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http://hdl.handle.net/123456789/1860
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| Title: | Quality monitoring of HIV-1-infected and uninfected peripheral blood mononuclear cell samples in a resource-limited setting |
| Authors: | Olemukam, Robert E.
Eller, Leigh Anne
Ouma, Benson J.
Etonu, Ben
Erima, Simon
Naluyima, Prossy
Kyabaggu, Denis
Cox, Josephine H.
Sandberg, Johan K.
Wabwire-Mangen, Fred
Michael, Nelson L.
Robb, Merlin L.
de Souza, Mark S.
Eller, Michael A. |
| Keywords: | Human immunodeficiency virus type 1
HIV-1
peripheral blood mononuclear cell
PBMC
Cryopreservation |
| Issue Date: | Jun-2010 |
| Publisher: | American Society for Microbiology |
| Citation: | Olemukam, R. E. et al. (2010). Quality monitoring of HIV-1-infected and uninfected peripheral blood mononuclear cell samples in a resource-limited setting. Clinical and Vaccine Immunology, 17(6): 910–918 |
| Abstract: | Human immunodeficiency virus type 1 (HIV-1) vaccine and natural history studies are critically dependent
on the ability to isolate, cryopreserve, and thaw peripheral blood mononuclear cell (PBMC) samples with a
high level of quality and reproducibility. Here we characterize the yield, viability, phenotype, and function of
PBMC from HIV-1-infected and uninfected Ugandans and describe measures to ascertain reproducibility and
sample quality at the sites that perform cryopreservation. We have developed a comprehensive internal quality
control program to monitor processing, including components of method validation. Quality indicators for
real-time performance assessment included the time from venipuncture to cryopreservation, time for PBMC
processing, yield of PBMC from whole blood, and viability of the PBMC before cryopreservation. Immune
phenotype analysis indicated lowered B-cell frequencies following processing and cryopreservation for both
HIV-1-infected and uninfected subjects (P < 0.007), but all other major lymphocyte subsets were unchanged.
Long-term cryopreservation did not impact function, as unstimulated specimens exhibited low background and
all specimens responded to staphylococcal enterotoxin B (SEB) by gamma interferon and interleukin-2
production, as measured by intracellular cytokine staining. Samples stored for more than 3 years did not decay
with regard to yield or viability, regardless of HIV-1 infection status. These results demonstrate that it is
possible to achieve the high level of quality necessary for vaccine trials and natural history studies in a
resource-limited setting and provide strategies for laboratories to monitor PBMC processing performance. |
| URI: | http://dx.doi.org/10.1128/CVI.00492-09
http://hdl.handle.net/123456789/1860 |
| ISSN: | 1556-6811 |
| Appears in Collections: | Research Articles (Health-Sciences)
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Files in This Item:
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| Olemukam-chs-res.pdf | | 1109Kb | Adobe PDF | View/Open |
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