Flow cytometric immunophenotypic analysis of systemic mastocytosis involving bone marrow
Mast cells of systemic mastocytosis (SM) have aberrant immunophenotypes that are useful for their detection by flow cytometry immunophenotyping. To assess the usefulness of CD2, CD25, and other antigens for establishing the diagnosis of SM in bone marrow using flow cytometry immunophenotyping. We st...
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| Veröffentlicht in: | Archives of pathology & laboratory medicine (1976) 2014-09, Vol.138 (9), p.1210-1214 |
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| creator | Jabbar, Kausar J Medeiros, L Jeffrey Wang, Sa A Miranda, Roberto N Johnson, Malisha R Verstovsek, Srdan Jorgensen, Jeffrey L |
| description | Mast cells of systemic mastocytosis (SM) have aberrant immunophenotypes that are useful for their detection by flow cytometry immunophenotyping.
To assess the usefulness of CD2, CD25, and other antigens for establishing the diagnosis of SM in bone marrow using flow cytometry immunophenotyping.
We studied 50 bone marrow aspirates of patients with SM using flow cytometry immunophenotyping. The bone marrow aspirates were stained with antibodies specific for CD2, CD25, CD35, CD59, CD63, and CD69. For the detection of CD2 and CD25, antibodies conjugated with phycoerythrin (PE) or fluorescein isothiocyanate (FITC) were compared. CD45-PerCP and CD117-APC were used for gating. Data were acquired on FACS Calibur cytometers and analyzed using CellQuest software.
CD2 and CD25 were positive in 41 of 50 (82%) and 45 of 50 (90%) SM cases, respectively. For CD2, the PE-conjugated antibody yielded better sensitivity than the FITC-conjugated antibody (31 of 40 [78%] versus 28 of 40 [70%]). For CD25, PE-conjugated and FITC-conjugated antibodies showed similar detection sensitivity, although the intensity of expression was brighter with CD25-PE. Compared with immunohistochemistry, flow cytometry immunophenotyping was superior for detecting CD2 (14 of 23 [61%] versus 9 of 23 [39%]). Other antigens frequently overexpressed in SM were CD35 (43 of 50 [86%]), CD59 (46 of 50 [92%]), CD63 (43 of 49 [88%]), and CD69 (39 of 48 [81%]).
Flow cytometry immunophenotyping is a rapid and sensitive technique for characterizing mast cells in bone marrow aspirate specimens. The use of PE-conjugated antibodies for CD2 and CD25 improves the detection rate (CD2) or facilitates analysis (CD25); therefore, PE-conjugated antibodies are suggested. Antibodies reactive with CD35, CD59, CD63, and CD69 are also helpful in detecting SM in bone marrow. |
| doi_str_mv | 10.5858/arpa.2013-0537-OA |
| format | Article |
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To assess the usefulness of CD2, CD25, and other antigens for establishing the diagnosis of SM in bone marrow using flow cytometry immunophenotyping.
We studied 50 bone marrow aspirates of patients with SM using flow cytometry immunophenotyping. The bone marrow aspirates were stained with antibodies specific for CD2, CD25, CD35, CD59, CD63, and CD69. For the detection of CD2 and CD25, antibodies conjugated with phycoerythrin (PE) or fluorescein isothiocyanate (FITC) were compared. CD45-PerCP and CD117-APC were used for gating. Data were acquired on FACS Calibur cytometers and analyzed using CellQuest software.
CD2 and CD25 were positive in 41 of 50 (82%) and 45 of 50 (90%) SM cases, respectively. For CD2, the PE-conjugated antibody yielded better sensitivity than the FITC-conjugated antibody (31 of 40 [78%] versus 28 of 40 [70%]). For CD25, PE-conjugated and FITC-conjugated antibodies showed similar detection sensitivity, although the intensity of expression was brighter with CD25-PE. Compared with immunohistochemistry, flow cytometry immunophenotyping was superior for detecting CD2 (14 of 23 [61%] versus 9 of 23 [39%]). Other antigens frequently overexpressed in SM were CD35 (43 of 50 [86%]), CD59 (46 of 50 [92%]), CD63 (43 of 49 [88%]), and CD69 (39 of 48 [81%]).
Flow cytometry immunophenotyping is a rapid and sensitive technique for characterizing mast cells in bone marrow aspirate specimens. The use of PE-conjugated antibodies for CD2 and CD25 improves the detection rate (CD2) or facilitates analysis (CD25); therefore, PE-conjugated antibodies are suggested. Antibodies reactive with CD35, CD59, CD63, and CD69 are also helpful in detecting SM in bone marrow.</description><identifier>ISSN: 0003-9985</identifier><identifier>ISSN: 1543-2165</identifier><identifier>EISSN: 1543-2165</identifier><identifier>DOI: 10.5858/arpa.2013-0537-OA</identifier><identifier>PMID: 25171703</identifier><identifier>CODEN: APLMAS</identifier><language>eng</language><publisher>United States: College of American Pathologists</publisher><subject>Adult ; Aged ; Aged, 80 and over ; Analysis ; Antibodies ; Antigens ; Antigens, CD - metabolism ; Antigens, Differentiation, T-Lymphocyte - metabolism ; Biopsy ; Blood diseases ; Bone marrow ; Bone Marrow - immunology ; Bone Marrow - pathology ; CD2 Antigens - metabolism ; CD59 Antigens - metabolism ; Cytoplasm ; Female ; Flow cytometry ; Flow Cytometry - methods ; Fluorescein ; Fluorescein-5-isothiocyanate ; Humans ; Immunohistochemistry ; Immunophenotyping - methods ; Interleukin-2 Receptor alpha Subunit - metabolism ; Lectins, C-Type - metabolism ; Leukemia ; Male ; Mast Cells - immunology ; Mast Cells - pathology ; Mastocytosis ; Mastocytosis, Systemic - diagnosis ; Mastocytosis, Systemic - immunology ; Mastocytosis, Systemic - pathology ; Medical research ; Middle Aged ; Patients ; Phycoerythrin ; Receptors, Complement 3b - metabolism ; Retrospective Studies ; Sensitivity and Specificity ; Software ; Tetraspanin 30 - metabolism</subject><ispartof>Archives of pathology & laboratory medicine (1976), 2014-09, Vol.138 (9), p.1210-1214</ispartof><rights>COPYRIGHT 2014 College of American Pathologists</rights><rights>Copyright College of American Pathologists Sep 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c403t-2c0780770ca6e9c2f17be98896d1399d80c25437daa4a8d793c8033ee6d6fbc43</citedby><cites>FETCH-LOGICAL-c403t-2c0780770ca6e9c2f17be98896d1399d80c25437daa4a8d793c8033ee6d6fbc43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25171703$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jabbar, Kausar J</creatorcontrib><creatorcontrib>Medeiros, L Jeffrey</creatorcontrib><creatorcontrib>Wang, Sa A</creatorcontrib><creatorcontrib>Miranda, Roberto N</creatorcontrib><creatorcontrib>Johnson, Malisha R</creatorcontrib><creatorcontrib>Verstovsek, Srdan</creatorcontrib><creatorcontrib>Jorgensen, Jeffrey L</creatorcontrib><title>Flow cytometric immunophenotypic analysis of systemic mastocytosis involving bone marrow</title><title>Archives of pathology & laboratory medicine (1976)</title><addtitle>Arch Pathol Lab Med</addtitle><description>Mast cells of systemic mastocytosis (SM) have aberrant immunophenotypes that are useful for their detection by flow cytometry immunophenotyping.
To assess the usefulness of CD2, CD25, and other antigens for establishing the diagnosis of SM in bone marrow using flow cytometry immunophenotyping.
We studied 50 bone marrow aspirates of patients with SM using flow cytometry immunophenotyping. The bone marrow aspirates were stained with antibodies specific for CD2, CD25, CD35, CD59, CD63, and CD69. For the detection of CD2 and CD25, antibodies conjugated with phycoerythrin (PE) or fluorescein isothiocyanate (FITC) were compared. CD45-PerCP and CD117-APC were used for gating. Data were acquired on FACS Calibur cytometers and analyzed using CellQuest software.
CD2 and CD25 were positive in 41 of 50 (82%) and 45 of 50 (90%) SM cases, respectively. For CD2, the PE-conjugated antibody yielded better sensitivity than the FITC-conjugated antibody (31 of 40 [78%] versus 28 of 40 [70%]). For CD25, PE-conjugated and FITC-conjugated antibodies showed similar detection sensitivity, although the intensity of expression was brighter with CD25-PE. Compared with immunohistochemistry, flow cytometry immunophenotyping was superior for detecting CD2 (14 of 23 [61%] versus 9 of 23 [39%]). Other antigens frequently overexpressed in SM were CD35 (43 of 50 [86%]), CD59 (46 of 50 [92%]), CD63 (43 of 49 [88%]), and CD69 (39 of 48 [81%]).
Flow cytometry immunophenotyping is a rapid and sensitive technique for characterizing mast cells in bone marrow aspirate specimens. The use of PE-conjugated antibodies for CD2 and CD25 improves the detection rate (CD2) or facilitates analysis (CD25); therefore, PE-conjugated antibodies are suggested. Antibodies reactive with CD35, CD59, CD63, and CD69 are also helpful in detecting SM in bone marrow.</description><subject>Adult</subject><subject>Aged</subject><subject>Aged, 80 and over</subject><subject>Analysis</subject><subject>Antibodies</subject><subject>Antigens</subject><subject>Antigens, CD - metabolism</subject><subject>Antigens, Differentiation, T-Lymphocyte - metabolism</subject><subject>Biopsy</subject><subject>Blood diseases</subject><subject>Bone marrow</subject><subject>Bone Marrow - immunology</subject><subject>Bone Marrow - pathology</subject><subject>CD2 Antigens - metabolism</subject><subject>CD59 Antigens - metabolism</subject><subject>Cytoplasm</subject><subject>Female</subject><subject>Flow cytometry</subject><subject>Flow Cytometry - methods</subject><subject>Fluorescein</subject><subject>Fluorescein-5-isothiocyanate</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Immunophenotyping - methods</subject><subject>Interleukin-2 Receptor alpha Subunit - metabolism</subject><subject>Lectins, C-Type - metabolism</subject><subject>Leukemia</subject><subject>Male</subject><subject>Mast Cells - immunology</subject><subject>Mast Cells - pathology</subject><subject>Mastocytosis</subject><subject>Mastocytosis, Systemic - diagnosis</subject><subject>Mastocytosis, Systemic - immunology</subject><subject>Mastocytosis, Systemic - pathology</subject><subject>Medical research</subject><subject>Middle Aged</subject><subject>Patients</subject><subject>Phycoerythrin</subject><subject>Receptors, Complement 3b - metabolism</subject><subject>Retrospective Studies</subject><subject>Sensitivity and Specificity</subject><subject>Software</subject><subject>Tetraspanin 30 - metabolism</subject><issn>0003-9985</issn><issn>1543-2165</issn><issn>1543-2165</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpdkUFrGzEQhUVpSVw3P6CXsFAovWyqkayVdDQmbgsGX1LoTchabbxBu9pIWpv999HiJIeexMx8T8ybh9BXwHdMMPFTh0HfEQy0xIzycr_-gBbAVrQkULGPaIExpqWUgl2jzzE-5VISAlfomjDgwDFdoH9b58-FmZLvbAqtKdquG3s_HG3v0zTkhu61m2IbC98UcYrJdrnZ6Zj8rJoHbX_y7tT2j8XB9zbPQvDnL-hTo120N6_vEv3d3j9sfpe7_a8_m_WuNCtMU0kM5gJzjo2urDSkAX6wUghZ1UClrAU2JDvitdYrLWouqRGYUmurumoOZkWX6Mfl3yH459HGpLo2Guuc7q0fowKWT0WAAM7ot__QJz-GbG-mKgICAPNMfb9Qj9pZdbTapWP0bkyt76NaMwAiKaczCBfQBB9jsI0aQpvNTwqwmvNRcz5qzkfN-aj9OmtuX1cYD52t3xVvgdAXVPeM1w</recordid><startdate>201409</startdate><enddate>201409</enddate><creator>Jabbar, Kausar J</creator><creator>Medeiros, L Jeffrey</creator><creator>Wang, Sa A</creator><creator>Miranda, Roberto N</creator><creator>Johnson, Malisha R</creator><creator>Verstovsek, Srdan</creator><creator>Jorgensen, Jeffrey L</creator><general>College of American Pathologists</general><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>3V.</scope><scope>4T-</scope><scope>4U-</scope><scope>7RV</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AF</scope><scope>8AO</scope><scope>8C1</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope></search><sort><creationdate>201409</creationdate><title>Flow cytometric immunophenotypic analysis of systemic mastocytosis involving bone marrow</title><author>Jabbar, Kausar J ; Medeiros, L Jeffrey ; Wang, Sa A ; Miranda, Roberto N ; Johnson, Malisha R ; Verstovsek, Srdan ; Jorgensen, Jeffrey L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c403t-2c0780770ca6e9c2f17be98896d1399d80c25437daa4a8d793c8033ee6d6fbc43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Adult</topic><topic>Aged</topic><topic>Aged, 80 and over</topic><topic>Analysis</topic><topic>Antibodies</topic><topic>Antigens</topic><topic>Antigens, CD - metabolism</topic><topic>Antigens, Differentiation, T-Lymphocyte - metabolism</topic><topic>Biopsy</topic><topic>Blood diseases</topic><topic>Bone marrow</topic><topic>Bone Marrow - immunology</topic><topic>Bone Marrow - pathology</topic><topic>CD2 Antigens - metabolism</topic><topic>CD59 Antigens - metabolism</topic><topic>Cytoplasm</topic><topic>Female</topic><topic>Flow cytometry</topic><topic>Flow Cytometry - methods</topic><topic>Fluorescein</topic><topic>Fluorescein-5-isothiocyanate</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Immunophenotyping - methods</topic><topic>Interleukin-2 Receptor alpha Subunit - metabolism</topic><topic>Lectins, C-Type - metabolism</topic><topic>Leukemia</topic><topic>Male</topic><topic>Mast Cells - immunology</topic><topic>Mast Cells - pathology</topic><topic>Mastocytosis</topic><topic>Mastocytosis, Systemic - diagnosis</topic><topic>Mastocytosis, Systemic - immunology</topic><topic>Mastocytosis, Systemic - pathology</topic><topic>Medical research</topic><topic>Middle Aged</topic><topic>Patients</topic><topic>Phycoerythrin</topic><topic>Receptors, Complement 3b - metabolism</topic><topic>Retrospective Studies</topic><topic>Sensitivity and Specificity</topic><topic>Software</topic><topic>Tetraspanin 30 - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jabbar, Kausar J</creatorcontrib><creatorcontrib>Medeiros, L Jeffrey</creatorcontrib><creatorcontrib>Wang, Sa A</creatorcontrib><creatorcontrib>Miranda, Roberto N</creatorcontrib><creatorcontrib>Johnson, Malisha R</creatorcontrib><creatorcontrib>Verstovsek, Srdan</creatorcontrib><creatorcontrib>Jorgensen, Jeffrey L</creatorcontrib><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>Docstoc</collection><collection>University Readers</collection><collection>Nursing & Allied Health Database</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>STEM Database</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</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>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>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Nursing & Allied Health Premium</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>Archives of pathology & laboratory medicine (1976)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jabbar, Kausar J</au><au>Medeiros, L Jeffrey</au><au>Wang, Sa A</au><au>Miranda, Roberto N</au><au>Johnson, Malisha R</au><au>Verstovsek, Srdan</au><au>Jorgensen, Jeffrey L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Flow cytometric immunophenotypic analysis of systemic mastocytosis involving bone marrow</atitle><jtitle>Archives of pathology & laboratory medicine (1976)</jtitle><addtitle>Arch Pathol Lab Med</addtitle><date>2014-09</date><risdate>2014</risdate><volume>138</volume><issue>9</issue><spage>1210</spage><epage>1214</epage><pages>1210-1214</pages><issn>0003-9985</issn><issn>1543-2165</issn><eissn>1543-2165</eissn><coden>APLMAS</coden><abstract>Mast cells of systemic mastocytosis (SM) have aberrant immunophenotypes that are useful for their detection by flow cytometry immunophenotyping.
To assess the usefulness of CD2, CD25, and other antigens for establishing the diagnosis of SM in bone marrow using flow cytometry immunophenotyping.
We studied 50 bone marrow aspirates of patients with SM using flow cytometry immunophenotyping. The bone marrow aspirates were stained with antibodies specific for CD2, CD25, CD35, CD59, CD63, and CD69. For the detection of CD2 and CD25, antibodies conjugated with phycoerythrin (PE) or fluorescein isothiocyanate (FITC) were compared. CD45-PerCP and CD117-APC were used for gating. Data were acquired on FACS Calibur cytometers and analyzed using CellQuest software.
CD2 and CD25 were positive in 41 of 50 (82%) and 45 of 50 (90%) SM cases, respectively. For CD2, the PE-conjugated antibody yielded better sensitivity than the FITC-conjugated antibody (31 of 40 [78%] versus 28 of 40 [70%]). For CD25, PE-conjugated and FITC-conjugated antibodies showed similar detection sensitivity, although the intensity of expression was brighter with CD25-PE. Compared with immunohistochemistry, flow cytometry immunophenotyping was superior for detecting CD2 (14 of 23 [61%] versus 9 of 23 [39%]). Other antigens frequently overexpressed in SM were CD35 (43 of 50 [86%]), CD59 (46 of 50 [92%]), CD63 (43 of 49 [88%]), and CD69 (39 of 48 [81%]).
Flow cytometry immunophenotyping is a rapid and sensitive technique for characterizing mast cells in bone marrow aspirate specimens. The use of PE-conjugated antibodies for CD2 and CD25 improves the detection rate (CD2) or facilitates analysis (CD25); therefore, PE-conjugated antibodies are suggested. Antibodies reactive with CD35, CD59, CD63, and CD69 are also helpful in detecting SM in bone marrow.</abstract><cop>United States</cop><pub>College of American Pathologists</pub><pmid>25171703</pmid><doi>10.5858/arpa.2013-0537-OA</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adult Aged Aged, 80 and over Analysis Antibodies Antigens Antigens, CD - metabolism Antigens, Differentiation, T-Lymphocyte - metabolism Biopsy Blood diseases Bone marrow Bone Marrow - immunology Bone Marrow - pathology CD2 Antigens - metabolism CD59 Antigens - metabolism Cytoplasm Female Flow cytometry Flow Cytometry - methods Fluorescein Fluorescein-5-isothiocyanate Humans Immunohistochemistry Immunophenotyping - methods Interleukin-2 Receptor alpha Subunit - metabolism Lectins, C-Type - metabolism Leukemia Male Mast Cells - immunology Mast Cells - pathology Mastocytosis Mastocytosis, Systemic - diagnosis Mastocytosis, Systemic - immunology Mastocytosis, Systemic - pathology Medical research Middle Aged Patients Phycoerythrin Receptors, Complement 3b - metabolism Retrospective Studies Sensitivity and Specificity Software Tetraspanin 30 - metabolism |
| title | Flow cytometric immunophenotypic analysis of systemic mastocytosis involving bone marrow |
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