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Published on: November 9, 2017
Multisite comparison of high-sensitivity multiplex cytokine assays
Elizabeth Crabb Breen1, Sandra M Reynolds, Christopher Cox
1UCLA Cousins Center for Psychoneuroimmunology, Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA 90095-7076, USA.
Insights
High-sensitivity multiplex cytokine assays show promise for immune status monitoring, but variability exists between labs and lots. These assays may be best for tracking relative cytokine changes in longitudinal HIV studies.
Area of Science:
- Immunology
- Biotechnology
- Clinical Diagnostics
Background:
- Cytokine concentrations in serum and plasma indicate immune status.
- Low cytokine levels necessitate high-sensitivity assays for detection.
- Multiplex assays enable simultaneous measurement of multiple cytokines from small samples, ideal for limited volumes.
Purpose of the Study:
- Evaluate four high-sensitivity multiplex cytokine assays for detecting 13 cytokines in human serum and plasma.
- Assess laboratory and lot variability across different multiplex platforms (Luminex, Meso Scale Discovery).
- Determine the utility of these assays for detecting cytokine changes during primary HIV viremia.
Main Methods:
- Four high-sensitivity multiplex cytokine assays were tested on Luminex and electrochemiluminescence platforms.
- Archived serum from HIV-uninfected and -infected subjects (MACS, WIHS) and commercial plasma samples were used.
- Assays were performed across six different laboratories to evaluate reproducibility.
Main Results:
- Interleukin-6 (IL-6), IL-8, IL-10, and tumor necrosis factor alpha were detectable by most kits.
- Interleukin-1 beta (IL-1β) was detected by only one kit.
- Significant differences (P < 0.001) in results were observed between laboratories and/or lots for all kits.
- Despite variability, assays generally detected similar patterns of cytokine changes during primary HIV viremia.
Conclusions:
- Current multiplex cytokine assays exhibit variability in detecting serum/plasma concentrations.
- Reproducibility issues may limit their use in long-term, multi-laboratory studies requiring absolute values.
- These assays can be valuable for longitudinal studies focusing on relative cytokine changes.
Abstract:
The concentrations of cytokines in human serum and plasma can provide valuable information about in vivo immune status, but low concentrations often require high-sensitivity assays to permit detection. The recent development of multiplex assays, which can measure multiple cytokines in one small sample, holds great promise, especially for studies in which limited volumes of stored serum or plasma are available. Four high-sensitivity cytokine multiplex assays on a Luminex (Bio-Rad, BioSource, Linco) or electrochemiluminescence (Meso Scale Discovery) platform were evaluated for their ability to detect circulating concentrations of 13 cytokines, as well as for laboratory and lot variability. Assays were performed in six different laboratories utilizing archived serum from HIV-uninfected and -infected subjects from the Multicenter AIDS Cohort Study (MACS) and the Women's Interagency HIV Study (WIHS) and commercial plasma samples spanning initial HIV viremia. In a majority of serum samples, interleukin-6 (IL-6), IL-8, IL-10, and tumor necrosis factor alpha were detectable with at least three kits, while IL-1β was clearly detected with only one kit. No single multiplex panel detected all cytokines, and there were highly significant differences (P < 0.001) between laboratories and/or lots with all kits. Nevertheless, the kits generally detected similar patterns of cytokine perturbation during primary HIV viremia. This multisite comparison suggests that current multiplex assays vary in their ability to measure serum and/or plasma concentrations of cytokines and may not be sufficiently reproducible for repeated determinations over a long-term study or in multiple laboratories but may be useful for longitudinal studies in which relative, rather than absolute, changes in cytokines are important.
