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Title:
VACUTAINER® CPT™ and Ficoll density gradient separation perform equivalently in maintaining the quality and function of PBMC from HIV seropositive blood samples | BMC Immunology
Description:
Background For immune monitoring studies during HIV vaccine clinical trials, whole blood specimens from HIV seropositive (HIV+) patients may be collected at multiple sites and sent to a central location for peripheral blood mononuclear cell (PBMC) isolation, cryopreservation and functional evaluation. In this study we show a comparison of two PBMC preparation options, Ficoll density gradient separation (Ficoll) and Cell Preparation Tubes (CPT) using shipped whole blood specimens from 19 HIV+ patients (CD4 > 350, viral load < 50). The pre- and post- cryopreservation performance of samples collected by these two methods were compared by assessment of antigen-specific IFNγ expression in CD8+ and CD8- T cells, cellular viability, and cellular recovery. Results The results indicate that cryopreserved PBMC samples tested for CMV- and HIV- specific interferon-gamma (IFNγ) expression performed equivalent to the respective fresh PBMC processed under both collection conditions. Compared to fresh PBMC, the viability was significantly lower for cryopreserved PBMC derived using Ficoll, although it was never less than 90%. There were no significant differences in the IFNγ response, viability, or recovery between cryopreserved PBMC derived by Ficoll and by CPT. Conclusion These data suggest that CPT is an efficient system for the collection and cryopreservation of functionally active HIV+ PBMC, as well as a viable alternative to Ficoll gradient separation.
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Keywords {🔍}
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Topics {✒️}
paired t-test antigen-specific memory/effector cd4+ hiv-p55 peptide mix hiv-p55 peptide mixes cell preparation tube evacuation type/vacutainer® tubes hiv- specific interferon-gamma cmv-pp65 peptide mix cmv-pp65 peptide mix cell preparation tubes article download pdf multicenter clinical trials polypropylene v-bottom plate assessing antigen-specific responses labor-intensive process requiring cryopreserved mononuclear cells antigen-specific homeostatic mechanism cytokine flow cytometry cpt-processing routinely yields clinical trials antigen-specific ifnγ expression single-cell assays cell subsets cell cytokine expression diminished cell numbers ficoll-processed cryopreserved pbmc ficoll gradient separation flow cytometry data antigen-specific lymphocyte function full access cryopreserved ficoll-processed samples cpt-processed pbmc demonstrated privacy choices/manage cookies article-processing charge cryopreserved cpt-processed samples authors’ original file cmvpp65 peptide mix hivp55 peptide mix hiv-infected patients cell separation ficoll-processed fresh pbmc median post-thaw recovery fresh ficoll-processed samples cpt-processed fresh pbmc 19 hiv-seropositive donors pbmc preparation options clinical researchers prefer immune monitoring studies pre-determined time points cell responses
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headline:VACUTAINER® CPT™ and Ficoll density gradient separation perform equivalently in maintaining the quality and function of PBMC from HIV seropositive blood samples
description:For immune monitoring studies during HIV vaccine clinical trials, whole blood specimens from HIV seropositive (HIV+) patients may be collected at multiple sites and sent to a central location for peripheral blood mononuclear cell (PBMC) isolation, cryopreservation and functional evaluation. In this study we show a comparison of two PBMC preparation options, Ficoll density gradient separation (Ficoll) and Cell Preparation Tubes (CPT) using shipped whole blood specimens from 19 HIV+ patients (CD4 > 350, viral load < 50). The pre- and post- cryopreservation performance of samples collected by these two methods were compared by assessment of antigen-specific IFNγ expression in CD8+ and CD8- T cells, cellular viability, and cellular recovery. The results indicate that cryopreserved PBMC samples tested for CMV- and HIV- specific interferon-gamma (IFNγ) expression performed equivalent to the respective fresh PBMC processed under both collection conditions. Compared to fresh PBMC, the viability was significantly lower for cryopreserved PBMC derived using Ficoll, although it was never less than 90%. There were no significant differences in the IFNγ response, viability, or recovery between cryopreserved PBMC derived by Ficoll and by CPT. These data suggest that CPT is an efficient system for the collection and cryopreservation of functionally active HIV+ PBMC, as well as a viable alternative to Ficoll gradient separation.
datePublished:2006-05-25T00:00:00Z
dateModified:2006-05-25T00:00:00Z
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Peripheral Blood Mononuclear Cell
Good Viability
Peripheral Blood Mononuclear Cell Sample
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Cell Preparation Tube
Immunology
Allergology
Vaccine
Cytokines and Growth Factors
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headline:VACUTAINER® CPT™ and Ficoll density gradient separation perform equivalently in maintaining the quality and function of PBMC from HIV seropositive blood samples
description:For immune monitoring studies during HIV vaccine clinical trials, whole blood specimens from HIV seropositive (HIV+) patients may be collected at multiple sites and sent to a central location for peripheral blood mononuclear cell (PBMC) isolation, cryopreservation and functional evaluation. In this study we show a comparison of two PBMC preparation options, Ficoll density gradient separation (Ficoll) and Cell Preparation Tubes (CPT) using shipped whole blood specimens from 19 HIV+ patients (CD4 > 350, viral load < 50). The pre- and post- cryopreservation performance of samples collected by these two methods were compared by assessment of antigen-specific IFNγ expression in CD8+ and CD8- T cells, cellular viability, and cellular recovery. The results indicate that cryopreserved PBMC samples tested for CMV- and HIV- specific interferon-gamma (IFNγ) expression performed equivalent to the respective fresh PBMC processed under both collection conditions. Compared to fresh PBMC, the viability was significantly lower for cryopreserved PBMC derived using Ficoll, although it was never less than 90%. There were no significant differences in the IFNγ response, viability, or recovery between cryopreserved PBMC derived by Ficoll and by CPT. These data suggest that CPT is an efficient system for the collection and cryopreservation of functionally active HIV+ PBMC, as well as a viable alternative to Ficoll gradient separation.
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Good Viability
Peripheral Blood Mononuclear Cell Sample
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Cell Preparation Tube
Immunology
Allergology
Vaccine
Cytokines and Growth Factors
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