Measurement of T Cell Activation After 16‐hr In Vitro Stimulation with Concanavalin A
A flow cytometry assay that can be used to directly determine the proportion of activated T lymphocytes in human whole blood samples after stimulation with concanavalin A is presented here. Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythr...
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| Veröffentlicht in: | Current Protocols in Cytometry 2010-01, Vol.51 (1), p.6.28.1-6.28.10 |
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| container_title | Current Protocols in Cytometry |
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| creator | Arneth, Borros M. |
| description | A flow cytometry assay that can be used to directly determine the proportion of activated T lymphocytes in human whole blood samples after stimulation with concanavalin A is presented here. Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre‐analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments. Curr. Protoc. Cytom. 51:6.28.1‐6.28.10. © 2010 by John Wiley & Sons, Inc. |
| doi_str_mv | 10.1002/0471142956.cy0628s51 |
| format | Article |
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Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre‐analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments. Curr. Protoc. 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Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre‐analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments. Curr. Protoc. Cytom. 51:6.28.1‐6.28.10. © 2010 by John Wiley & Sons, Inc.</description><subject>activated T cells</subject><subject>activation marker</subject><subject>Antibodies, Monoclonal - pharmacology</subject><subject>CD4 T cells</subject><subject>CD4 T helper cells</subject><subject>CD4-Positive T-Lymphocytes - drug effects</subject><subject>CD4-Positive T-Lymphocytes - immunology</subject><subject>CD69</subject><subject>CD69 activation marker</subject><subject>CD8 cytotoxic T cells</subject><subject>CD8-Positive T-Lymphocytes - drug effects</subject><subject>CD8-Positive T-Lymphocytes - immunology</subject><subject>Cell Extracts</subject><subject>Concanavalin A - immunology</subject><subject>Concanavalin A - pharmacology</subject><subject>Erythrocytes - drug effects</subject><subject>Flow Cytometry - methods</subject><subject>Humans</subject><subject>Immunophenotyping</subject><subject>Lymphocyte Activation - drug effects</subject><subject>lymphocyte proliferation assay</subject><subject>Lymphocyte Subsets - drug effects</subject><subject>lymphocytes</subject><subject>T lymphocytes</subject><subject>Time Factors</subject><issn>1934-9297</issn><issn>1934-9300</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkMtKw0AYhQdRbKl9A5F5gdR_Lplk3JXgpVBRsCquwmQyQwdyKZO0pTsfwWf0SUyJrVtX5yzOdxYfQpcEJgSAXgOPCOFUhmKidyBo3ITkBA2JZDyQDOD00KmMBmjcNC4D4JRJEOwcDSiAkCEVQ_T-aFSz9qY0VYtrixc4MUWBp7p1G9W6usJT2xqPifj-_Fp6PKvwm2t9jV9aV66LfrJ17RIndaVVpTaqcB10gc6sKhoz_s0Rer27XSQPwfzpfpZM54GmEY0DqrXNszADJWVuQsXBRlmecZBacy7imFopcmtizpRQHRFT0fVYWZNbrXM2Qrz_1b5uGm9suvKuVH6XEkj3qtI_VelRVYdd9dhqnZUmP0IHMd3gph9sXWF2_zpNk-fkY9_ZD-N3d2Q</recordid><startdate>201001</startdate><enddate>201001</enddate><creator>Arneth, Borros M.</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>201001</creationdate><title>Measurement of T Cell Activation After 16‐hr In Vitro Stimulation with Concanavalin A</title><author>Arneth, Borros M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2728-2ccfdb5b0a99de5a40f7bdb409cc446882f96dfe843a6a7288268438afedfccd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>activated T cells</topic><topic>activation marker</topic><topic>Antibodies, Monoclonal - pharmacology</topic><topic>CD4 T cells</topic><topic>CD4 T helper cells</topic><topic>CD4-Positive T-Lymphocytes - drug effects</topic><topic>CD4-Positive T-Lymphocytes - immunology</topic><topic>CD69</topic><topic>CD69 activation marker</topic><topic>CD8 cytotoxic T cells</topic><topic>CD8-Positive T-Lymphocytes - drug effects</topic><topic>CD8-Positive T-Lymphocytes - immunology</topic><topic>Cell Extracts</topic><topic>Concanavalin A - immunology</topic><topic>Concanavalin A - pharmacology</topic><topic>Erythrocytes - drug effects</topic><topic>Flow Cytometry - methods</topic><topic>Humans</topic><topic>Immunophenotyping</topic><topic>Lymphocyte Activation - drug effects</topic><topic>lymphocyte proliferation assay</topic><topic>Lymphocyte Subsets - drug effects</topic><topic>lymphocytes</topic><topic>T lymphocytes</topic><topic>Time Factors</topic><toplevel>online_resources</toplevel><creatorcontrib>Arneth, Borros M.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>Current Protocols in Cytometry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Arneth, Borros M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Measurement of T Cell Activation After 16‐hr In Vitro Stimulation with Concanavalin A</atitle><jtitle>Current Protocols in Cytometry</jtitle><addtitle>Curr Protoc Cytom</addtitle><date>2010-01</date><risdate>2010</risdate><volume>51</volume><issue>1</issue><spage>6.28.1</spage><epage>6.28.10</epage><pages>6.28.1-6.28.10</pages><issn>1934-9297</issn><eissn>1934-9300</eissn><abstract>A flow cytometry assay that can be used to directly determine the proportion of activated T lymphocytes in human whole blood samples after stimulation with concanavalin A is presented here. Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre‐analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments. Curr. Protoc. Cytom. 51:6.28.1‐6.28.10. © 2010 by John Wiley & Sons, Inc.</abstract><cop>United States</cop><pmid>20069526</pmid><doi>10.1002/0471142956.cy0628s51</doi><tpages>10</tpages></addata></record> |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | activated T cells activation marker Antibodies, Monoclonal - pharmacology CD4 T cells CD4 T helper cells CD4-Positive T-Lymphocytes - drug effects CD4-Positive T-Lymphocytes - immunology CD69 CD69 activation marker CD8 cytotoxic T cells CD8-Positive T-Lymphocytes - drug effects CD8-Positive T-Lymphocytes - immunology Cell Extracts Concanavalin A - immunology Concanavalin A - pharmacology Erythrocytes - drug effects Flow Cytometry - methods Humans Immunophenotyping Lymphocyte Activation - drug effects lymphocyte proliferation assay Lymphocyte Subsets - drug effects lymphocytes T lymphocytes Time Factors |
| title | Measurement of T Cell Activation After 16‐hr In Vitro Stimulation with Concanavalin A |
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