Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT
<p>Abstract</p> <p>Background</p> <p>Cryopreservation of PBMC and/or overnight shipping of samples are required for many clinical trials, despite their potentially adverse effects upon immune monitoring assays such as MHC-peptide tetramer staining, cytokine flow cytomet...
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doaj-30d2f3fe5bae4c58a8130159dfdad5bd2020-11-25T02:10:29ZengBMCBMC Immunology1471-21722005-07-01611710.1186/1471-2172-6-17Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOTMorse Michael AClay Timothy MSummers AmandaChang Jennie CPayne Janice KKuus-Reichel KristineGhanekar Smita AMaino Vernon CBhatia SonnyMoon JamesMaecker Holden TLyerly H KimDeLaRosa CorazonAnkerst Donna PDisis Mary L<p>Abstract</p> <p>Background</p> <p>Cryopreservation of PBMC and/or overnight shipping of samples are required for many clinical trials, despite their potentially adverse effects upon immune monitoring assays such as MHC-peptide tetramer staining, cytokine flow cytometry (CFC), and ELISPOT. In this study, we compared the performance of these assays on leukapheresed PBMC shipped overnight in medium versus cryopreserved PBMC from matched donors.</p> <p>Results</p> <p>Using CMV pp65 peptide pool stimulation or pp65 HLA-A2 tetramer staining, there was significant correlation between shipped and cryopreserved samples for each assay (p ≤ 0.001). The differences in response magnitude between cryopreserved and shipped PBMC specimens were not significant for most antigens and assays. There was significant correlation between CFC and ELISPOT assay using pp65 peptide pool stimulation, in both shipped and cryopreserved samples (p ≤ 0.001). Strong correlation was observed between CFC (using HLA-A2-restricted pp65 peptide stimulation) and tetramer staining (p < 0.001). Roughly similar sensitivity and specificity were observed between the three assays and between shipped and cryopreserved samples for each assay.</p> <p>Conclusion</p> <p>We conclude that all three assays show concordant results on shipped versus cryopreserved specimens, when using a peptide-based readout. The assays are also concordant with each other in pair wise comparisons using equivalent antigen systems.</p> http://www.biomedcentral.com/1471-2172/6/17 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Morse Michael A Clay Timothy M Summers Amanda Chang Jennie C Payne Janice K Kuus-Reichel Kristine Ghanekar Smita A Maino Vernon C Bhatia Sonny Moon James Maecker Holden T Lyerly H Kim DeLaRosa Corazon Ankerst Donna P Disis Mary L |
spellingShingle |
Morse Michael A Clay Timothy M Summers Amanda Chang Jennie C Payne Janice K Kuus-Reichel Kristine Ghanekar Smita A Maino Vernon C Bhatia Sonny Moon James Maecker Holden T Lyerly H Kim DeLaRosa Corazon Ankerst Donna P Disis Mary L Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT BMC Immunology |
author_facet |
Morse Michael A Clay Timothy M Summers Amanda Chang Jennie C Payne Janice K Kuus-Reichel Kristine Ghanekar Smita A Maino Vernon C Bhatia Sonny Moon James Maecker Holden T Lyerly H Kim DeLaRosa Corazon Ankerst Donna P Disis Mary L |
author_sort |
Morse Michael A |
title |
Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT |
title_short |
Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT |
title_full |
Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT |
title_fullStr |
Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT |
title_full_unstemmed |
Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT |
title_sort |
impact of cryopreservation on tetramer, cytokine flow cytometry, and elispot |
publisher |
BMC |
series |
BMC Immunology |
issn |
1471-2172 |
publishDate |
2005-07-01 |
description |
<p>Abstract</p> <p>Background</p> <p>Cryopreservation of PBMC and/or overnight shipping of samples are required for many clinical trials, despite their potentially adverse effects upon immune monitoring assays such as MHC-peptide tetramer staining, cytokine flow cytometry (CFC), and ELISPOT. In this study, we compared the performance of these assays on leukapheresed PBMC shipped overnight in medium versus cryopreserved PBMC from matched donors.</p> <p>Results</p> <p>Using CMV pp65 peptide pool stimulation or pp65 HLA-A2 tetramer staining, there was significant correlation between shipped and cryopreserved samples for each assay (p ≤ 0.001). The differences in response magnitude between cryopreserved and shipped PBMC specimens were not significant for most antigens and assays. There was significant correlation between CFC and ELISPOT assay using pp65 peptide pool stimulation, in both shipped and cryopreserved samples (p ≤ 0.001). Strong correlation was observed between CFC (using HLA-A2-restricted pp65 peptide stimulation) and tetramer staining (p < 0.001). Roughly similar sensitivity and specificity were observed between the three assays and between shipped and cryopreserved samples for each assay.</p> <p>Conclusion</p> <p>We conclude that all three assays show concordant results on shipped versus cryopreserved specimens, when using a peptide-based readout. The assays are also concordant with each other in pair wise comparisons using equivalent antigen systems.</p> |
url |
http://www.biomedcentral.com/1471-2172/6/17 |
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