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

BMC Immunology - Tập 6 Số 1 - 2005
Holden T. Maecker1, James Moon2, Sonny Bhatia1, Smita Ghanekar1, Vernon C. Maino1, Janice K. Payne3, Kristine Kuus-Reichel3, Juan Chang3, Amanda Summers4, Timothy M. Clay4, Michael A. Morse4, H. Kim Lyerly4, Corazon delaRosa5, Donna P. Ankerst2, Mary L. Disis5
1BD Biosciences, San Jose, USA
2Southwest Oncology Group Statistical Center at Fred Hutchinson Cancer Research Center, Seattle, USA
3Beckman-Coulter, San Diego, USA
4Departments of Surgery, Medicine, Pathology, and Immunology, and Duke Comprehensive Cancer Center, Duke University Medical Center, Durham, USA
5Tumor Vaccine Group, Division of Oncology, University of Washington, Seattle, USA

Tóm tắt

AbstractBackgroundCryopreservation 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.ResultsUsing 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.ConclusionWe 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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