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

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

doi:10.1186/1471-2172-6-17

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

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

Research output: Contribution to journal › Article › peer-review

110 Scopus citations

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.

Original language	English
Article number	17
Journal	BMC Immunology
Volume	6
DOIs	https://doi.org/10.1186/1471-2172-6-17
State	Published - 18 Jul 2005
Externally published	Yes

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10.1186/1471-2172-6-17

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Maecker, H. T., Moon, J., Bhatia, S., Ghanekar, S. A., Maino, V. C., Payne, J. K., Kuus-Reichel, K., Chang, J. C., Summers, A., Clay, T. M., Morse, M. A., Lyerly, H. K., DeLaRosa, C., Ankerst, D. P., & Disis, M. L. (2005). Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT. BMC Immunology, 6, Article 17. https://doi.org/10.1186/1471-2172-6-17

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title = "Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT",

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.",

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

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doi = "10.1186/1471-2172-6-17",

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T1 - Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT

AU - Maecker, Holden T.

AU - Moon, James

AU - Bhatia, Sonny

AU - Ghanekar, Smita A.

AU - Maino, Vernon C.

AU - Payne, Janice K.

AU - Kuus-Reichel, Kristine

AU - Chang, Jennie C.

AU - Summers, Amanda

AU - Clay, Timothy M.

AU - Morse, Michael A.

AU - Lyerly, H. Kim

AU - DeLaRosa, Corazon

AU - Ankerst, Donna P.

AU - Disis, Mary L.

PY - 2005/7/18

Y1 - 2005/7/18

N2 - 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.

AB - 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.

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U2 - 10.1186/1471-2172-6-17

DO - 10.1186/1471-2172-6-17

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SN - 1471-2172

VL - 6

JO - BMC Immunology

JF - BMC Immunology

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ER -

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

Abstract

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