ± 65 mcg/mL; P < 0.0001. For IL‐8: CF was 239 ± 154 vs. CB at 121 ± 108 ng/mL; P = 0.0002. For DNA, CF was 1.707 ± 1.25 vs. CB at 0.184 ± 0.272 mg/mL; P < 0.0001. The MPO concentration in CF sputum was approximately double after in vitro treatment with dornase alfa (P < 0.0001).
There is a greater concentration of IL‐8, MPO, and DNA in CF than in CB sputa. There is a significant relationship among these inflammatory markers in sputum. DNA polymers bind myeloperoxidase in the sputum, and we speculate that treatment with dornase alfa may remove a source of MPO inhibition. Pediatr Pulmonol. 2001; 32:152–158. © 2001 Wiley‐Liss, Inc.</description><subject>Adolescent</subject><subject>Adult</subject><subject>Biological and medical sciences</subject><subject>Biomarkers - analysis</subject><subject>Bronchitis - physiopathology</subject><subject>Child</subject><subject>chronic bronchitis</subject><subject>cystic fibrosis</subject><subject>Cystic Fibrosis - physiopathology</subject><subject>Deoxyribonuclease I - pharmacology</subject><subject>dithiothreitol</subject><subject>DNA</subject><subject>DNA - analysis</subject><subject>dornase alfa</subject><subject>Expectorants - pharmacology</subject><subject>Female</subject><subject>Humans</subject><subject>Inflammation</subject><subject>interleukin-8</subject><subject>Interleukin-8 - analysis</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Male</subject><subject>Medical sciences</subject><subject>myeloperoxidase</subject><subject>neutrophils</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>Peroxidase - analysis</subject><subject>Polymers</subject><subject>Recombinant Proteins - pharmacology</subject><subject>Respiratory system</subject><subject>Specimen Handling</subject><subject>Sputum - chemistry</subject><issn>8755-6863</issn><issn>1099-0496</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kEtP3TAQha2qqFweC_4AygJV6iIwg5PYYYdQeaiXhwSoFRtrrmNXbvOqnRTuv8dXN6JsupqR5pszZw5jewiHCHB81PdjfYgI-IHNEMoyhawsPrKZFHmeFrLgm2wrhF8AcVbiJ7aJmAkh-PGMndz34zA2Se87bUJw7c_Edj4xf6keaXBdm3Q2ca2tqWlo6PwyaUzlVl3YYRuW6mB2p7rNHs-_PpxdpvPbi6uz03mqOXBMhcHSGlnlGrAgKghEudCEleVSV5VYxE4KCYWocp5Hi1BJyMiWhc0oN5Zvs89r3ejxz2jCoBoXtKlrak03BiUQcuQZRPDLGtS-C8Ebq3rvGvJLhaBWQalVUGoVVGT3J9FxET_6R07JROBgAihoqq2nVrvwTlHG72TEjtbYs6vN8v8H1d3d43y6nK43XBjMy9sG-d-qEFzk6vvNhXq6_jH_Vj6dK-Svx2SPYg</recordid><startdate>200108</startdate><enddate>200108</enddate><creator>Kim, Jung-Soo</creator><creator>Hackley, Grant H.</creator><creator>Okamoto, Kosuke</creator><creator>Rubin, Bruce K.</creator><general>John Wiley & Sons, Inc</general><general>Wiley-Liss</general><scope>BSCLL</scope><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>200108</creationdate><title>Sputum processing for evaluation of inflammatory mediators</title><author>Kim, Jung-Soo ; Hackley, Grant H. ; Okamoto, Kosuke ; Rubin, Bruce K.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3031-7e19fe8d5c016aa6a079bca1df38cdd7b1df878067d5350090d804af96f4a5ef3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Adolescent</topic><topic>Adult</topic><topic>Biological and medical sciences</topic><topic>Biomarkers - analysis</topic><topic>Bronchitis - physiopathology</topic><topic>Child</topic><topic>chronic bronchitis</topic><topic>cystic fibrosis</topic><topic>Cystic Fibrosis - physiopathology</topic><topic>Deoxyribonuclease I - pharmacology</topic><topic>dithiothreitol</topic><topic>DNA</topic><topic>DNA - analysis</topic><topic>dornase alfa</topic><topic>Expectorants - pharmacology</topic><topic>Female</topic><topic>Humans</topic><topic>Inflammation</topic><topic>interleukin-8</topic><topic>Interleukin-8 - analysis</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Male</topic><topic>Medical sciences</topic><topic>myeloperoxidase</topic><topic>neutrophils</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>Peroxidase - analysis</topic><topic>Polymers</topic><topic>Recombinant Proteins - pharmacology</topic><topic>Respiratory system</topic><topic>Specimen Handling</topic><topic>Sputum - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Jung-Soo</creatorcontrib><creatorcontrib>Hackley, Grant H.</creatorcontrib><creatorcontrib>Okamoto, Kosuke</creatorcontrib><creatorcontrib>Rubin, Bruce K.</creatorcontrib><collection>Istex</collection><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>Pediatric pulmonology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Jung-Soo</au><au>Hackley, Grant H.</au><au>Okamoto, Kosuke</au><au>Rubin, Bruce K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sputum processing for evaluation of inflammatory mediators</atitle><jtitle>Pediatric pulmonology</jtitle><addtitle>Pediatr. Pulmonol</addtitle><date>2001-08</date><risdate>2001</risdate><volume>32</volume><issue>2</issue><spage>152</spage><epage>158</epage><pages>152-158</pages><issn>8755-6863</issn><eissn>1099-0496</eissn><coden>PEPUES</coden><abstract>Neutrophil‐dominated inflammation is prominent in the cystic fibrosis (CF) and chronic bronchitis (CB) airways. We assessed the degree of airway inflammation by measuring the sputum concentrations of interleukin (IL)‐8, myeloperoxidase (MPO), and deoxyribonucleic acid (DNA). We determined the relationship among the concentrations of these mediators and investigated methodological problems that may be responsible for reported variability in measurements. Sputa obtained from 31 patients were solubilized with phosphate‐buffered saline, dithiothreitol (DTT) (0.1% or 1%), or dornase alfa (0.2 mg/mL). The sputum concentration of IL‐8 and MPO was measured by enzyme‐linked immunosorbent assay (ELISA), and DNA was measured using microfluorimetry.
There was a significant relationship among sputum IL‐8, MPO, and DNA. For MPO (means ± SD), CF was 1,392 ± 771 vs. CB at 75 ± 65 mcg/mL; P < 0.0001. For IL‐8: CF was 239 ± 154 vs. CB at 121 ± 108 ng/mL; P = 0.0002. For DNA, CF was 1.707 ± 1.25 vs. CB at 0.184 ± 0.272 mg/mL; P < 0.0001. The MPO concentration in CF sputum was approximately double after in vitro treatment with dornase alfa (P < 0.0001).
There is a greater concentration of IL‐8, MPO, and DNA in CF than in CB sputa. There is a significant relationship among these inflammatory markers in sputum. DNA polymers bind myeloperoxidase in the sputum, and we speculate that treatment with dornase alfa may remove a source of MPO inhibition. Pediatr Pulmonol. 2001; 32:152–158. © 2001 Wiley‐Liss, Inc.</abstract><cop>New York</cop><pub>John Wiley & Sons, Inc</pub><pmid>11477732</pmid><doi>10.1002/ppul.1101</doi><tpages>7</tpages></addata></record> |
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| subjects | Adolescent Adult Biological and medical sciences Biomarkers - analysis Bronchitis - physiopathology Child chronic bronchitis cystic fibrosis Cystic Fibrosis - physiopathology Deoxyribonuclease I - pharmacology dithiothreitol DNA DNA - analysis dornase alfa Expectorants - pharmacology Female Humans Inflammation interleukin-8 Interleukin-8 - analysis Investigative techniques, diagnostic techniques (general aspects) Male Medical sciences myeloperoxidase neutrophils Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Peroxidase - analysis Polymers Recombinant Proteins - pharmacology Respiratory system Specimen Handling Sputum - chemistry |
| title | Sputum processing for evaluation of inflammatory mediators |
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