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Diagnostic Laboratory Immunology

Value of a Quality Assessment Program in Optimizing Cryopreservation of Peripheral Blood Mononuclear Cells in a Multicenter Study

Najib Aziz, Joseph B. Margolick, Roger Detels, Charles R. Rinaldo, John Phair, Beth D. Jamieson, Anthony W. Butch
Najib Aziz
aDavid Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA
bFielding School of Public Health, University of California, Los Angeles, Los Angeles, California, USA
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Joseph B. Margolick
cJohns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA
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Roger Detels
aDavid Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA
bFielding School of Public Health, University of California, Los Angeles, Los Angeles, California, USA
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Charles R. Rinaldo
dGraduate School of Public Health, University of Pittsburgh, Pittsburgh, Pennsylvania, USA
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John Phair
eFeinberg School of Medicine, Northwestern University, Chicago, Illinois, USA
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Beth D. Jamieson
aDavid Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA
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Anthony W. Butch
aDavid Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA
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DOI: 10.1128/CVI.00693-12
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    Fig 1

    Viability and recovery of cryopreserved PBMC from 75 HIV-uninfected and 76 HIV-infected participants across the four MACS sites. The solid lines represent the median, and the dashed lines represent the mean. The box represents the 25th to 75th percentiles. The lower and upper horizontal bars represent the 10th and 90th percentiles, respectively.

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    Fig 2

    Viable cell recovery from each vial of cryopreserved PBMC from 75 HIV-uninfected and 76 HIV-infected participants. Numbers on the x axis represent sequential times when cryopreserved PBMC from the four MACS sites were tested. The QA testing program was started in July 2004 and was performed every 4 months. The first symbol in each testing round for each of the MACS sites is the value for the HIV-uninfected participants, and the second symbol is the value for the HIV-infected participants. The dashed lines represent 75% and 100% viable cell recoveries.

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    Fig 3

    Proliferative responses of cryopreserved PBMC (n = 28 per group) to phytohemagglutinin (PHA). The stimulation index was defined as the counts per minute in the presence of PHA divided by the counts per minute in the absence of PHA. The solid lines represent medians, and the dashed lines represent the means of the distributions. For HIV-uninfected samples the median and mean were the same. The boxes represent the 25th to 75th percentiles. The lower and upper horizontal bars indicate the 10th and 90th percentiles, respectively.

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    Fig 4

    Correlation between the PHA-induced proliferative response of cryopreserved PBMC and the percentage of CD4+ (top panel) and CD8+ (bottom panel) T lymphocytes for 27 HIV-uninfected and 27 HIV-infected participants. Least-squares regression lines are indicated.

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    Fig 5

    Deming regression analyses of expression of cell surface markers CD3 (top panel), CD4 (middle panel), and CD8 (bottom panel) on paired samples of fresh whole-blood and cryopreserved PBMC. A total of 54 paired blood samples (27 from HIV-uninfected and 27 from HIV-infected participants) from the 4 MACS sites were evaluated. The dashed lines represent the line of identity (y = x), and the solid lines represent the least-squares regression lines of the data.

Tables

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  • Table 1

    T lymphocyte phenotypes in fresh whole-blood and cryopreserved PBMC

    Percentage typeCD3+ (%)CD3+ CD4+ (%)CD3+ CD8+ (%)
    FreshCryopreservedFreshCryopreservedFreshCryopreserved
    Mean767635374038
    Minimum474010101818
    Maximum899263707677
    Median767836383936
    IQRa10.110.131.331.325.123.2
    • ↵a IQR is the interquartile range, the difference between the 25th and 75th percentiles.

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Value of a Quality Assessment Program in Optimizing Cryopreservation of Peripheral Blood Mononuclear Cells in a Multicenter Study
Najib Aziz, Joseph B. Margolick, Roger Detels, Charles R. Rinaldo, John Phair, Beth D. Jamieson, Anthony W. Butch
Clinical and Vaccine Immunology Mar 2013, 20 (4) 590-595; DOI: 10.1128/CVI.00693-12

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Value of a Quality Assessment Program in Optimizing Cryopreservation of Peripheral Blood Mononuclear Cells in a Multicenter Study
Najib Aziz, Joseph B. Margolick, Roger Detels, Charles R. Rinaldo, John Phair, Beth D. Jamieson, Anthony W. Butch
Clinical and Vaccine Immunology Mar 2013, 20 (4) 590-595; DOI: 10.1128/CVI.00693-12
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