Surface Molecule Loss and Bleb Formation by Human Germinal Center B Cells Undergoing Apoptosis: Role of Apoptotic Blebs in Monocyte Chemotaxis
Human tonsil germinal center (GC) B cells rapidly undergo apoptosis in culture. Annexin-V binding shows an early event in this process. In the present study, this method has been used to label apoptotic GC B cells and to analyze additional surface molecules. The expression of all of the molecules st...
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Veröffentlicht in: | Blood 1999-08, Vol.94 (3), p.1012-1020 |
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description | Human tonsil germinal center (GC) B cells rapidly undergo apoptosis in culture. Annexin-V binding shows an early event in this process. In the present study, this method has been used to label apoptotic GC B cells and to analyze additional surface molecules. The expression of all of the molecules studied was reduced in apoptotic (annexin-V+) GC B cells, and the reduction was more marked for CD11a, CD21, CD22, CD49d, and CD54, molecules that participate in survival interaction for GC B cells. The analysis of CD54, one of the molecules that was more drastically reduced, showed that GC, but not mantle zone, B cells actively secrete CD54 to the culture supernatant (SN). The secreted CD54 was partly released from the GC B cells in a particulate form as demonstrated by centrifugation. Further experiments using filtration, fluorescence microscopy, electron microscopy, and flow cytometry analysis showed that GC B cells released to the culture SN a population of spherical membranous vesicles of about 0.18 μm in size, similar to the blebs described in other apoptosis systems. Bleb formation depended on active metabolism, Ca2+, and, in part, on microfilament integrity. GC B-cell–derived blebs were clearly associated with apoptosis, as antiapoptotic stimuli prevented their formation. In addition, GC B-cell–derived blebs contained the adhesion molecules previously studied. Consequently, bleb formation might contribute to the surface molecule loss occurring in apoptotic GC B cells. Finally, a chemotaxis assay showed that GC B-cell blebs were chemotactic for human monocytes, suggesting that this mechanism might operate in vivo. |
doi_str_mv | 10.1182/blood.V94.3.1012.415k05_1012_1020 |
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Annexin-V binding shows an early event in this process. In the present study, this method has been used to label apoptotic GC B cells and to analyze additional surface molecules. The expression of all of the molecules studied was reduced in apoptotic (annexin-V+) GC B cells, and the reduction was more marked for CD11a, CD21, CD22, CD49d, and CD54, molecules that participate in survival interaction for GC B cells. The analysis of CD54, one of the molecules that was more drastically reduced, showed that GC, but not mantle zone, B cells actively secrete CD54 to the culture supernatant (SN). The secreted CD54 was partly released from the GC B cells in a particulate form as demonstrated by centrifugation. Further experiments using filtration, fluorescence microscopy, electron microscopy, and flow cytometry analysis showed that GC B cells released to the culture SN a population of spherical membranous vesicles of about 0.18 μm in size, similar to the blebs described in other apoptosis systems. Bleb formation depended on active metabolism, Ca2+, and, in part, on microfilament integrity. GC B-cell–derived blebs were clearly associated with apoptosis, as antiapoptotic stimuli prevented their formation. In addition, GC B-cell–derived blebs contained the adhesion molecules previously studied. Consequently, bleb formation might contribute to the surface molecule loss occurring in apoptotic GC B cells. Finally, a chemotaxis assay showed that GC B-cell blebs were chemotactic for human monocytes, suggesting that this mechanism might operate in vivo.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood.V94.3.1012.415k05_1012_1020</identifier><identifier>PMID: 10419893</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Ageing, cell death ; Antigens, CD - immunology ; Apoptosis - immunology ; B-Lymphocytes - immunology ; B-Lymphocytes - pathology ; Biological and medical sciences ; Cell Adhesion Molecules - immunology ; Cell physiology ; Cells, Cultured ; Chemotaxis, Leukocyte - immunology ; Cytoplasmic Granules - immunology ; Fundamental and applied biological sciences. Psychology ; Humans ; Molecular and cellular biology ; Monocytes - immunology ; Monocytes - pathology</subject><ispartof>Blood, 1999-08, Vol.94 (3), p.1012-1020</ispartof><rights>1999 American Society of Hematology</rights><rights>1999 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c556t-aabde4b7f343f952829db5bf675fddb73db5b1dc9688a43e470eccd2e880bd9b3</citedby><cites>FETCH-LOGICAL-c556t-aabde4b7f343f952829db5bf675fddb73db5b1dc9688a43e470eccd2e880bd9b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1912347$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10419893$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Segundo, Carmen</creatorcontrib><creatorcontrib>Medina, Francisco</creatorcontrib><creatorcontrib>Rodrı́guez, Carmen</creatorcontrib><creatorcontrib>Martı́nez-Palencia, Rosalı́a</creatorcontrib><creatorcontrib>Leyva-Cobián, Francisco</creatorcontrib><creatorcontrib>Brieva, José A.</creatorcontrib><title>Surface Molecule Loss and Bleb Formation by Human Germinal Center B Cells Undergoing Apoptosis: Role of Apoptotic Blebs in Monocyte Chemotaxis</title><title>Blood</title><addtitle>Blood</addtitle><description>Human tonsil germinal center (GC) B cells rapidly undergo apoptosis in culture. Annexin-V binding shows an early event in this process. In the present study, this method has been used to label apoptotic GC B cells and to analyze additional surface molecules. The expression of all of the molecules studied was reduced in apoptotic (annexin-V+) GC B cells, and the reduction was more marked for CD11a, CD21, CD22, CD49d, and CD54, molecules that participate in survival interaction for GC B cells. The analysis of CD54, one of the molecules that was more drastically reduced, showed that GC, but not mantle zone, B cells actively secrete CD54 to the culture supernatant (SN). The secreted CD54 was partly released from the GC B cells in a particulate form as demonstrated by centrifugation. Further experiments using filtration, fluorescence microscopy, electron microscopy, and flow cytometry analysis showed that GC B cells released to the culture SN a population of spherical membranous vesicles of about 0.18 μm in size, similar to the blebs described in other apoptosis systems. Bleb formation depended on active metabolism, Ca2+, and, in part, on microfilament integrity. GC B-cell–derived blebs were clearly associated with apoptosis, as antiapoptotic stimuli prevented their formation. In addition, GC B-cell–derived blebs contained the adhesion molecules previously studied. Consequently, bleb formation might contribute to the surface molecule loss occurring in apoptotic GC B cells. Finally, a chemotaxis assay showed that GC B-cell blebs were chemotactic for human monocytes, suggesting that this mechanism might operate in vivo.</description><subject>Ageing, cell death</subject><subject>Antigens, CD - immunology</subject><subject>Apoptosis - immunology</subject><subject>B-Lymphocytes - immunology</subject><subject>B-Lymphocytes - pathology</subject><subject>Biological and medical sciences</subject><subject>Cell Adhesion Molecules - immunology</subject><subject>Cell physiology</subject><subject>Cells, Cultured</subject><subject>Chemotaxis, Leukocyte - immunology</subject><subject>Cytoplasmic Granules - immunology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Molecular and cellular biology</subject><subject>Monocytes - immunology</subject><subject>Monocytes - pathology</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkctu1DAUhi0EokPhFZAXCIlFgh07F7ObjnpBDEICytby5aQYEnuwk4p5CZ4ZTzMSFbtu7GPr03_O-X-E3lBSUtpVb_UQgi2_CV6ykhJalZzWP0ktD3U-KvIIrWhddQXJ9WO0IoQ0BRctPUHPUvpBCOWsqp-iE0o4FZ1gK_Tnyxx7ZQB_DAOYeQC8DSlh5S0-G0DjixBHNbngsd7jq3lUHl9CHJ1XA96AnyDis1wMQ8LX3kK8Cc7f4PUu7KaQXHqHP2ddHPrj1-TMnW7CzueWPpj9BHjzHcYwqd8uPUdPejUkeHG8T9H1xfnXzVWx_XT5frPeFqaum6lQSlvguu0ZZ73IG1fC6lr3TVv31uqWHV7UGtF0neIMeEvAGFtB1xFthWan6PWiu4vh1wxpkqNLJq-hPIQ5yUYIWjWcZHC9gCZmXyL0chfdqOI--y0Poci7UGQORbIliP9DyRovj81mPYK9p7CkkIFXR0Alo4Y-Km9c-sflWRhvM_ZhwSA7c-sgymQceAPWRTCTtME9YKq_dZWy0g</recordid><startdate>19990801</startdate><enddate>19990801</enddate><creator>Segundo, Carmen</creator><creator>Medina, Francisco</creator><creator>Rodrı́guez, Carmen</creator><creator>Martı́nez-Palencia, Rosalı́a</creator><creator>Leyva-Cobián, Francisco</creator><creator>Brieva, José A.</creator><general>Elsevier Inc</general><general>The Americain Society of Hematology</general><scope>6I.</scope><scope>AAFTH</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>19990801</creationdate><title>Surface Molecule Loss and Bleb Formation by Human Germinal Center B Cells Undergoing Apoptosis: Role of Apoptotic Blebs in Monocyte Chemotaxis</title><author>Segundo, Carmen ; Medina, Francisco ; Rodrı́guez, Carmen ; Martı́nez-Palencia, Rosalı́a ; Leyva-Cobián, Francisco ; Brieva, José A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c556t-aabde4b7f343f952829db5bf675fddb73db5b1dc9688a43e470eccd2e880bd9b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Ageing, cell death</topic><topic>Antigens, CD - immunology</topic><topic>Apoptosis - immunology</topic><topic>B-Lymphocytes - immunology</topic><topic>B-Lymphocytes - pathology</topic><topic>Biological and medical sciences</topic><topic>Cell Adhesion Molecules - immunology</topic><topic>Cell physiology</topic><topic>Cells, Cultured</topic><topic>Chemotaxis, Leukocyte - immunology</topic><topic>Cytoplasmic Granules - immunology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Molecular and cellular biology</topic><topic>Monocytes - immunology</topic><topic>Monocytes - pathology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Segundo, Carmen</creatorcontrib><creatorcontrib>Medina, Francisco</creatorcontrib><creatorcontrib>Rodrı́guez, Carmen</creatorcontrib><creatorcontrib>Martı́nez-Palencia, Rosalı́a</creatorcontrib><creatorcontrib>Leyva-Cobián, Francisco</creatorcontrib><creatorcontrib>Brieva, José A.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</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>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Segundo, Carmen</au><au>Medina, Francisco</au><au>Rodrı́guez, Carmen</au><au>Martı́nez-Palencia, Rosalı́a</au><au>Leyva-Cobián, Francisco</au><au>Brieva, José A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Surface Molecule Loss and Bleb Formation by Human Germinal Center B Cells Undergoing Apoptosis: Role of Apoptotic Blebs in Monocyte Chemotaxis</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>1999-08-01</date><risdate>1999</risdate><volume>94</volume><issue>3</issue><spage>1012</spage><epage>1020</epage><pages>1012-1020</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><abstract>Human tonsil germinal center (GC) B cells rapidly undergo apoptosis in culture. Annexin-V binding shows an early event in this process. In the present study, this method has been used to label apoptotic GC B cells and to analyze additional surface molecules. The expression of all of the molecules studied was reduced in apoptotic (annexin-V+) GC B cells, and the reduction was more marked for CD11a, CD21, CD22, CD49d, and CD54, molecules that participate in survival interaction for GC B cells. The analysis of CD54, one of the molecules that was more drastically reduced, showed that GC, but not mantle zone, B cells actively secrete CD54 to the culture supernatant (SN). The secreted CD54 was partly released from the GC B cells in a particulate form as demonstrated by centrifugation. Further experiments using filtration, fluorescence microscopy, electron microscopy, and flow cytometry analysis showed that GC B cells released to the culture SN a population of spherical membranous vesicles of about 0.18 μm in size, similar to the blebs described in other apoptosis systems. Bleb formation depended on active metabolism, Ca2+, and, in part, on microfilament integrity. GC B-cell–derived blebs were clearly associated with apoptosis, as antiapoptotic stimuli prevented their formation. In addition, GC B-cell–derived blebs contained the adhesion molecules previously studied. Consequently, bleb formation might contribute to the surface molecule loss occurring in apoptotic GC B cells. Finally, a chemotaxis assay showed that GC B-cell blebs were chemotactic for human monocytes, suggesting that this mechanism might operate in vivo.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>10419893</pmid><doi>10.1182/blood.V94.3.1012.415k05_1012_1020</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Ageing, cell death Antigens, CD - immunology Apoptosis - immunology B-Lymphocytes - immunology B-Lymphocytes - pathology Biological and medical sciences Cell Adhesion Molecules - immunology Cell physiology Cells, Cultured Chemotaxis, Leukocyte - immunology Cytoplasmic Granules - immunology Fundamental and applied biological sciences. Psychology Humans Molecular and cellular biology Monocytes - immunology Monocytes - pathology |
title | Surface Molecule Loss and Bleb Formation by Human Germinal Center B Cells Undergoing Apoptosis: Role of Apoptotic Blebs in Monocyte Chemotaxis |
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