Optimization of storage and shipment of cryopreserved peripheral blood mononuclear cells from HIV-infected and uninfected individuals for ELISPOT assays.
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%.
Weinberg
Adriana
A
Infectious Diseases, University of Colorado Denver, Aurora, CO, United States. adriana.weinberg@ucdenver.edu
Song
Lin Ye
LY
Wilkening
Cindy L
CL
Fenton
Terry
T
Hural
John
J
Louzao
Raul
R
Ferrari
Guido
G
Etter
Paige E
PE
Berrong
Mark
M
Canniff
Jennifer D
JD
Carter
Donald
D
Defawe
Olivier D
OD
Garcia
Ambrosia
A
Garrelts
Tania L
TL
Gelman
Rebecca
R
Lambrecht
Linda K
LK
Pahwa
Savita
S
Pilakka-Kanthikeel
Sudheesh
S
Shugarts
David L
DL
Tustin
Nancy B
NB
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