Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity
A major goal in immunodiagnostics has been the development of assay systems that can measure CD8 + T cell immunity in humans, directly ex vivo, at high resolution, and with high throughput. We established granzyme B (grB) enzyme-linked immunospot assay (ELISPOT) in conjunction with image analysis to...
Gespeichert in:
| Veröffentlicht in: | Journal of immunological methods 2000-06, Vol.240 (1), p.143-155 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 155 |
|---|---|
| container_issue | 1 |
| container_start_page | 143 |
| container_title | Journal of immunological methods |
| container_volume | 240 |
| creator | Rininsland, Frauke H Helms, Thomas Asaad, Robert J Boehm, Bernhard O Tary-Lehmann, Magdalena |
| description | A major goal in immunodiagnostics has been the development of assay systems that can measure CD8
+ T cell immunity in humans, directly ex vivo, at high resolution, and with high throughput. We established granzyme B (grB) enzyme-linked immunospot assay (ELISPOT) in conjunction with image analysis to this end. Using grB transfected and untransfected Chinese hamster ovary (CHO) cells and T cell lines, we show that the antibody pair utilized was grB-specific and that only activated T cells secrete grB. GrB release began within 4 h after antigen stimulation and stopped within 40 h. Side-by-side comparison showed grB ELISPOT assays to have a higher resolution than classic chromium-release assays in terms of signal-to-noise ratio. The linearity of the relation of the number of CD8
+ effector T cells plated to grB spots detected suggests that grB ELISPOT assays measure the frequencies of grB-secreting cells directly. Reactivity to HIV peptides was seen in grB ELISPOT assays of freshly isolated PBMC from HIV patients, consistent with the detection of peptide-specific memory cells. The higher resolution and lower labor and material investment should make grB ELISPOT assays an attractive alternative to chromium-release assays in monitoring the clonal sizes of specific CD8 memory cells in vivo. |
| doi_str_mv | 10.1016/S0022-1759(00)00191-5 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_17598719</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0022175900001915</els_id><sourcerecordid>17598719</sourcerecordid><originalsourceid>FETCH-LOGICAL-c420t-9c6881884e10f9c11babc7f30293a827ca5cda6134828e101b283269e1f9161d3</originalsourceid><addsrcrecordid>eNqF0FtL5DAUwPEgLjpePoISEBZ96O45adMmT-JtXWFghBmfQyY9hci01aQddvbT27kgvvmUl99JTv6MnSH8QsD89xRAiAQLqS8BrgBQYyL32AhVIZJCg9xno09yyI5ifIVBQQ4H7BBBySwHPWL3j8E2_1c18Vv-MH6aPk9m3MZoV7xqA6d_fOmXLa_Jxj5QTU0XeVvxGXe0WHBf133ju9UJ-1HZRaTT3XnMXv48zO7-JuPJ49PdzThxmYAu0S5XCpXKCKHSDnFu566oUhA6tUoUzkpX2hzTTAk1GJwLlYpcE1YacyzTY_Zze-9baN97ip2pfVxvYhtq-2jWX1UF6gHKLXShjTFQZd6Cr21YGQSzzmc2-TYDBsBs8hk5zJ3vHujnNZVfpra9BnCxAzY6u6iGeM7HT6eyQsp8UNdbRUOMpadgovPUOCp9INeZsvXfLPIBojCJLw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17598719</pqid></control><display><type>article</type><title>Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals Complete</source><creator>Rininsland, Frauke H ; Helms, Thomas ; Asaad, Robert J ; Boehm, Bernhard O ; Tary-Lehmann, Magdalena</creator><creatorcontrib>Rininsland, Frauke H ; Helms, Thomas ; Asaad, Robert J ; Boehm, Bernhard O ; Tary-Lehmann, Magdalena</creatorcontrib><description>A major goal in immunodiagnostics has been the development of assay systems that can measure CD8
+ T cell immunity in humans, directly ex vivo, at high resolution, and with high throughput. We established granzyme B (grB) enzyme-linked immunospot assay (ELISPOT) in conjunction with image analysis to this end. Using grB transfected and untransfected Chinese hamster ovary (CHO) cells and T cell lines, we show that the antibody pair utilized was grB-specific and that only activated T cells secrete grB. GrB release began within 4 h after antigen stimulation and stopped within 40 h. Side-by-side comparison showed grB ELISPOT assays to have a higher resolution than classic chromium-release assays in terms of signal-to-noise ratio. The linearity of the relation of the number of CD8
+ effector T cells plated to grB spots detected suggests that grB ELISPOT assays measure the frequencies of grB-secreting cells directly. Reactivity to HIV peptides was seen in grB ELISPOT assays of freshly isolated PBMC from HIV patients, consistent with the detection of peptide-specific memory cells. The higher resolution and lower labor and material investment should make grB ELISPOT assays an attractive alternative to chromium-release assays in monitoring the clonal sizes of specific CD8 memory cells in vivo.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/S0022-1759(00)00191-5</identifier><identifier>PMID: 10854609</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; CD8 antigen ; Cell Line ; CHO Cells ; Chromium ; Cloning, Molecular ; Cricetinae ; Cytotoxic T cells ; Cytotoxicity Tests, Immunologic - methods ; ELISPOT ; Enzyme-Linked Immunosorbent Assay - methods ; Flow Cytometry ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Granzyme B ; Granzymes ; HIV Antigens - immunology ; Humans ; Immunologic Memory ; Lymphocyte Activation ; Memory ; Molecular immunology ; Molecular Sequence Data ; Peptide-specificity ; Serine Endopeptidases - genetics ; Serine Endopeptidases - metabolism ; T-Lymphocytes, Cytotoxic - cytology ; T-Lymphocytes, Cytotoxic - immunology ; Techniques</subject><ispartof>Journal of immunological methods, 2000-06, Vol.240 (1), p.143-155</ispartof><rights>2000 Elsevier Science B.V.</rights><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c420t-9c6881884e10f9c11babc7f30293a827ca5cda6134828e101b283269e1f9161d3</citedby><cites>FETCH-LOGICAL-c420t-9c6881884e10f9c11babc7f30293a827ca5cda6134828e101b283269e1f9161d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0022175900001915$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=847556$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10854609$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rininsland, Frauke H</creatorcontrib><creatorcontrib>Helms, Thomas</creatorcontrib><creatorcontrib>Asaad, Robert J</creatorcontrib><creatorcontrib>Boehm, Bernhard O</creatorcontrib><creatorcontrib>Tary-Lehmann, Magdalena</creatorcontrib><title>Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>A major goal in immunodiagnostics has been the development of assay systems that can measure CD8
+ T cell immunity in humans, directly ex vivo, at high resolution, and with high throughput. We established granzyme B (grB) enzyme-linked immunospot assay (ELISPOT) in conjunction with image analysis to this end. Using grB transfected and untransfected Chinese hamster ovary (CHO) cells and T cell lines, we show that the antibody pair utilized was grB-specific and that only activated T cells secrete grB. GrB release began within 4 h after antigen stimulation and stopped within 40 h. Side-by-side comparison showed grB ELISPOT assays to have a higher resolution than classic chromium-release assays in terms of signal-to-noise ratio. The linearity of the relation of the number of CD8
+ effector T cells plated to grB spots detected suggests that grB ELISPOT assays measure the frequencies of grB-secreting cells directly. Reactivity to HIV peptides was seen in grB ELISPOT assays of freshly isolated PBMC from HIV patients, consistent with the detection of peptide-specific memory cells. The higher resolution and lower labor and material investment should make grB ELISPOT assays an attractive alternative to chromium-release assays in monitoring the clonal sizes of specific CD8 memory cells in vivo.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>CD8 antigen</subject><subject>Cell Line</subject><subject>CHO Cells</subject><subject>Chromium</subject><subject>Cloning, Molecular</subject><subject>Cricetinae</subject><subject>Cytotoxic T cells</subject><subject>Cytotoxicity Tests, Immunologic - methods</subject><subject>ELISPOT</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Flow Cytometry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Granzyme B</subject><subject>Granzymes</subject><subject>HIV Antigens - immunology</subject><subject>Humans</subject><subject>Immunologic Memory</subject><subject>Lymphocyte Activation</subject><subject>Memory</subject><subject>Molecular immunology</subject><subject>Molecular Sequence Data</subject><subject>Peptide-specificity</subject><subject>Serine Endopeptidases - genetics</subject><subject>Serine Endopeptidases - metabolism</subject><subject>T-Lymphocytes, Cytotoxic - cytology</subject><subject>T-Lymphocytes, Cytotoxic - immunology</subject><subject>Techniques</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0FtL5DAUwPEgLjpePoISEBZ96O45adMmT-JtXWFghBmfQyY9hci01aQddvbT27kgvvmUl99JTv6MnSH8QsD89xRAiAQLqS8BrgBQYyL32AhVIZJCg9xno09yyI5ifIVBQQ4H7BBBySwHPWL3j8E2_1c18Vv-MH6aPk9m3MZoV7xqA6d_fOmXLa_Jxj5QTU0XeVvxGXe0WHBf133ju9UJ-1HZRaTT3XnMXv48zO7-JuPJ49PdzThxmYAu0S5XCpXKCKHSDnFu566oUhA6tUoUzkpX2hzTTAk1GJwLlYpcE1YacyzTY_Zze-9baN97ip2pfVxvYhtq-2jWX1UF6gHKLXShjTFQZd6Cr21YGQSzzmc2-TYDBsBs8hk5zJ3vHujnNZVfpra9BnCxAzY6u6iGeM7HT6eyQsp8UNdbRUOMpadgovPUOCp9INeZsvXfLPIBojCJLw</recordid><startdate>20000623</startdate><enddate>20000623</enddate><creator>Rininsland, Frauke H</creator><creator>Helms, Thomas</creator><creator>Asaad, Robert J</creator><creator>Boehm, Bernhard O</creator><creator>Tary-Lehmann, Magdalena</creator><general>Elsevier B.V</general><general>Elsevier</general><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>7QO</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope></search><sort><creationdate>20000623</creationdate><title>Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity</title><author>Rininsland, Frauke H ; Helms, Thomas ; Asaad, Robert J ; Boehm, Bernhard O ; Tary-Lehmann, Magdalena</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c420t-9c6881884e10f9c11babc7f30293a827ca5cda6134828e101b283269e1f9161d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>CD8 antigen</topic><topic>Cell Line</topic><topic>CHO Cells</topic><topic>Chromium</topic><topic>Cloning, Molecular</topic><topic>Cricetinae</topic><topic>Cytotoxic T cells</topic><topic>Cytotoxicity Tests, Immunologic - methods</topic><topic>ELISPOT</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Flow Cytometry</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Granzyme B</topic><topic>Granzymes</topic><topic>HIV Antigens - immunology</topic><topic>Humans</topic><topic>Immunologic Memory</topic><topic>Lymphocyte Activation</topic><topic>Memory</topic><topic>Molecular immunology</topic><topic>Molecular Sequence Data</topic><topic>Peptide-specificity</topic><topic>Serine Endopeptidases - genetics</topic><topic>Serine Endopeptidases - metabolism</topic><topic>T-Lymphocytes, Cytotoxic - cytology</topic><topic>T-Lymphocytes, Cytotoxic - immunology</topic><topic>Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rininsland, Frauke H</creatorcontrib><creatorcontrib>Helms, Thomas</creatorcontrib><creatorcontrib>Asaad, Robert J</creatorcontrib><creatorcontrib>Boehm, Bernhard O</creatorcontrib><creatorcontrib>Tary-Lehmann, Magdalena</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>Biotechnology Research Abstracts</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rininsland, Frauke H</au><au>Helms, Thomas</au><au>Asaad, Robert J</au><au>Boehm, Bernhard O</au><au>Tary-Lehmann, Magdalena</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>2000-06-23</date><risdate>2000</risdate><volume>240</volume><issue>1</issue><spage>143</spage><epage>155</epage><pages>143-155</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>A major goal in immunodiagnostics has been the development of assay systems that can measure CD8
+ T cell immunity in humans, directly ex vivo, at high resolution, and with high throughput. We established granzyme B (grB) enzyme-linked immunospot assay (ELISPOT) in conjunction with image analysis to this end. Using grB transfected and untransfected Chinese hamster ovary (CHO) cells and T cell lines, we show that the antibody pair utilized was grB-specific and that only activated T cells secrete grB. GrB release began within 4 h after antigen stimulation and stopped within 40 h. Side-by-side comparison showed grB ELISPOT assays to have a higher resolution than classic chromium-release assays in terms of signal-to-noise ratio. The linearity of the relation of the number of CD8
+ effector T cells plated to grB spots detected suggests that grB ELISPOT assays measure the frequencies of grB-secreting cells directly. Reactivity to HIV peptides was seen in grB ELISPOT assays of freshly isolated PBMC from HIV patients, consistent with the detection of peptide-specific memory cells. The higher resolution and lower labor and material investment should make grB ELISPOT assays an attractive alternative to chromium-release assays in monitoring the clonal sizes of specific CD8 memory cells in vivo.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>10854609</pmid><doi>10.1016/S0022-1759(00)00191-5</doi><tpages>13</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-1759 |
| ispartof | Journal of immunological methods, 2000-06, Vol.240 (1), p.143-155 |
| issn | 0022-1759 1872-7905 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17598719 |
| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Animals Base Sequence Biological and medical sciences CD8 antigen Cell Line CHO Cells Chromium Cloning, Molecular Cricetinae Cytotoxic T cells Cytotoxicity Tests, Immunologic - methods ELISPOT Enzyme-Linked Immunosorbent Assay - methods Flow Cytometry Fundamental and applied biological sciences. Psychology Fundamental immunology Granzyme B Granzymes HIV Antigens - immunology Humans Immunologic Memory Lymphocyte Activation Memory Molecular immunology Molecular Sequence Data Peptide-specificity Serine Endopeptidases - genetics Serine Endopeptidases - metabolism T-Lymphocytes, Cytotoxic - cytology T-Lymphocytes, Cytotoxic - immunology Techniques |
| title | Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-21T21%3A02%3A47IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Granzyme%20B%20ELISPOT%20assay%20for%20ex%20vivo%20measurements%20of%20T%20cell%20immunity&rft.jtitle=Journal%20of%20immunological%20methods&rft.au=Rininsland,%20Frauke%20H&rft.date=2000-06-23&rft.volume=240&rft.issue=1&rft.spage=143&rft.epage=155&rft.pages=143-155&rft.issn=0022-1759&rft.eissn=1872-7905&rft.coden=JIMMBG&rft_id=info:doi/10.1016/S0022-1759(00)00191-5&rft_dat=%3Cproquest_cross%3E17598719%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17598719&rft_id=info:pmid/10854609&rft_els_id=S0022175900001915&rfr_iscdi=true |