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Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
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.
Clinical and Vaccine Immunology : CVI
|June 24, 2011
Summary
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.
