Optimization of storage and shipment of cryopreserved peripheral blood mononuclear cells from HIV-infected and uninfected individuals for ELISPOT assays

Adriana Weinberg, Lin Ye Song, Cindy L. Wilkening, Terry Fenton, John Hural, Raul Louzao, Guido Ferrari, Paige E. Etter, Mark Berrong, Jennifer D. Canniff, Donald Carter, Olivier D. Defawe, Ambrosia Garcia, Tania L. Garrelts, Rebecca Gelman, Linda K. Lambrecht, Savita Pahwa, Sudheesh Pilakka-Kanthikeel, David L. Shugarts, Nancy B. Tustin

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

Functional immunologic assays using cryopreserved peripheral blood mononuclear cells (PBMC) are influenced by blood processing, storage and shipment. The objective of this study was to compare the viability, recovery and ELISPOT results of PBMC stored and shipped in liquid nitrogen (LN/LN) or stored in LN and shipped on dry ice (LN/DI) or stored at -70 °C for 3 to 12. weeks and shipped on DI (70/DI 3 to 12); and to assess the effect of donor HIV infection status on the interaction between storage/shipment and the outcome measures. PBMC from 12 HIV-infected and 12 uninfected donors showed that LN/LN conferred higher viability and recovery than LN/DI or 70/DI 3, 6, 9 or 12. LN/DI PBMC had higher viability than any 70/DI PBMC. The PBMC viability and recovery linearly decreased with the duration of storage at -70 °C from 3 to 12. weeks. This effect was more pronounced in samples from HIV-infected than uninfected donors. Results of ELISPOT assays using CMV pp65, CEF and Candida albicans antigens were qualitatively and quantitatively similar across LN/LN, LN/DI and 70/DI 3. However, ELISPOT values significantly decreased with the duration of storage at -70 °C both in HIV-infected and uninfected donors. ELISPOT results also decreased with PBMC viability < 70%.

Original languageEnglish (US)
Pages (from-to)42-50
Number of pages9
JournalJournal of Immunological Methods
Volume363
Issue number1
DOIs
StatePublished - Dec 15 2010

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Keywords

  • ELISPOT
  • HIV
  • PBMC
  • Viable cryopreservation

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Weinberg, A., Song, L. Y., Wilkening, C. L., Fenton, T., Hural, J., Louzao, R., Ferrari, G., Etter, P. E., Berrong, M., Canniff, J. D., Carter, D., Defawe, O. D., Garcia, A., Garrelts, T. L., Gelman, R., Lambrecht, L. K., Pahwa, S., Pilakka-Kanthikeel, S., Shugarts, D. L., & Tustin, N. B. (2010). Optimization of storage and shipment of cryopreserved peripheral blood mononuclear cells from HIV-infected and uninfected individuals for ELISPOT assays. Journal of Immunological Methods, 363(1), 42-50. https://doi.org/10.1016/j.jim.2010.09.032