TY - JOUR
T1 - Multiplex measurement of proinflammatory cytokines in human serum
T2 - Comparison of the Meso Scale Discovery electrochemiluminescence assay and the Cytometric Bead Array
AU - Dabitao, Djeneba
AU - Margolick, Joseph B.
AU - Lopez, Joseph
AU - Bream, Jay H.
N1 - Funding Information:
This work was supported by (1) the HIV Prevention Trials Network (HPTN) sponsored by the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Child Health and Human Development (NICH/HD), National Institute on Drug Abuse, National Institute of Mental Health, and Office of AIDS Research, of the NIH, DHHS ( 1U01AI068613 ), (2) UL1-RR025005 ( GCRC ), and (3) U01AI35042 ( SHARE ). We thank Stacey Meyerer and Akila Hadji for specimen management, and Jacquett R. Johnson and Lisette M. Johnson-Hill for data management. We also thank the Becton Dickinson Immune Function Laboratory in the Johns Hopkins Bloomberg School of Public Health for access to instrumentation and software.
Funding Information:
The authors thank the Division of Clinical Research (DCR) of the NIAID, NIH, DHHS and the NIAID/University of Bamako HIV and Tuberculosis Research and Training Center (SEREFO Project) in Mali for their financial support to Djeneba Dabitao.
PY - 2011/9/30
Y1 - 2011/9/30
N2 - Serum cytokine profiling is a powerful tool to link host immune defense with disease pathogenesis. Although several multiplex assays are commercially available, none has been rigorously validated in the context of chronic infectious disease (such as HIV infection). Here we compared the measurement of proinflammatory cytokines by two multiplex platforms: the Meso Scale Discovery (MSD) electrochemiluminescence assay and the Becton Dickinson Cytometric Bead Array (CBA) flow cytometric assay, using serum samples from HIV-infected and -uninfected donors. We evaluated the ability of these assays to: a) quantify circulating levels of native cytokines (IL-6, IL-8, IL-10, TNF-α, IL-12p70, IL-1β), and b) accurately recover known amounts of recombinant cytokines added to serum samples. Based on the standard curves, the sensitivity of the MSD system was only slightly better than the CBA. However, in serum the MSD platform consistently quantified levels of endogenous IL-12p70, TNF-α, and IL-10 that were undetectable by the CBA assay. The MSD assay was also more accurate as determined by an enhanced capacity to recover known concentrations of recombinant cytokines added to serum. Both assays performed equally well in quantifying IL-6 and IL-8, while neither assay quantified IL-1β with accuracy and precision. Interestingly, HIV infection did not affect the performance of either assay. Overall, the MSD assay provided a more reliable assessment of the proinflammatory cytokines tested in the serum of healthy and HIV-infected individuals.
AB - Serum cytokine profiling is a powerful tool to link host immune defense with disease pathogenesis. Although several multiplex assays are commercially available, none has been rigorously validated in the context of chronic infectious disease (such as HIV infection). Here we compared the measurement of proinflammatory cytokines by two multiplex platforms: the Meso Scale Discovery (MSD) electrochemiluminescence assay and the Becton Dickinson Cytometric Bead Array (CBA) flow cytometric assay, using serum samples from HIV-infected and -uninfected donors. We evaluated the ability of these assays to: a) quantify circulating levels of native cytokines (IL-6, IL-8, IL-10, TNF-α, IL-12p70, IL-1β), and b) accurately recover known amounts of recombinant cytokines added to serum samples. Based on the standard curves, the sensitivity of the MSD system was only slightly better than the CBA. However, in serum the MSD platform consistently quantified levels of endogenous IL-12p70, TNF-α, and IL-10 that were undetectable by the CBA assay. The MSD assay was also more accurate as determined by an enhanced capacity to recover known concentrations of recombinant cytokines added to serum. Both assays performed equally well in quantifying IL-6 and IL-8, while neither assay quantified IL-1β with accuracy and precision. Interestingly, HIV infection did not affect the performance of either assay. Overall, the MSD assay provided a more reliable assessment of the proinflammatory cytokines tested in the serum of healthy and HIV-infected individuals.
KW - Biomarker
KW - Cytokine
KW - HIV
KW - Multiplex assay
KW - Serum
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U2 - 10.1016/j.jim.2011.06.033
DO - 10.1016/j.jim.2011.06.033
M3 - Article
C2 - 21781970
AN - SCOPUS:80052495503
SN - 0022-1759
VL - 372
SP - 71
EP - 77
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -