title: Optimization of storage and shipment of cryopreserved peripheral blood mononuclear cells from HIV-infected and uninfected individuals for ELISPOT assays.
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%.
authors.affiliation: Infectious Diseases, University of Colorado Denver, Aurora, CO, United States. adriana.weinberg@ucdenver.edu
authors.forename: Adriana
authors.initials: A
authors.lastname: Weinberg
authors.forename: Lin Ye
authors.initials: LY
authors.lastname: Song
authors.forename: Cindy L
authors.initials: CL
authors.lastname: Wilkening
authors.forename: Terry
authors.initials: T
authors.lastname: Fenton
authors.forename: John
authors.initials: J
authors.lastname: Hural
authors.forename: Raul
authors.initials: R
authors.lastname: Louzao
authors.forename: Guido
authors.initials: G
authors.lastname: Ferrari
authors.forename: Paige E
authors.initials: PE
authors.lastname: Etter
authors.forename: Mark
authors.initials: M
authors.lastname: Berrong
authors.forename: Jennifer D
authors.initials: JD
authors.lastname: Canniff
authors.forename: Donald
authors.initials: D
authors.lastname: Carter
authors.forename: Olivier D
authors.initials: OD
authors.lastname: Defawe
authors.forename: Ambrosia
authors.initials: A
authors.lastname: Garcia
authors.forename: Tania L
authors.initials: TL
authors.lastname: Garrelts
authors.forename: Rebecca
authors.initials: R
authors.lastname: Gelman
authors.forename: Linda K
authors.initials: LK
authors.lastname: Lambrecht
authors.forename: Savita
authors.initials: S
authors.lastname: Pahwa
authors.forename: Sudheesh
authors.initials: S
authors.lastname: Pilakka-Kanthikeel
authors.forename: David L
authors.initials: DL
authors.lastname: Shugarts
authors.forename: Nancy B
authors.initials: NB
authors.lastname: Tustin
mesh_terms: D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D005260:Female; D015658:HIV Infections; D006801:Humans; D007963:Leukocytes, Mononuclear; D008297:Male; D013997:Time Factors