Performance evaluation of human cytokines profiles obtained by various multiplexed-based technologies underlines a need for standardization

Dupuy, Anne‐Marie; Kuster, Nils; Lizard, Gérard; Ragot, Kévin; Lehmann, Sylvain; Gallix, B.; Cristol, Jean Paul

doi:10.1515/cclm-2012-0648

Cited by 22 publications

(16 citation statements)

References 12 publications

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“…Indeed, we confirmed that the absolute values of the proteins detected were different between the two tested technologies. Such discordance could be due to differences in the capture and/or detection antibodies and to a differential sensitivity to matrix interference (Dupuy et al, 2013;Khan et al, 2004). However, among the cytokines for which the quantification was consistent, we observed strong inter-platform correlations for IL-8 and VEGF (Table 5), in accordance with previous findings (Chowdhury et al, 2009;Dabitao et al, 2011;Huttunen et al, 2014).…”

Section: Discussionsupporting

confidence: 89%

“…Indeed, both platforms failed to robustly detect the GM-CSF, IL-1β, IL-2 and IL-5 cytokines for which previous investigations had reported significant variations in expression either during MDE treatment or at baseline in comparison to healthy controls (Biancotto et al, 2013;Dahl et al, 2014;Hernandez et al, 2008;Schmidt et al, 2014;Shelton et al, 2015). These types of inconsistency in the results have already been reported (Breen et al, 2011;Chaturvedi et al, 2011;Dabitao et al, 2011;Dupuy et al, 2013;Fu et al, 2010). In particular, the consensus that the detection of IL-1β suffers from poor performances is of importance.…”

Section: Discussionmentioning

confidence: 94%

“…Nevertheless, it has been shown that (i) both serum and plasma inhibit the readouts, (ii) the inhibitory effect of the matrix is partly reversed by dilution, and (iii) this type of inhibition is much higher with the Luminex® system compared to the MSD assays (Rosenberg-Hasson et al, 2014). In addition, as previously noted in a few studies (Dossus et al, 2009;Dupuy et al, 2013;Toedter et al, 2008), the nature of the capture/detection antibody reactivity is crucial. In fact, for some cytokines such as IL-4, IL12p70 and IL-17 the Bio-Plex assays invariably outperformed those of the V-PLEX in both sera and plasma, whereas for other analytes including bFGF, GM-CSF and IL-15, striking differences in the detectability between sera and plasma were observed.…”

Section: Discussionmentioning

confidence: 97%

See 2 more Smart Citations

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

Belzeaux

Lefebvre

Lazzari

et al. 2017

Psychoneuroendocrinology

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Cytokines produced by both immune and non-immune cells are likely to play roles in the development and/or progression of psychiatric disorders. Indeed, many investigators have compared the blood cytokine levels in psychiatric patients with those of healthy controls or monitored their levels in patients during disease progression to identify biomarkers. Nevertheless, very few studies have confirmed that such cytokines remain stable in healthy individuals through periods of weeks and months. This is an important issue to consider before using blood cytokine levels as biomarkers of disease traits, disease state, or treatment response. Although multiplex assay technology represents an advance in identifying biomarkers because it allows simultaneous examination of large panels of analytes from a small volume of sample, it is necessary to verify whether these assays yield enough sensitivity and reproducibility when applied to the blood from neuropsychiatric patients. Therefore, we compared two multiplex immunoassays, the bead-based Luminex® (Bio-Rad) and the electro-chemiluminescence-based V-plex® (MesoScaleDiscovery), for the detection and quantification of 31 cytokines, chemokines and growth factors in both the sera and plasma of patients with major depressive episodes (MDE) and age-and sex-matched healthy control subjects during a 30-week period. Although both platforms exhibited low coefficients of variability (CV) between the duplicates in the calibration curves, the linearity was better in general for the V-PLEX® platform. However, neither platform was able to detect the absolute values for all of the tested analytes. Among the 16 analytes that were detected by both assays, the intra-assay reproducibility was in general better with the V-PLEX® platform. Although it is not a general rule that the results from sera and plasma will be correlated, consistent results were more frequent with the V-PLEX® platform. Furthermore, the V-PLEX® results were more consistent with the gold standard ELISA simplex assay for IL-6 in both sera and plasma. The intra-individual variability of the measurements, among the sera and plasma for the 4 samples harvested from each healthy individual, was low for Eotaxin, G-CSF, IL-4, IL-7, IL-9, IL-12p40, IL-12p70, IL-15, MIP-1β, PDGF-BB, TNF, TNF-β and VEGF, but intermediate or high for IFN-γ, IL-6, IL-8, IL-10, and IP10. Together, these data suggest that extreme caution is needed in translating the results of multiplex cytokine profiling into biomarker discovery in psychiatry. Dear Editor,We thank you and the Editor in chief, Robert Dantzer, for reaching the conclusion to ultimately publish our manuscript. Following your suggestion, we changed the tiltle to: "How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays".We were concerned by the persistent negative comments of Reviewer#3, stressing out that our comparison between the two platforms we have worked with suffers a number of theoretical and technical limitations because...

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 97%

See 1 more Smart Citation

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

Belzeaux

Lefebvre

Lazzari

et al. 2017

Psychoneuroendocrinology

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“…However, analytical performances were adequate for longitudinal studies using any 1 method, but standardization was required before results obtained with different methods could be compared. 45 Because comparisons between various commercially available cytokine BBMAs are not completely similar, it will be important to include positive controls (high, medium, low), dilute standards within the given sample matrix, and validate the standards prior to performing the assay on large-scale experimental studies, checking for linearity, recovery, repeatability, and obtaining a dynamic range that is acceptable for the samples to be tested.…”

mentioning

confidence: 99%

Opportunities for bead-based multiplex assays in veterinary diagnostic laboratories

et al. 2013

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Abstract. Bead-based multiplex assays (BBMAs) are applicable for high throughput, simultaneous detection of multiple analytes in solution (from several to 50-500 analytes within a single, small sample volume). Currently, few assays are commercially available for veterinary applications, but they are available to identify and measure various cytokines, growth factors and their receptors, inflammatory proteins, kinases and inhibitors, neurobiology proteins, and pathogens and antibodies in human beings, nonhuman primates, and rodent species. In veterinary medicine, various nucleic acid and protein-coupled beads can be used in, or for the development of, antigen and antibody BBMAs, with the advantage that more data can be collected using approximately the same amount of labor as used for other antigen and antibody assays. Veterinary-related BBMAs could be used for detection of pathogens, genotyping, measurement of hormone levels, and in disease surveillance and vaccine assessment. It will be important to evaluate whether BBMAs are "fit for purpose," how costs and efficiencies compare between assays, which assays are published or commercially available for specific veterinary applications, and what procedures are involved in the development of the assays. It is expected that many veterinary-related BBMAs will be published and/or become commercially available in the next few years. The current review summarizes the BBMA technology and some of the currently available BBMAs developed for veterinary settings. Some of the human diagnostic BBMAs are also described, providing an example of possible templates for future development of new veterinary-related BBMAs.

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“…It should be noted that most of the standard deviations are very high, suggesting a skewed distribution, with a few patients with very high levels and a non-negligible number of patients with cytokine concentrations below the limit of detection [1,2]. This high number of nondetected data (.50%) has been recently described for cytokines assessed by three commercially available multiplex assays [7]. Consequently, caution should be considered about low cytokine circulating concentrations, because most probably measured with an analytical imprecision .20%, making such promising clinical findings somewhat questionable.…”

mentioning

confidence: 70%

Cytokines in pulmonary arterial hypertension: consider sensitivity when using multiplex technology

Monneret¹

2014

Eur Respir J

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Cytokines in pulmonary arterial hypertension: consider sensitivity when using multiplex technology To the Editor:CRACOWSKI et al. [1] have recently published an interesting substudy in which they analysed the association of 17 baseline plasma cytokine concentrations with mortality in patients suffering from pulmonary arterial hypertension (PAH), using multiplex technology. The authors showed that four cytokines were independently associated with an increase in the adjusted hazard of mortality. Previously, SOON et al. [2] demonstrated that elevated levels of cytokines in PAH also predict survival over a 5-year period. Both these studies reinforce the increasing evidence that inflammation is associated with PAH remodelling.Nevertheless, it is known that concentrations of circulating cytokines are very low, in the pg?mL -1 range, requiring highly sensitive methods. In the letter by CRACOWSKI et al.[1], despite a well-defined design and a robust statistical analysis, as previously described [3], no information is provided about the analytical performances of the method for cytokine quantification. In order to strengthen such cytokine-based clinical studies, the sensitivity of multiplex technology is herein discussed.

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Performance evaluation of human cytokines profiles obtained by various multiplexed-based technologies underlines a need for standardization

Cited by 22 publications

References 12 publications

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

How to: Measuring blood cytokines in biological psychiatry using commercially available multiplex immunoassays

Opportunities for bead-based multiplex assays in veterinary diagnostic laboratories

Cytokines in pulmonary arterial hypertension: consider sensitivity when using multiplex technology

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