Critical appraisal of four IL-6 immunoassays.
Abstract
BACKGROUND: Interleukin-6 (IL-6) contributes to numerous inflammatory, metabolic,
and physiologic pathways of disease. We evaluated four IL-6 immunoassays in order
to identify a reliable assay for studies of metabolic and physical function. Serial
plasma samples from intravenous glucose tolerance tests (IVGTTs), with expected rises
in IL-6 concentrations, were used to test the face validity of the various assays.
METHODS AND FINDINGS: IVGTTs, administered to 14 subjects, were performed with a single
infusion of glucose (0.3 g/kg body mass) at time zero, a single infusion of insulin
(0.025 U/kg body mass) at 20 minutes, and frequent blood collection from time zero
to 180 minutes for subsequent Il-6 measurement. The performance metrics of four IL-6
detection methods were compared: Meso Scale Discovery immunoassay (MSD), an Invitrogen
Luminex bead-based multiplex panel (LX), an Invitrogen Ultrasensitive Luminex bead-based
singleplex assay (ULX), and R&D High Sensitivity ELISA (R&D). IL-6 concentrations
measured with MSD, R&D and ULX correlated with each other (Pearson Correlation Coefficients
r = 0.47-0.94, p<0.0001) but only ULX correlated (r = 0.31, p = 0.0027) with Invitrogen
Luminex. MSD, R&D, and ULX, but not LX, detected increases in IL-6 in response to
glucose. All plasma samples were measurable by MSD, while 35%, 1%, and 4.3% of samples
were out of range when measured by LX, ULX, and R&D, respectively. Based on representative
data from the MSD assay, baseline plasma IL-6 (0.90 ± 0.48 pg/mL) increased significantly
as expected by 90 minutes (1.29 ± 0.59 pg/mL, p = 0.049), and continued rising through
3 hours (4.25 ± 3.67 pg/mL, p = 0.0048). CONCLUSION: This study established the face
validity of IL-6 measurement by MSD, R&D, and ULX but not LX, and the superiority
of MSD with respect to dynamic range. Plasma IL-6 concentrations increase in response
to glucose and insulin, consistent with both an early glucose-dependent response (detectable
at 1-2 hours) and a late insulin-dependent response (detectable after 2 hours).
Type
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https://hdl.handle.net/10161/10874Published Version (Please cite this version)
10.1371/journal.pone.0030659Publication Info
Thompson, Dana K; Huffman, Kim M; Kraus, William E; & Kraus, Virginia Byers (2012). Critical appraisal of four IL-6 immunoassays. PLoS One, 7(2). pp. e30659. 10.1371/journal.pone.0030659. Retrieved from https://hdl.handle.net/10161/10874.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Kim Marie Huffman
Associate Professor of Medicine
Determining the role of physical activity in modulating health outcomes (cardiovascular
disease risk) in persons with rheumatologic diseases (rheumatoid arthritis, gout,
osteoarthritis) Integrating clinical rheumatology, basic immunology, metabolism,
and exercise science in order to reduce morbidity in individuals with arthritis Evaluating
relationships between circulating and intra-muscular metabolic intermediates and insulin
resistance in sedentary as well as indiv
Virginia Byers Kraus
Mary Bernheim Distinguished Professor of Medicine
My special area of expertise is as a clinician scientist investigating osteoarthritis.
Osteoarthritis is the most common form of joint disease in man and its incidence increases
with age. It is a problem of increasing concern to the medical community due to the
increasing longevity of the population. Trained as a molecular biologist and a Rheumatologist,
I endeavor to study this disease from bedside to bench. The work in this laboratory
focuses on osteoarthritis and deals w
William Erle Kraus
Richard and Pat Johnson University Distinguished Professor
My training, expertise and research interests range from human integrative physiology
and genetics to animal exercise models to cell culture models of skeletal muscle adaptation
to mechanical stretch. I am trained clinically as an internist and preventive cardiologist,
with particular expertise in preventive cardiology and cardiac rehabilitation. My
research training spans molecular biology and cell culture, molecular genetics, and
integrative human exercise physiology and metabolism. I pr
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