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BMC Immunology
Published in: BMC Immunology 1/2005

Open Access 01-12-2005 | Methodology article

Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT

Authors: Holden T Maecker, James Moon, Sonny Bhatia, Smita A Ghanekar, Vernon C Maino, Janice K Payne, Kristine Kuus-Reichel, Jennie C Chang, Amanda Summers, Timothy M Clay, Michael A Morse, H Kim Lyerly, Corazon DeLaRosa, Donna P Ankerst, Mary L Disis

Published in: BMC Immunology | Issue 1/2005

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Abstract

Background

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.

Results

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.

Conclusion

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.
Appendix
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Metadata
Title
Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT
Authors
Holden T Maecker
James Moon
Sonny Bhatia
Smita A Ghanekar
Vernon C Maino
Janice K Payne
Kristine Kuus-Reichel
Jennie C Chang
Amanda Summers
Timothy M Clay
Michael A Morse
H Kim Lyerly
Corazon DeLaRosa
Donna P Ankerst
Mary L Disis
Publication date
01-12-2005
Publisher
BioMed Central
Published in
BMC Immunology / Issue 1/2005
Electronic ISSN: 1471-2172
DOI
https://doi.org/10.1186/1471-2172-6-17

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