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

被引：50

作者：

Weinberg, Adriana ^{[1
]}

Song, Lin Ye ^{[2
]}

Wilkening, Cindy L. ^{[2
]}

Fenton, Terry ^{[2
]}

Hural, John ^{[3
]}

Louzao, Raul ^{[4
]}

Ferrari, Guido ^{[5
]}

Etter, Paige E. ^{[6
]}

Berrong, Mark ^{[5
]}

Canniff, Jennifer D. ^{[1
]}

Carter, Donald ^{[3
]}

Defawe, Olivier D. ^{[3
]}

Garcia, Ambrosia ^{[4
]}

Garrelts, Tania L. ^{[4
]}

Gelman, Rebecca ^{[2
]}

Lambrecht, Linda K. ^{[7
]}

Pahwa, Savita ^{[8
]}

Pilakka-Kanthikeel, Sudheesh ^{[8
]}

Shugarts, David L. ^{[1
]}

Tustin, Nancy B. ^{[9
]}

机构：

[1] Univ Colorado Denver, Aurora, CO USA

[2] Harvard Univ, Sch Publ Hlth, Ctr Biostat AIDS Res, Boston, MA 02115 USA

[3] Fred Hutchinson Canc Res Ctr, HIV Vaccine Trials Network, Seattle, WA 98104 USA

[4] Duke Univ, Duke Human Vaccine Inst, Durham, NC USA

[5] Duke Univ, Med Ctr, Dept Surg, Durham, NC 27710 USA

[6] HIV AIDS Network Coordinat HANC, Seattle, WA USA

[7] Univ Massachusetts, Worcester, MA 01605 USA

[8] Univ Miami, Miller Sch Med, Dept Microbiol & Immunol, Dev Ctr AIDS Res DCFAR, Miami, FL 33136 USA

[9] Childrens Hosp Philadelphia, Res Inst, Philadelphia, PA 19104 USA

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2010年 / 363卷 / 01期

关键词：

PBMC; HIV; ELISPOT; Viable cryopreservation; ANTIRETROVIRAL THERAPY; IMMUNE-RESPONSES; VACCINE; CHILDREN; TRIALS;

D O I：

10.1016/j.jim.2010.09.032

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

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 degrees 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 degrees 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 degrees C both in HIV-infected and uninfected donors. ELISPOT results also decreased with PBMC viability <70%. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：42 / 50

页数：9

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