Multicenter study on in vitro characterization of dendritic cells
Background There is growing interest in the use of in vitro -expanded dendritic cells (DC) in cancer immunotherapy as cellular-based vaccines. However, the methods used for in vitro preparation vary widely between institutions. Therefore, a strong need exists for standardization, characterization an...
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| Veröffentlicht in: | Cytotherapy (Oxford, England) England), 2008-01, Vol.10 (1), p.21-29 |
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| creator | Eichler, H Nguyen, X.D Roelen, D Celluzzi, C.M McKenna, D Pamphilon, D Blair, A Read, E.J Takahashi, T.A Szczepiorkowski, Z.M |
| description | Background There is growing interest in the use of in vitro -expanded dendritic cells (DC) in cancer immunotherapy as cellular-based vaccines. However, the methods used for in vitro preparation vary widely between institutions. Therefore, a strong need exists for standardization, characterization and quality control (QC) of such vaccines. A first prospective multicenter pilot study was performed to investigate basic QC parameters of frozen/thawed DC. The study design was focused on comparison of test results for cell counts, immunophenotyping and cell viability. Methods CD14+ monocytes were isolated from three healthy volunteers. The cells were expanded in vitro , matured and cryopreserved using a standardized protocol in one laboratory. The aliquots of cryopreserved DC and a panel of reagents were shipped to eight laboratories worldwide. The objective was to compare the results of non-functional QC assays between sites by testing identical DC vaccines and using a pre-defined test protocol. Results Measurements of nucleated cell (NC) content of thawed DC vaccines with different types of hematology analyzers (HA) gave similar results for the majority of sites. Immunophenotyping using identical clones of monoclonal antibodies for the detection of surface antigens (i.e. CD1a, CD14, CD16, CD83, CD86 and HLA-DR) provided mostly comparable results between laboratories with an acceptable level of variation. In contrast, highly different results between study sites were generated for measuring the viability of thawed DC by flow cytometry using 7-amino-actinomycin D (7-AAD) dye exclusion. Discussion In characterizing frozen/thawed DC vaccines, NC counts generated by HA yielded similar results between different laboratories. Furthermore, immunophenotyping of DC vaccines can be standardized between centers, i.e. by using identical reagents. Because of highly variable results between laboratories, 7-AAD viability testing of thawed DC needs to be studied further to identify potential causes for the observed variability. |
| doi_str_mv | 10.1080/14653240701744263 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_70224481</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>1_s2_0_S1465324908701531</els_id><sourcerecordid>70224481</sourcerecordid><originalsourceid>FETCH-LOGICAL-c489t-75369507691d1f8631c3f965afabb899bbeb175236b0375711b13bd3ff1e701f3</originalsourceid><addsrcrecordid>eNqFkE1rFTEUhgdR7If-ADcyK3ejOcnkC0EoRa3Q0kV1HTKZE27q3KQmM4XbX28ud6Cg0K4SOM_75uRpmndAPgJR5BP0gjPaE0lA9j0V7EVzDL2UHeVCvNzfBe8qoI-ak1JuCaFEKf66OQJFCdUSjpuzq2Wag8M4Y27LvIy7NsU2xPY-zDm1bmOzdXUWHuwc6iT5dsQ45lBDrcNpKm-aV95OBd-u52nz69vXn-cX3eX19x_nZ5ed65WeO8mZ0JxIoWEErwQDx7wW3Ho7DErrYcABJKdMDIRJLgEGYMPIvAes__PstPlw6L3L6c-CZTbbUPYb2IhpKUYSSvtewbMgJURWX6qCcABdTqVk9OYuh63NOwPE7AWb_wTXzPu1fBm2OD4mVqMV-HwAQvQpb-0G7TRvnM1obtOSY1X0ZP2axmryPmA2xQWMDseQ0c1mTOHJ9Jd_0m4KMTg7_cYdlsf3TaGGmJu1QRNVOzgD9herpq4_</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>20071748</pqid></control><display><type>article</type><title>Multicenter study on in vitro characterization of dendritic cells</title><source>MEDLINE</source><source>Taylor & Francis Journals Complete</source><source>Alma/SFX Local Collection</source><creator>Eichler, H ; Nguyen, X.D ; Roelen, D ; Celluzzi, C.M ; McKenna, D ; Pamphilon, D ; Blair, A ; Read, E.J ; Takahashi, T.A ; Szczepiorkowski, Z.M</creator><creatorcontrib>Eichler, H ; Nguyen, X.D ; Roelen, D ; Celluzzi, C.M ; McKenna, D ; Pamphilon, D ; Blair, A ; Read, E.J ; Takahashi, T.A ; Szczepiorkowski, Z.M ; The Biomedical Excellence for Safer Transfusion (BEST) Collaborative ; Biomedical Excellence for Safer Transfusion Collaborative</creatorcontrib><description>Background There is growing interest in the use of in vitro -expanded dendritic cells (DC) in cancer immunotherapy as cellular-based vaccines. However, the methods used for in vitro preparation vary widely between institutions. Therefore, a strong need exists for standardization, characterization and quality control (QC) of such vaccines. A first prospective multicenter pilot study was performed to investigate basic QC parameters of frozen/thawed DC. The study design was focused on comparison of test results for cell counts, immunophenotyping and cell viability. Methods CD14+ monocytes were isolated from three healthy volunteers. The cells were expanded in vitro , matured and cryopreserved using a standardized protocol in one laboratory. The aliquots of cryopreserved DC and a panel of reagents were shipped to eight laboratories worldwide. The objective was to compare the results of non-functional QC assays between sites by testing identical DC vaccines and using a pre-defined test protocol. Results Measurements of nucleated cell (NC) content of thawed DC vaccines with different types of hematology analyzers (HA) gave similar results for the majority of sites. Immunophenotyping using identical clones of monoclonal antibodies for the detection of surface antigens (i.e. CD1a, CD14, CD16, CD83, CD86 and HLA-DR) provided mostly comparable results between laboratories with an acceptable level of variation. In contrast, highly different results between study sites were generated for measuring the viability of thawed DC by flow cytometry using 7-amino-actinomycin D (7-AAD) dye exclusion. Discussion In characterizing frozen/thawed DC vaccines, NC counts generated by HA yielded similar results between different laboratories. Furthermore, immunophenotyping of DC vaccines can be standardized between centers, i.e. by using identical reagents. Because of highly variable results between laboratories, 7-AAD viability testing of thawed DC needs to be studied further to identify potential causes for the observed variability.</description><identifier>ISSN: 1465-3249</identifier><identifier>EISSN: 1477-2566</identifier><identifier>DOI: 10.1080/14653240701744263</identifier><identifier>PMID: 18202971</identifier><language>eng</language><publisher>England: Elsevier Inc</publisher><subject>Adult ; Advanced Basic Science ; Antibodies, Monoclonal - immunology ; Antigens, CD - immunology ; Antigens, CD1 - immunology ; assay standardization ; B7-2 Antigen - immunology ; CD83 Antigen ; Cell Count ; Cell Survival - immunology ; Cryopreservation - methods ; dendritic cells ; Dendritic Cells - cytology ; Dendritic Cells - immunology ; Flow Cytometry ; good manufacturing practice ; HLA-DR Antigens - immunology ; Humans ; Immunoglobulins - immunology ; Immunophenotyping - methods ; Leukapheresis - methods ; Leukocytes, Mononuclear - cytology ; Leukocytes, Mononuclear - immunology ; Lipopolysaccharide Receptors - immunology ; Male ; Membrane Glycoproteins - immunology ; Middle Aged ; Other ; Pilot Projects ; product characterization ; Prospective Studies ; quality control ; Receptors, IgG - immunology ; vaccines</subject><ispartof>Cytotherapy (Oxford, England), 2008-01, Vol.10 (1), p.21-29</ispartof><rights>International Society for Cellular Therapy</rights><rights>2008 International Society for Cellular Therapy</rights><rights>2008 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted 2008</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c489t-75369507691d1f8631c3f965afabb899bbeb175236b0375711b13bd3ff1e701f3</citedby><cites>FETCH-LOGICAL-c489t-75369507691d1f8631c3f965afabb899bbeb175236b0375711b13bd3ff1e701f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.tandfonline.com/doi/pdf/10.1080/14653240701744263$$EPDF$$P50$$Ginformahealthcare$$H</linktopdf><linktohtml>$$Uhttps://www.tandfonline.com/doi/full/10.1080/14653240701744263$$EHTML$$P50$$Ginformahealthcare$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,61221,61402</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18202971$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Eichler, H</creatorcontrib><creatorcontrib>Nguyen, X.D</creatorcontrib><creatorcontrib>Roelen, D</creatorcontrib><creatorcontrib>Celluzzi, C.M</creatorcontrib><creatorcontrib>McKenna, D</creatorcontrib><creatorcontrib>Pamphilon, D</creatorcontrib><creatorcontrib>Blair, A</creatorcontrib><creatorcontrib>Read, E.J</creatorcontrib><creatorcontrib>Takahashi, T.A</creatorcontrib><creatorcontrib>Szczepiorkowski, Z.M</creatorcontrib><creatorcontrib>The Biomedical Excellence for Safer Transfusion (BEST) Collaborative</creatorcontrib><creatorcontrib>Biomedical Excellence for Safer Transfusion Collaborative</creatorcontrib><title>Multicenter study on in vitro characterization of dendritic cells</title><title>Cytotherapy (Oxford, England)</title><addtitle>Cytotherapy</addtitle><description>Background There is growing interest in the use of in vitro -expanded dendritic cells (DC) in cancer immunotherapy as cellular-based vaccines. However, the methods used for in vitro preparation vary widely between institutions. Therefore, a strong need exists for standardization, characterization and quality control (QC) of such vaccines. A first prospective multicenter pilot study was performed to investigate basic QC parameters of frozen/thawed DC. The study design was focused on comparison of test results for cell counts, immunophenotyping and cell viability. Methods CD14+ monocytes were isolated from three healthy volunteers. The cells were expanded in vitro , matured and cryopreserved using a standardized protocol in one laboratory. The aliquots of cryopreserved DC and a panel of reagents were shipped to eight laboratories worldwide. The objective was to compare the results of non-functional QC assays between sites by testing identical DC vaccines and using a pre-defined test protocol. Results Measurements of nucleated cell (NC) content of thawed DC vaccines with different types of hematology analyzers (HA) gave similar results for the majority of sites. Immunophenotyping using identical clones of monoclonal antibodies for the detection of surface antigens (i.e. CD1a, CD14, CD16, CD83, CD86 and HLA-DR) provided mostly comparable results between laboratories with an acceptable level of variation. In contrast, highly different results between study sites were generated for measuring the viability of thawed DC by flow cytometry using 7-amino-actinomycin D (7-AAD) dye exclusion. Discussion In characterizing frozen/thawed DC vaccines, NC counts generated by HA yielded similar results between different laboratories. Furthermore, immunophenotyping of DC vaccines can be standardized between centers, i.e. by using identical reagents. Because of highly variable results between laboratories, 7-AAD viability testing of thawed DC needs to be studied further to identify potential causes for the observed variability.</description><subject>Adult</subject><subject>Advanced Basic Science</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antigens, CD - immunology</subject><subject>Antigens, CD1 - immunology</subject><subject>assay standardization</subject><subject>B7-2 Antigen - immunology</subject><subject>CD83 Antigen</subject><subject>Cell Count</subject><subject>Cell Survival - immunology</subject><subject>Cryopreservation - methods</subject><subject>dendritic cells</subject><subject>Dendritic Cells - cytology</subject><subject>Dendritic Cells - immunology</subject><subject>Flow Cytometry</subject><subject>good manufacturing practice</subject><subject>HLA-DR Antigens - immunology</subject><subject>Humans</subject><subject>Immunoglobulins - immunology</subject><subject>Immunophenotyping - methods</subject><subject>Leukapheresis - methods</subject><subject>Leukocytes, Mononuclear - cytology</subject><subject>Leukocytes, Mononuclear - immunology</subject><subject>Lipopolysaccharide Receptors - immunology</subject><subject>Male</subject><subject>Membrane Glycoproteins - immunology</subject><subject>Middle Aged</subject><subject>Other</subject><subject>Pilot Projects</subject><subject>product characterization</subject><subject>Prospective Studies</subject><subject>quality control</subject><subject>Receptors, IgG - immunology</subject><subject>vaccines</subject><issn>1465-3249</issn><issn>1477-2566</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1rFTEUhgdR7If-ADcyK3ejOcnkC0EoRa3Q0kV1HTKZE27q3KQmM4XbX28ud6Cg0K4SOM_75uRpmndAPgJR5BP0gjPaE0lA9j0V7EVzDL2UHeVCvNzfBe8qoI-ak1JuCaFEKf66OQJFCdUSjpuzq2Wag8M4Y27LvIy7NsU2xPY-zDm1bmOzdXUWHuwc6iT5dsQ45lBDrcNpKm-aV95OBd-u52nz69vXn-cX3eX19x_nZ5ed65WeO8mZ0JxIoWEErwQDx7wW3Ho7DErrYcABJKdMDIRJLgEGYMPIvAes__PstPlw6L3L6c-CZTbbUPYb2IhpKUYSSvtewbMgJURWX6qCcABdTqVk9OYuh63NOwPE7AWb_wTXzPu1fBm2OD4mVqMV-HwAQvQpb-0G7TRvnM1obtOSY1X0ZP2axmryPmA2xQWMDseQ0c1mTOHJ9Jd_0m4KMTg7_cYdlsf3TaGGmJu1QRNVOzgD9herpq4_</recordid><startdate>20080101</startdate><enddate>20080101</enddate><creator>Eichler, H</creator><creator>Nguyen, X.D</creator><creator>Roelen, D</creator><creator>Celluzzi, C.M</creator><creator>McKenna, D</creator><creator>Pamphilon, D</creator><creator>Blair, A</creator><creator>Read, E.J</creator><creator>Takahashi, T.A</creator><creator>Szczepiorkowski, Z.M</creator><general>Elsevier Inc</general><general>Informa UK Ltd</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>H94</scope><scope>7X8</scope></search><sort><creationdate>20080101</creationdate><title>Multicenter study on in vitro characterization of dendritic cells</title><author>Eichler, H ; Nguyen, X.D ; Roelen, D ; Celluzzi, C.M ; McKenna, D ; Pamphilon, D ; Blair, A ; Read, E.J ; Takahashi, T.A ; Szczepiorkowski, Z.M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c489t-75369507691d1f8631c3f965afabb899bbeb175236b0375711b13bd3ff1e701f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Adult</topic><topic>Advanced Basic Science</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Antigens, CD - immunology</topic><topic>Antigens, CD1 - immunology</topic><topic>assay standardization</topic><topic>B7-2 Antigen - immunology</topic><topic>CD83 Antigen</topic><topic>Cell Count</topic><topic>Cell Survival - immunology</topic><topic>Cryopreservation - methods</topic><topic>dendritic cells</topic><topic>Dendritic Cells - cytology</topic><topic>Dendritic Cells - immunology</topic><topic>Flow Cytometry</topic><topic>good manufacturing practice</topic><topic>HLA-DR Antigens - immunology</topic><topic>Humans</topic><topic>Immunoglobulins - immunology</topic><topic>Immunophenotyping - methods</topic><topic>Leukapheresis - methods</topic><topic>Leukocytes, Mononuclear - cytology</topic><topic>Leukocytes, Mononuclear - immunology</topic><topic>Lipopolysaccharide Receptors - immunology</topic><topic>Male</topic><topic>Membrane Glycoproteins - immunology</topic><topic>Middle Aged</topic><topic>Other</topic><topic>Pilot Projects</topic><topic>product characterization</topic><topic>Prospective Studies</topic><topic>quality control</topic><topic>Receptors, IgG - immunology</topic><topic>vaccines</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Eichler, H</creatorcontrib><creatorcontrib>Nguyen, X.D</creatorcontrib><creatorcontrib>Roelen, D</creatorcontrib><creatorcontrib>Celluzzi, C.M</creatorcontrib><creatorcontrib>McKenna, D</creatorcontrib><creatorcontrib>Pamphilon, D</creatorcontrib><creatorcontrib>Blair, A</creatorcontrib><creatorcontrib>Read, E.J</creatorcontrib><creatorcontrib>Takahashi, T.A</creatorcontrib><creatorcontrib>Szczepiorkowski, Z.M</creatorcontrib><creatorcontrib>The Biomedical Excellence for Safer Transfusion (BEST) Collaborative</creatorcontrib><creatorcontrib>Biomedical Excellence for Safer Transfusion Collaborative</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>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Cytotherapy (Oxford, England)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Eichler, H</au><au>Nguyen, X.D</au><au>Roelen, D</au><au>Celluzzi, C.M</au><au>McKenna, D</au><au>Pamphilon, D</au><au>Blair, A</au><au>Read, E.J</au><au>Takahashi, T.A</au><au>Szczepiorkowski, Z.M</au><aucorp>The Biomedical Excellence for Safer Transfusion (BEST) Collaborative</aucorp><aucorp>Biomedical Excellence for Safer Transfusion Collaborative</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Multicenter study on in vitro characterization of dendritic cells</atitle><jtitle>Cytotherapy (Oxford, England)</jtitle><addtitle>Cytotherapy</addtitle><date>2008-01-01</date><risdate>2008</risdate><volume>10</volume><issue>1</issue><spage>21</spage><epage>29</epage><pages>21-29</pages><issn>1465-3249</issn><eissn>1477-2566</eissn><abstract>Background There is growing interest in the use of in vitro -expanded dendritic cells (DC) in cancer immunotherapy as cellular-based vaccines. However, the methods used for in vitro preparation vary widely between institutions. Therefore, a strong need exists for standardization, characterization and quality control (QC) of such vaccines. A first prospective multicenter pilot study was performed to investigate basic QC parameters of frozen/thawed DC. The study design was focused on comparison of test results for cell counts, immunophenotyping and cell viability. Methods CD14+ monocytes were isolated from three healthy volunteers. The cells were expanded in vitro , matured and cryopreserved using a standardized protocol in one laboratory. The aliquots of cryopreserved DC and a panel of reagents were shipped to eight laboratories worldwide. The objective was to compare the results of non-functional QC assays between sites by testing identical DC vaccines and using a pre-defined test protocol. Results Measurements of nucleated cell (NC) content of thawed DC vaccines with different types of hematology analyzers (HA) gave similar results for the majority of sites. Immunophenotyping using identical clones of monoclonal antibodies for the detection of surface antigens (i.e. CD1a, CD14, CD16, CD83, CD86 and HLA-DR) provided mostly comparable results between laboratories with an acceptable level of variation. In contrast, highly different results between study sites were generated for measuring the viability of thawed DC by flow cytometry using 7-amino-actinomycin D (7-AAD) dye exclusion. Discussion In characterizing frozen/thawed DC vaccines, NC counts generated by HA yielded similar results between different laboratories. Furthermore, immunophenotyping of DC vaccines can be standardized between centers, i.e. by using identical reagents. Because of highly variable results between laboratories, 7-AAD viability testing of thawed DC needs to be studied further to identify potential causes for the observed variability.</abstract><cop>England</cop><pub>Elsevier Inc</pub><pmid>18202971</pmid><doi>10.1080/14653240701744263</doi><tpages>9</tpages></addata></record> |
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| subjects | Adult Advanced Basic Science Antibodies, Monoclonal - immunology Antigens, CD - immunology Antigens, CD1 - immunology assay standardization B7-2 Antigen - immunology CD83 Antigen Cell Count Cell Survival - immunology Cryopreservation - methods dendritic cells Dendritic Cells - cytology Dendritic Cells - immunology Flow Cytometry good manufacturing practice HLA-DR Antigens - immunology Humans Immunoglobulins - immunology Immunophenotyping - methods Leukapheresis - methods Leukocytes, Mononuclear - cytology Leukocytes, Mononuclear - immunology Lipopolysaccharide Receptors - immunology Male Membrane Glycoproteins - immunology Middle Aged Other Pilot Projects product characterization Prospective Studies quality control Receptors, IgG - immunology vaccines |
| title | Multicenter study on in vitro characterization of dendritic cells |
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