Activation‐induced CD137 is a fast assay for identification and multi‐parameter flow cytometric analysis of alloreactive T cells
Summary Detection and isolation of viable alloreactive T cells at the single‐cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)‐family, CD137 (4‐1BB) was investigated for its potential...
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| Veröffentlicht in: | Clinical and experimental immunology 2013-10, Vol.174 (1), p.179-191 |
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| creator | Litjens, N. H. R. Wit, E. A. Baan, C. C. Betjes, M. G. H. |
| description | Summary
Detection and isolation of viable alloreactive T cells at the single‐cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)‐family, CD137 (4‐1BB) was investigated for its potential to identify the total pool of circulating alloreactive T cells. Optimal conditions for sensitive and specific detection of allogeneic‐induced CD137 expression on circulating T cells were established. Thereafter, CD137+ alloreactive T cells were phenotypically and functionally characterized by multi‐parameter flow cytometry. Alloantigen‐induced CD137 expression identified both alloreactive CD8+ T cells (mean ± standard error of the mean: 0·21 ± 0·07%) and alloreactive CD4+ T cells (0·21 ± 0·05%). CD137+ alloreactive T cells were detected in different T cell subsets, including naive T cells, but were found preferentially in CD28+ T cells and not in the terminally differentiated T cell subset. Upon allogeneic (re‐)stimulation, the cytokine‐producing as well as proliferative capacity of T cells resided mainly within the CD137‐expressing fraction. About 10% of the CD137+ alloreactive T cells produced any combination of interferon (IFN)‐γ, interleukin (IL)‐2 and TNF‐α. Polyfunctional alloreactive T cells, defined by multiple cytokine expression, were observed infrequently. In conclusion, activation‐induced CD137 expression is a fast assay allowing for detection and functional analysis of the total alloreactive T cell compartment at the single‐cell level by multi‐parameter flow cytometry. |
| doi_str_mv | 10.1111/cei.12152 |
| format | Article |
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Detection and isolation of viable alloreactive T cells at the single‐cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)‐family, CD137 (4‐1BB) was investigated for its potential to identify the total pool of circulating alloreactive T cells. Optimal conditions for sensitive and specific detection of allogeneic‐induced CD137 expression on circulating T cells were established. Thereafter, CD137+ alloreactive T cells were phenotypically and functionally characterized by multi‐parameter flow cytometry. Alloantigen‐induced CD137 expression identified both alloreactive CD8+ T cells (mean ± standard error of the mean: 0·21 ± 0·07%) and alloreactive CD4+ T cells (0·21 ± 0·05%). CD137+ alloreactive T cells were detected in different T cell subsets, including naive T cells, but were found preferentially in CD28+ T cells and not in the terminally differentiated T cell subset. Upon allogeneic (re‐)stimulation, the cytokine‐producing as well as proliferative capacity of T cells resided mainly within the CD137‐expressing fraction. About 10% of the CD137+ alloreactive T cells produced any combination of interferon (IFN)‐γ, interleukin (IL)‐2 and TNF‐α. Polyfunctional alloreactive T cells, defined by multiple cytokine expression, were observed infrequently. In conclusion, activation‐induced CD137 expression is a fast assay allowing for detection and functional analysis of the total alloreactive T cell compartment at the single‐cell level by multi‐parameter flow cytometry.</description><identifier>ISSN: 0009-9104</identifier><identifier>EISSN: 1365-2249</identifier><identifier>DOI: 10.1111/cei.12152</identifier><identifier>PMID: 23750604</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>alloreactivity ; Antigen-Presenting Cells - cytology ; Antigen-Presenting Cells - immunology ; Antigen-Presenting Cells - metabolism ; CD137 ; CD137 antigen ; CD4+ T cells ; CD4-Positive T-Lymphocytes - cytology ; CD4-Positive T-Lymphocytes - immunology ; CD4-Positive T-Lymphocytes - metabolism ; CD8+ T cells ; CD8-Positive T-Lymphocytes - cytology ; CD8-Positive T-Lymphocytes - immunology ; CD8-Positive T-Lymphocytes - metabolism ; Cell Proliferation ; Flow Cytometry - methods ; Flow Cytometry - standards ; Humans ; Immunophenotyping - methods ; Immunophenotyping - standards ; Isoantigens - physiology ; Lymphocyte Activation - immunology ; Lymphocyte Culture Test, Mixed - methods ; Lymphocyte Culture Test, Mixed - standards ; Lymphocyte Depletion ; multi‐parameter analysis ; Original ; Sensitivity and Specificity ; Stem Cells - cytology ; Stem Cells - immunology ; Stem Cells - metabolism ; T-Lymphocyte Subsets - cytology ; T-Lymphocyte Subsets - immunology ; T-Lymphocyte Subsets - metabolism ; Tumor Necrosis Factor Receptor Superfamily, Member 9 - biosynthesis ; Tumor Necrosis Factor Receptor Superfamily, Member 9 - genetics</subject><ispartof>Clinical and experimental immunology, 2013-10, Vol.174 (1), p.179-191</ispartof><rights>2013 British Society for Immunology</rights><rights>2013 British Society for Immunology.</rights><rights>Copyright © 2013 British Society for Immunology</rights><rights>Copyright © 2013 British Society for Immunology 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4762-a4e381eb96c05f0fc379b532fc091c5ec36d9f7aabf6544e23df5e57bd7ac71a3</citedby><cites>FETCH-LOGICAL-c4762-a4e381eb96c05f0fc379b532fc091c5ec36d9f7aabf6544e23df5e57bd7ac71a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3784225/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3784225/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23750604$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Litjens, N. H. R.</creatorcontrib><creatorcontrib>Wit, E. A.</creatorcontrib><creatorcontrib>Baan, C. C.</creatorcontrib><creatorcontrib>Betjes, M. G. H.</creatorcontrib><title>Activation‐induced CD137 is a fast assay for identification and multi‐parameter flow cytometric analysis of alloreactive T cells</title><title>Clinical and experimental immunology</title><addtitle>Clin Exp Immunol</addtitle><description>Summary
Detection and isolation of viable alloreactive T cells at the single‐cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)‐family, CD137 (4‐1BB) was investigated for its potential to identify the total pool of circulating alloreactive T cells. Optimal conditions for sensitive and specific detection of allogeneic‐induced CD137 expression on circulating T cells were established. Thereafter, CD137+ alloreactive T cells were phenotypically and functionally characterized by multi‐parameter flow cytometry. Alloantigen‐induced CD137 expression identified both alloreactive CD8+ T cells (mean ± standard error of the mean: 0·21 ± 0·07%) and alloreactive CD4+ T cells (0·21 ± 0·05%). CD137+ alloreactive T cells were detected in different T cell subsets, including naive T cells, but were found preferentially in CD28+ T cells and not in the terminally differentiated T cell subset. Upon allogeneic (re‐)stimulation, the cytokine‐producing as well as proliferative capacity of T cells resided mainly within the CD137‐expressing fraction. About 10% of the CD137+ alloreactive T cells produced any combination of interferon (IFN)‐γ, interleukin (IL)‐2 and TNF‐α. Polyfunctional alloreactive T cells, defined by multiple cytokine expression, were observed infrequently. In conclusion, activation‐induced CD137 expression is a fast assay allowing for detection and functional analysis of the total alloreactive T cell compartment at the single‐cell level by multi‐parameter flow cytometry.</description><subject>alloreactivity</subject><subject>Antigen-Presenting Cells - cytology</subject><subject>Antigen-Presenting Cells - immunology</subject><subject>Antigen-Presenting Cells - metabolism</subject><subject>CD137</subject><subject>CD137 antigen</subject><subject>CD4+ T cells</subject><subject>CD4-Positive T-Lymphocytes - cytology</subject><subject>CD4-Positive T-Lymphocytes - immunology</subject><subject>CD4-Positive T-Lymphocytes - metabolism</subject><subject>CD8+ T cells</subject><subject>CD8-Positive T-Lymphocytes - cytology</subject><subject>CD8-Positive T-Lymphocytes - immunology</subject><subject>CD8-Positive T-Lymphocytes - metabolism</subject><subject>Cell Proliferation</subject><subject>Flow Cytometry - methods</subject><subject>Flow Cytometry - standards</subject><subject>Humans</subject><subject>Immunophenotyping - methods</subject><subject>Immunophenotyping - standards</subject><subject>Isoantigens - physiology</subject><subject>Lymphocyte Activation - immunology</subject><subject>Lymphocyte Culture Test, Mixed - methods</subject><subject>Lymphocyte Culture Test, Mixed - standards</subject><subject>Lymphocyte Depletion</subject><subject>multi‐parameter analysis</subject><subject>Original</subject><subject>Sensitivity and Specificity</subject><subject>Stem Cells - cytology</subject><subject>Stem Cells - immunology</subject><subject>Stem Cells - metabolism</subject><subject>T-Lymphocyte Subsets - cytology</subject><subject>T-Lymphocyte Subsets - immunology</subject><subject>T-Lymphocyte Subsets - metabolism</subject><subject>Tumor Necrosis Factor Receptor Superfamily, Member 9 - biosynthesis</subject><subject>Tumor Necrosis Factor Receptor Superfamily, Member 9 - genetics</subject><issn>0009-9104</issn><issn>1365-2249</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNks9qFTEUxoMo9lpd-AIScFMX0-Z_7myEcq1aKLip63Amk2hKZnJNZlpm58IH8Bl9EnN7a1FBMJtwyO98J1_yIfSckmNa14l14ZgyKtkDtKJcyYYx0T5EK0JI27SUiAP0pJSrWiql2GN0wLiWRBGxQt9O7RSuYQpp_PH1exj72boeb95QrnEoGLCHMmEoBRbsU8ahd-MUfLC3LRjGHg9znEJt3kKGwU0uYx_TDbbLlGqZg60UxKVUueQxxJiyg91Uhy-xdTGWp-iRh1jcs7v9EH18e3a5ed9cfHh3vjm9aKzQijUgHF9T17XKEumJt1y3neTMW9JSK53lqm-9Bui8kkI4xnsvndRdr8FqCvwQvd7rbuducL2tVjJEs81hgLyYBMH8eTKGz-ZTujZcrwVjsgoc3Qnk9GV2ZTJDKDsLMLo0F0OFXEvBiWb_gXLKWk2krujLv9CrNOf6ZpWSrK1WOOOVerWnbE6lZOfv702J2cXA1BiY2xhU9sXvRu_JX_9egZM9cBOiW_6tZDZn53vJn6cxwBs</recordid><startdate>201310</startdate><enddate>201310</enddate><creator>Litjens, N. H. R.</creator><creator>Wit, E. A.</creator><creator>Baan, C. C.</creator><creator>Betjes, M. G. H.</creator><general>Oxford University Press</general><general>Blackwell Science Inc</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>7T5</scope><scope>7U9</scope><scope>H94</scope><scope>M7N</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>201310</creationdate><title>Activation‐induced CD137 is a fast assay for identification and multi‐parameter flow cytometric analysis of alloreactive T cells</title><author>Litjens, N. H. R. ; Wit, E. A. ; Baan, C. C. ; Betjes, M. G. H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4762-a4e381eb96c05f0fc379b532fc091c5ec36d9f7aabf6544e23df5e57bd7ac71a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>alloreactivity</topic><topic>Antigen-Presenting Cells - cytology</topic><topic>Antigen-Presenting Cells - immunology</topic><topic>Antigen-Presenting Cells - metabolism</topic><topic>CD137</topic><topic>CD137 antigen</topic><topic>CD4+ T cells</topic><topic>CD4-Positive T-Lymphocytes - cytology</topic><topic>CD4-Positive T-Lymphocytes - immunology</topic><topic>CD4-Positive T-Lymphocytes - metabolism</topic><topic>CD8+ T cells</topic><topic>CD8-Positive T-Lymphocytes - cytology</topic><topic>CD8-Positive T-Lymphocytes - immunology</topic><topic>CD8-Positive T-Lymphocytes - metabolism</topic><topic>Cell Proliferation</topic><topic>Flow Cytometry - methods</topic><topic>Flow Cytometry - standards</topic><topic>Humans</topic><topic>Immunophenotyping - methods</topic><topic>Immunophenotyping - standards</topic><topic>Isoantigens - physiology</topic><topic>Lymphocyte Activation - immunology</topic><topic>Lymphocyte Culture Test, Mixed - methods</topic><topic>Lymphocyte Culture Test, Mixed - standards</topic><topic>Lymphocyte Depletion</topic><topic>multi‐parameter analysis</topic><topic>Original</topic><topic>Sensitivity and Specificity</topic><topic>Stem Cells - cytology</topic><topic>Stem Cells - immunology</topic><topic>Stem Cells - metabolism</topic><topic>T-Lymphocyte Subsets - cytology</topic><topic>T-Lymphocyte Subsets - immunology</topic><topic>T-Lymphocyte Subsets - metabolism</topic><topic>Tumor Necrosis Factor Receptor Superfamily, Member 9 - biosynthesis</topic><topic>Tumor Necrosis Factor Receptor Superfamily, Member 9 - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Litjens, N. H. R.</creatorcontrib><creatorcontrib>Wit, E. A.</creatorcontrib><creatorcontrib>Baan, C. C.</creatorcontrib><creatorcontrib>Betjes, M. G. H.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Clinical and experimental immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Litjens, N. H. R.</au><au>Wit, E. A.</au><au>Baan, C. C.</au><au>Betjes, M. G. H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Activation‐induced CD137 is a fast assay for identification and multi‐parameter flow cytometric analysis of alloreactive T cells</atitle><jtitle>Clinical and experimental immunology</jtitle><addtitle>Clin Exp Immunol</addtitle><date>2013-10</date><risdate>2013</risdate><volume>174</volume><issue>1</issue><spage>179</spage><epage>191</epage><pages>179-191</pages><issn>0009-9104</issn><eissn>1365-2249</eissn><abstract>Summary
Detection and isolation of viable alloreactive T cells at the single‐cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)‐family, CD137 (4‐1BB) was investigated for its potential to identify the total pool of circulating alloreactive T cells. Optimal conditions for sensitive and specific detection of allogeneic‐induced CD137 expression on circulating T cells were established. Thereafter, CD137+ alloreactive T cells were phenotypically and functionally characterized by multi‐parameter flow cytometry. Alloantigen‐induced CD137 expression identified both alloreactive CD8+ T cells (mean ± standard error of the mean: 0·21 ± 0·07%) and alloreactive CD4+ T cells (0·21 ± 0·05%). CD137+ alloreactive T cells were detected in different T cell subsets, including naive T cells, but were found preferentially in CD28+ T cells and not in the terminally differentiated T cell subset. Upon allogeneic (re‐)stimulation, the cytokine‐producing as well as proliferative capacity of T cells resided mainly within the CD137‐expressing fraction. About 10% of the CD137+ alloreactive T cells produced any combination of interferon (IFN)‐γ, interleukin (IL)‐2 and TNF‐α. Polyfunctional alloreactive T cells, defined by multiple cytokine expression, were observed infrequently. In conclusion, activation‐induced CD137 expression is a fast assay allowing for detection and functional analysis of the total alloreactive T cell compartment at the single‐cell level by multi‐parameter flow cytometry.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>23750604</pmid><doi>10.1111/cei.12152</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | alloreactivity Antigen-Presenting Cells - cytology Antigen-Presenting Cells - immunology Antigen-Presenting Cells - metabolism CD137 CD137 antigen CD4+ T cells CD4-Positive T-Lymphocytes - cytology CD4-Positive T-Lymphocytes - immunology CD4-Positive T-Lymphocytes - metabolism CD8+ T cells CD8-Positive T-Lymphocytes - cytology CD8-Positive T-Lymphocytes - immunology CD8-Positive T-Lymphocytes - metabolism Cell Proliferation Flow Cytometry - methods Flow Cytometry - standards Humans Immunophenotyping - methods Immunophenotyping - standards Isoantigens - physiology Lymphocyte Activation - immunology Lymphocyte Culture Test, Mixed - methods Lymphocyte Culture Test, Mixed - standards Lymphocyte Depletion multi‐parameter analysis Original Sensitivity and Specificity Stem Cells - cytology Stem Cells - immunology Stem Cells - metabolism T-Lymphocyte Subsets - cytology T-Lymphocyte Subsets - immunology T-Lymphocyte Subsets - metabolism Tumor Necrosis Factor Receptor Superfamily, Member 9 - biosynthesis Tumor Necrosis Factor Receptor Superfamily, Member 9 - genetics |
| title | Activation‐induced CD137 is a fast assay for identification and multi‐parameter flow cytometric analysis of alloreactive T cells |
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