Tumor necrosis factor measurement and use of different anticoagulants: Possible interference in plasma samples and supernatants from endotoxin-stimulated monocytes

Hoffmann, J. N.; Hartl, Wolfgang H.; Faist, Eugen; Jochum, Marianne; Inthorn, D.

doi:10.1007/s000110050199

Cited by 20 publications

(11 citation statements)

References 29 publications

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“…EDTA and citrate both remove calcium to prevent coagulation while heparin acts instead by binding to antithrombin III[12]. Indeed, we observed only two differences in cytokine measurements between EDTA and citrate, while more differences were noted when comparing heparin to both EDTA and citrate.…”

Section: Discussionmentioning

confidence: 68%

Effect of anticoagulants on multiplexed measurement of cytokine/chemokines in healthy subjects

et al. 2012

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Introduction Cytokines are humoral regulatory molecules that act together in immunologic pathways. Monitoring cytokines and their variations within physiologic ranges is critical for biomarker discovery. Therefore, we evaluated the performance characteristics of 72 analytes measured by multiplex cytokine immunoassay, with an emphasis on the differences of analytes measured in serum compared to plasma, and, in plasma, on the impact of anticoagulants on the cytokine measurement. Methods We used fluorescent bead-based (Luminex) immunoassay kits to simultaneously measure 72 analytes. We tested serum and plasma samples from 11 matched donors. Plasma samples were anti-coagulated with sodium heparin, sodium citrate dextrose and ethylene diamine tetra-acetic acid (EDTA), respectively. Results Of the 72 cytokines, 12 were undetectable in all types of specimen samples. Nineteen analytes, including PDGF-bb, IL-4, IL-8, IL-9, FGF-b, PAI-1, CXCL-5, CCL-5, CD40L, EGF, VEGF, IL-2ra, IL-3, SDF-1a, PCT, MCP-3, GIP, IL-16 and fibrinogen, showed significant differences between measurements in serum and all types of plasma, regardless of anticoagulant. Among plasma samples, 10 analytes (eotaxin, SCGF-b, MCP-1, SCF, MIP-1b, VEGF, RANTES, PDGF-b, PAI-1 and ITAC) showed significantly higher concentrations in heparinized plasma compared to citrated and EDTA plasma. IP-10, and CTAK were the only 2 cytokines that presented different concentrations in citrate and EDTA plasma. Conclusions With their small volume, low cost per test, and multiplex capacity, Luminex-based cytokine assays have enormous potential utility for screening in epidemiologic studies. In our study, we showed that many cytokines’ concentrations differed between serum and plasma samples, and that different anticoagulants used in preparation of plasma samples also affected the measurement of some cytokines. There was no optimal sample preparation that was clearly superior for the measurement of all analytes measured. Ultimately, the utility of cytokine measurement, as biomarker or to monitor the immune system, will depend on attention to detail in the collection and processing of samples in addition to assay precision.

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Section: Discussionmentioning

confidence: 68%

Effect of anticoagulants on multiplexed measurement of cytokine/chemokines in healthy subjects

et al. 2012

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“…This phenomenon was not identified for serum samples, suggesting that dilution of the anticoagulant could play a role in the decrease in recovery values that has been reported elsewhere. 8,13,14 The manufacturer of the multiplex assay system evaluated in the present study recommends use of EDTA-treated plasma or serum as the test solution; use of heparin as an anticoagulant had not been previ- (Table 1). See Figure 1 for remainder of key.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a feline-specific multiplex, bead-based assay for detection of cytokines, chemokines, growth factors, and other immunologically active proteins in serum and plasma samples from cats

Halpin¹,

Saunders²,

Thompson³

et al. 2016

ajvr

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OBJECTIVE To evaluate a feline-specific multiplex, bead-based assay system for detection of recombinant and native proteins in serum samples and in EDTA-treated and heparinized plasma samples. SAMPLE Serum samples and EDTA-treated and heparinized plasma samples from 30 sick cats and 9 healthy client-owned cats and heparinized whole blood samples from 5 healthy purpose-bred cats. PROCEDURES Ability of the assay system to detect 19 recombinant and native immunologically active proteins in plasma and serum samples from healthy and purpose-bred cats was evaluated via spike-and-recovery tests, assessments of inter- and intra-assay variation, linearity results, and leukocyte stimulation. Effects of various concentrations of heparin and serum matrix solution on percentages of analytes recovered were also evaluated. Analyte concentrations in samples from healthy and sick cats were measured and compared between groups. RESULTS Percentages of analytes recovered were unsatisfactory for most assays. Serum and heparinized plasma samples yielded better recovery results than did EDTA-treated plasma samples. Use of serum matrix solution did not improve results. Use of heparin concentrations greater than the recommended range affected the results. Linearity of results was difficult to assess because of the poor recovery. For the analytes that were recovered sufficiently for assessment, linearity appeared to be reasonable despite the limited detection. CONCLUSIONS AND CLINICAL RELEVANCE Poor percentages of analytes recovered and adverse effects of sample protein matrix limited the usefulness of the multiplex, bead-based assay system for measurement of immunologically active proteins in solutions with high protein content; however, recovery results were fairly linear, potentially allowing evaluation of feline plasma or serum samples with high analyte concentrations.

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“…To obtain plasma, various anticoagulants can be used (ethylenediaminetetraacetic acid (EDTA), lithium/sodium heparin, and sodium citrate). Cytokine measurements were shown to be affected by the anticoagulant used [78] and, notably, lithium heparin and sodium citrate were shown to affect levels of IL-6 and TNF-alpha compared to EDTA plasma [35, 112, 113]. Citrate plasma collection also results in the reduction of total protein concentration due to the volume of citrate anticoagulant diluting the blood, in addition to an osmotic withdrawal of water from blood cells [114].…”

Section: Cytokines As Biomarkersmentioning

confidence: 99%

Cytokines as Biomarkers in Rheumatoid Arthritis

Burska

Boissinot

Ponchel

2014

Mediators of Inflammation

179

139

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RA is a complex disease that develops as a series of events often referred to as disease continuum. RA would benefit from novel biomarker development for diagnosis where new biomarkers are still needed (even if progresses have been made with the inclusion of ACPA into the ACR/EULAR 2010 diagnostic criteria) and for prognostic notably in at risk of evolution patients with autoantibody-positive arthralgia. Risk biomarkers for rapid evolution or cardiovascular complications are also highly desirable. Monitoring biomarkers would be useful in predicting relapse. Finally, predictive biomarkers for therapy outcome would allow tailoring therapy to the individual. Increasing numbers of cytokines have been involved in RA pathology. Many have the potential as biomarkers in RA especially as their clinical utility is already established in other diseases and could be easily transferable to rheumatology. We will review the current knowledge's relation to cytokine used as biomarker in RA. However, given the complexity and heterogeneous nature of RA, it is unlikely that a single cytokine may provide sufficient discrimination; therefore multiple biomarker signatures may represent more realistic approach for the future of personalised medicine in RA.

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Tumor necrosis factor measurement and use of different anticoagulants: Possible interference in plasma samples and supernatants from endotoxin-stimulated monocytes

Cited by 20 publications

References 29 publications

Effect of anticoagulants on multiplexed measurement of cytokine/chemokines in healthy subjects

Effect of anticoagulants on multiplexed measurement of cytokine/chemokines in healthy subjects

Evaluation of a feline-specific multiplex, bead-based assay for detection of cytokines, chemokines, growth factors, and other immunologically active proteins in serum and plasma samples from cats

Cytokines as Biomarkers in Rheumatoid Arthritis

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