Whole-Blood Flow-Cytometric Analysis of Eosinophil EG2 Expression as a Marker of the Pathological Conditions of Asthma
Using a simple technique detecting the eosinophil fraction in whole-blood flow cytometry, we measured intracellular antigen EG2 (a monoclonal antibody to eosinophil cationic protein) in 56 asthmatic patients (26 during an attack and 30 during an asymptomatic period) and 22 healthy subjects to determ...
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| Veröffentlicht in: | International archives of allergy and immunology 1998-01, Vol.117 (Suppl 1), p.77-80 |
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| creator | Kuwasaki, Tomoe Chihara, Junichi Kayaba, Hiroyuki Kamata, Yumiko Oyamada, Hazime Saito, Norihiro Shioya, Takanobu Sasaki, Masahiro Kagaya, Manabu Tsuda, Akira |
| description | Using a simple technique detecting the eosinophil fraction in whole-blood flow cytometry, we measured intracellular antigen EG2 (a monoclonal antibody to eosinophil cationic protein) in 56 asthmatic patients (26 during an attack and 30 during an asymptomatic period) and 22 healthy subjects to determine whether EG2 reflects the pathological stages of allergy. Methods: In brief, preparations of the sample included the following procedures: (1) hemolyzation of heparinized or EDTA-mixed whole blood; (2) fixation of white blood cells with 0.4% parabenzoquinone (PBQ) or paraformaldehyde (PFA); (3) permeabilizing the cell membrane with n-octyl-β-D-glucopyranoside, and (4) staining of intracellular EG2 antigen with monoclonal EG2 antibody and FITC-labeled secondary antibody. Results: In PBQ-fixed samples, there was a clearer boundary of the eosinophil fraction with a higher yield and purity than in those fixed with PFA. The number of EG2-positive eosinophils was significantly greater in subjects during attacks than in asymptomatic patients. In addition, when compared with normal controls, asthmatic subjects had significantly greater numbers of EG2-positive eosinophils regardless of their current condition. Conclusion: Eosinophil intracellular EG2 may indicate the pathological stage of asthma. This simple technique for analysing the properties of eosinophils using whole-blood flow cytometry would save time and labor in laboratories. |
| doi_str_mv | 10.1159/000053578 |
| format | Article |
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Methods: In brief, preparations of the sample included the following procedures: (1) hemolyzation of heparinized or EDTA-mixed whole blood; (2) fixation of white blood cells with 0.4% parabenzoquinone (PBQ) or paraformaldehyde (PFA); (3) permeabilizing the cell membrane with n-octyl-β-D-glucopyranoside, and (4) staining of intracellular EG2 antigen with monoclonal EG2 antibody and FITC-labeled secondary antibody. Results: In PBQ-fixed samples, there was a clearer boundary of the eosinophil fraction with a higher yield and purity than in those fixed with PFA. The number of EG2-positive eosinophils was significantly greater in subjects during attacks than in asymptomatic patients. In addition, when compared with normal controls, asthmatic subjects had significantly greater numbers of EG2-positive eosinophils regardless of their current condition. Conclusion: Eosinophil intracellular EG2 may indicate the pathological stage of asthma. This simple technique for analysing the properties of eosinophils using whole-blood flow cytometry would save time and labor in laboratories.</description><identifier>ISSN: 1018-2438</identifier><identifier>ISBN: 9783805567923</identifier><identifier>ISBN: 3805567928</identifier><identifier>EISSN: 1423-0097</identifier><identifier>EISBN: 3318003700</identifier><identifier>EISBN: 9783318003703</identifier><identifier>DOI: 10.1159/000053578</identifier><identifier>PMID: 9758904</identifier><language>eng</language><publisher>Basel, Switzerland: Karger</publisher><subject>Adult ; Allergic diseases ; Asthma - blood ; Asthma - immunology ; Benzoquinones ; Biological and medical sciences ; Biomarkers - blood ; Blood Proteins - analysis ; Case-Control Studies ; Cell Membrane Permeability ; Eosinophil Granule Proteins ; Eosinophils - immunology ; Female ; Fixatives ; Flow Cytometry - methods ; Formaldehyde ; Glucosides ; Humans ; Immunopathology ; Inflammation Mediators - blood ; Male ; Medical sciences ; Middle Aged ; Polymers ; Respiratory and ent allergic diseases ; Ribonucleases</subject><ispartof>International archives of allergy and immunology, 1998-01, Vol.117 (Suppl 1), p.77-80</ispartof><rights>1998 S. Karger AG, Basel</rights><rights>1999 INIST-CNRS</rights><rights>Copyright (c) 1998 S. Karger AG, Basel</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c480t-76a23957658ea2192db793d89b7462e730738ce711d024c7b6946a27d43c7ca03</citedby><cites>FETCH-LOGICAL-c480t-76a23957658ea2192db793d89b7462e730738ce711d024c7b6946a27d43c7ca03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>310,311,315,781,785,790,791,2430,23935,23936,25145,27929,27930</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1582508$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9758904$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kuwasaki, Tomoe</creatorcontrib><creatorcontrib>Chihara, Junichi</creatorcontrib><creatorcontrib>Kayaba, Hiroyuki</creatorcontrib><creatorcontrib>Kamata, Yumiko</creatorcontrib><creatorcontrib>Oyamada, Hazime</creatorcontrib><creatorcontrib>Saito, Norihiro</creatorcontrib><creatorcontrib>Shioya, Takanobu</creatorcontrib><creatorcontrib>Sasaki, Masahiro</creatorcontrib><creatorcontrib>Kagaya, Manabu</creatorcontrib><creatorcontrib>Tsuda, Akira</creatorcontrib><title>Whole-Blood Flow-Cytometric Analysis of Eosinophil EG2 Expression as a Marker of the Pathological Conditions of Asthma</title><title>International archives of allergy and immunology</title><addtitle>Int Arch Allergy Immunol</addtitle><description>Using a simple technique detecting the eosinophil fraction in whole-blood flow cytometry, we measured intracellular antigen EG2 (a monoclonal antibody to eosinophil cationic protein) in 56 asthmatic patients (26 during an attack and 30 during an asymptomatic period) and 22 healthy subjects to determine whether EG2 reflects the pathological stages of allergy. Methods: In brief, preparations of the sample included the following procedures: (1) hemolyzation of heparinized or EDTA-mixed whole blood; (2) fixation of white blood cells with 0.4% parabenzoquinone (PBQ) or paraformaldehyde (PFA); (3) permeabilizing the cell membrane with n-octyl-β-D-glucopyranoside, and (4) staining of intracellular EG2 antigen with monoclonal EG2 antibody and FITC-labeled secondary antibody. Results: In PBQ-fixed samples, there was a clearer boundary of the eosinophil fraction with a higher yield and purity than in those fixed with PFA. The number of EG2-positive eosinophils was significantly greater in subjects during attacks than in asymptomatic patients. In addition, when compared with normal controls, asthmatic subjects had significantly greater numbers of EG2-positive eosinophils regardless of their current condition. Conclusion: Eosinophil intracellular EG2 may indicate the pathological stage of asthma. This simple technique for analysing the properties of eosinophils using whole-blood flow cytometry would save time and labor in laboratories.</description><subject>Adult</subject><subject>Allergic diseases</subject><subject>Asthma - blood</subject><subject>Asthma - immunology</subject><subject>Benzoquinones</subject><subject>Biological and medical sciences</subject><subject>Biomarkers - blood</subject><subject>Blood Proteins - analysis</subject><subject>Case-Control Studies</subject><subject>Cell Membrane Permeability</subject><subject>Eosinophil Granule Proteins</subject><subject>Eosinophils - immunology</subject><subject>Female</subject><subject>Fixatives</subject><subject>Flow Cytometry - methods</subject><subject>Formaldehyde</subject><subject>Glucosides</subject><subject>Humans</subject><subject>Immunopathology</subject><subject>Inflammation Mediators - blood</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Middle Aged</subject><subject>Polymers</subject><subject>Respiratory and ent allergic diseases</subject><subject>Ribonucleases</subject><issn>1018-2438</issn><issn>1423-0097</issn><isbn>9783805567923</isbn><isbn>3805567928</isbn><isbn>3318003700</isbn><isbn>9783318003703</isbn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkc1v0zAYh82XxlZ24IyQrAkhcQi8_ortY6i6MWkIDiCOkes4q7ckzuwU6H-Pu5YiISR88eH3-PGr94fQcwJvCRH6HeQjmJDqATphjCgAJgEeomPCKSsAtHyETrVUTIEQpdSUPc4ZEFVQztRTdJLSDUA2qfIIHWkplAZ-jL5_W4XOFe-7EBp83oUfxXwzhd5N0VtcDabbJJ9waPEiJD-EceU7vLigePFzjC4lHwZsEjb4o4m3Lm7BaeXwZzNlbbj21nR4HobGT5m891RpWvXmGXrSmi650_09Q1_PF1_mH4qrTxeX8-qqsFzBVMjSUKaFLIVyhhJNm6XUrFF6KXlJnWQgmbJOEtIA5VYuS83zE9lwZqU1wGbo9c47xnC3dmmqe5-s6zozuLBOtWSaU1my_4JEEqXKvMoZOvsLvAnrmBeVakqJysXkCWfozQ6yMaQUXVuP0fcmbmoC9bbO-lBnZl_uhetl75oDua8o56_2uUl5nW00g_Xpj1AoKmCrebHDbk28dvGQ__7k7J_pZVXdA_XYtOwXhve0RA</recordid><startdate>19980101</startdate><enddate>19980101</enddate><creator>Kuwasaki, Tomoe</creator><creator>Chihara, Junichi</creator><creator>Kayaba, Hiroyuki</creator><creator>Kamata, Yumiko</creator><creator>Oyamada, Hazime</creator><creator>Saito, Norihiro</creator><creator>Shioya, Takanobu</creator><creator>Sasaki, Masahiro</creator><creator>Kagaya, Manabu</creator><creator>Tsuda, Akira</creator><general>Karger</general><general>S. 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blood</topic><topic>Asthma - immunology</topic><topic>Benzoquinones</topic><topic>Biological and medical sciences</topic><topic>Biomarkers - blood</topic><topic>Blood Proteins - analysis</topic><topic>Case-Control Studies</topic><topic>Cell Membrane Permeability</topic><topic>Eosinophil Granule Proteins</topic><topic>Eosinophils - immunology</topic><topic>Female</topic><topic>Fixatives</topic><topic>Flow Cytometry - methods</topic><topic>Formaldehyde</topic><topic>Glucosides</topic><topic>Humans</topic><topic>Immunopathology</topic><topic>Inflammation Mediators - blood</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Middle Aged</topic><topic>Polymers</topic><topic>Respiratory and ent allergic diseases</topic><topic>Ribonucleases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kuwasaki, Tomoe</creatorcontrib><creatorcontrib>Chihara, Junichi</creatorcontrib><creatorcontrib>Kayaba, Hiroyuki</creatorcontrib><creatorcontrib>Kamata, Yumiko</creatorcontrib><creatorcontrib>Oyamada, Hazime</creatorcontrib><creatorcontrib>Saito, Norihiro</creatorcontrib><creatorcontrib>Shioya, Takanobu</creatorcontrib><creatorcontrib>Sasaki, Masahiro</creatorcontrib><creatorcontrib>Kagaya, Manabu</creatorcontrib><creatorcontrib>Tsuda, Akira</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>ProQuest Central (Corporate)</collection><collection>Immunology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Research Library</collection><collection>Biological Science Database</collection><collection>Research Library (Corporate)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><jtitle>International archives of allergy and immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kuwasaki, Tomoe</au><au>Chihara, Junichi</au><au>Kayaba, Hiroyuki</au><au>Kamata, Yumiko</au><au>Oyamada, Hazime</au><au>Saito, Norihiro</au><au>Shioya, Takanobu</au><au>Sasaki, Masahiro</au><au>Kagaya, Manabu</au><au>Tsuda, Akira</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Whole-Blood Flow-Cytometric Analysis of Eosinophil EG2 Expression as a Marker of the Pathological Conditions of Asthma</atitle><jtitle>International archives of allergy and immunology</jtitle><addtitle>Int Arch Allergy Immunol</addtitle><date>1998-01-01</date><risdate>1998</risdate><volume>117</volume><issue>Suppl 1</issue><spage>77</spage><epage>80</epage><pages>77-80</pages><issn>1018-2438</issn><eissn>1423-0097</eissn><isbn>9783805567923</isbn><isbn>3805567928</isbn><eisbn>3318003700</eisbn><eisbn>9783318003703</eisbn><abstract>Using a simple technique detecting the eosinophil fraction in whole-blood flow cytometry, we measured intracellular antigen EG2 (a monoclonal antibody to eosinophil cationic protein) in 56 asthmatic patients (26 during an attack and 30 during an asymptomatic period) and 22 healthy subjects to determine whether EG2 reflects the pathological stages of allergy. Methods: In brief, preparations of the sample included the following procedures: (1) hemolyzation of heparinized or EDTA-mixed whole blood; (2) fixation of white blood cells with 0.4% parabenzoquinone (PBQ) or paraformaldehyde (PFA); (3) permeabilizing the cell membrane with n-octyl-β-D-glucopyranoside, and (4) staining of intracellular EG2 antigen with monoclonal EG2 antibody and FITC-labeled secondary antibody. Results: In PBQ-fixed samples, there was a clearer boundary of the eosinophil fraction with a higher yield and purity than in those fixed with PFA. The number of EG2-positive eosinophils was significantly greater in subjects during attacks than in asymptomatic patients. In addition, when compared with normal controls, asthmatic subjects had significantly greater numbers of EG2-positive eosinophils regardless of their current condition. Conclusion: Eosinophil intracellular EG2 may indicate the pathological stage of asthma. This simple technique for analysing the properties of eosinophils using whole-blood flow cytometry would save time and labor in laboratories.</abstract><cop>Basel, Switzerland</cop><pub>Karger</pub><pmid>9758904</pmid><doi>10.1159/000053578</doi><tpages>4</tpages></addata></record> |
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| identifier | ISSN: 1018-2438 |
| ispartof | International archives of allergy and immunology, 1998-01, Vol.117 (Suppl 1), p.77-80 |
| issn | 1018-2438 1423-0097 |
| language | eng |
| recordid | cdi_pubmed_primary_9758904 |
| source | MEDLINE; Karger Journals; Alma/SFX Local Collection |
| subjects | Adult Allergic diseases Asthma - blood Asthma - immunology Benzoquinones Biological and medical sciences Biomarkers - blood Blood Proteins - analysis Case-Control Studies Cell Membrane Permeability Eosinophil Granule Proteins Eosinophils - immunology Female Fixatives Flow Cytometry - methods Formaldehyde Glucosides Humans Immunopathology Inflammation Mediators - blood Male Medical sciences Middle Aged Polymers Respiratory and ent allergic diseases Ribonucleases |
| title | Whole-Blood Flow-Cytometric Analysis of Eosinophil EG2 Expression as a Marker of the Pathological Conditions of Asthma |
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