Analysis of complex biomarkers for human immune-mediated disorders based on cytokine responsiveness of peripheral blood cells.

Davis, J.M. 3rd, Knutson, K.L., Strausbauch, M.A., Crowson, C.S., Therneau, T.M., Wettstein, P.J., Matteson, E.L., Gabriel, S.E.
Journal   J Immunol Methods
Species  
Analytes Measured   G-CSF , GM-CSF , IL-10 , IL-12 Total , IL-13 , IL-2 , IL-4 , IL-5 , IL-6 , IL-7 , IL-8 , MCP-1
Matrix Tested   Peripheral blood mononuclear cell (PBMC) culture supernatants
Year   2010
Volume   184
Page Numbers   7297-7304
Application   Cytokines and Chemokines
Abstract
The advent of improved biomarkers promises to enhance the clinical care for patients with rheumatoid arthritis (RA) and other immune-mediated disorders. We have developed an innovative approach to broadly assess the cytokine responsiveness of human PBMCs using a multistimulant panel and multiplexed immunoassays. The objective of this study was to demonstrate this concept by determining whether cytokine profiles could discriminate RA patients according to disease stage (early versus late) or severity. A 10-cytokine profile, consisting of IL-12, CCL4, TNF-alpha, IL-4, and IL-10 release in response to stimulation with anti-CD3/anti-CD28, CXCL8 and IL-6 in response to CMV and EBV lysate, and IL-17A, GM-CSF, and CCL2 in response to human heat shock protein 60, easily discriminated the early RA group from controls. These data were used to create an immune response score, which performed well in distinguishing the early RA patients from controls and also correlated with several markers of disease severity among the patients with late RA. In contrast, the same 10-cytokine profile assessed in serum was far less effective in discriminating the groups. Thus, our approach lays the foundation for the development of immunologic "signatures" that could be useful in predicting disease course and monitoring the outcomes of therapy among patients with immune-mediated diseases.

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Singleplex
U-PLEX Custom Metabolic Group 1 (human) 384-well Assays
BAFF, BDNF, β-NGF, C-Peptide, CTACK, ENA-78, Eotaxin, Eotaxin-2, Eotaxin-3, EPO, FGF-21, FGF-23, FLT3L, Fractalkine, FSH, G-CSF, Ghrelin (active), Ghrelin (total), GIP (active), GIP (inactive), GIP (total), GLP-1 (active), GLP-1 (inactive), GLP-1 (total), Glucagon, GM-CSF, GRO-α, I-309, IFN-α2a, IFN-β, IFN-γ, IL-1α, IL-1β, IL-1RA, IL-2, IL-2Rα, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12/IL-23p40, IL-12p70, IL-13, IL-15, IL-16, IL-17A, IL-17A/F, IL-17C, IL-17D, IL-17E/IL-25, IL-17F, IL-18, IL-21, IL-22, IL-23, IL-27, IL-29/IFN-λ1, IL-31, IL-33, Insulin, IP-10, Leptin, Luteinizing Hormone (LH), MCP-1, MCP-2, MCP-4, M-CSF, MDC, MIF, MIP-1α, MIP-5, PP, Proinsulin, PYY (total), SDF-1α, TARC, TNF-α, TNF-β, TPO, TRAIL, TSLP, VEGF-A, YKL-40 | Human
Multiplex
U-PLEX Human GM-CSF 384-Well Assay
GM-CSF | Human
Singleplex
U-PLEX Human G-CSF 384-Well Assay
G-CSF | Human
Singleplex
U-PLEX Human IL-13 384-Well Assay
IL-13 | Human
Singleplex
U-PLEX Human IL-10 384-Well Assay
IL-10 | Human
Singleplex
U-PLEX Human IL-2 384-Well Assay
IL-2 | Human
Singleplex
U-PLEX Human MCP-1 384-Well Assay
MCP-1 | Human
Singleplex
U-PLEX Human IL-8 384-Well Assay
IL-8 | Human
Singleplex
U-PLEX Human IL-7 384-Well Assay
IL-7 | Human
Singleplex
U-PLEX Human IL-6 384-Well Assay
IL-6 | Human
Singleplex
U-PLEX Human IL-5 384-Well Assay
IL-5 | Human
Singleplex
U-PLEX Human IL-4 384-Well Assay
IL-4 | Human
Singleplex
U-PLEX Custom Biomarker Group 1 (human) 384-well Assays
CTACK, ENA-78, Eotaxin, Eotaxin-2, Eotaxin-3, EPO, FLT3L, Fractalkine, G-CSF, GM-CSF, GRO-α, I-309, IFN-α2a, IFN-β, IFN-γ, IL-1α, IL-1β, IL-1RA, IL-2, IL-2Rα, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12/IL-23p40, IL-12p70, IL-13, IL-15, IL-16, IL-17A, IL-17A/F, IL-17B, IL-17C, IL-17D, IL-17E/IL-25, IL-17F, IL-18, IL-21, IL-22, IL-23, IL-27, IL-29/IFN-λ1, IL-31, IL-33, IP-10, I-TAC, MCP-1, MCP-2, MCP-3, MCP-4, M-CSF, MDC, MIF, MIP-1α, MIP-1β, MIP-3α, MIP-3β, MIP-5, SDF-1α, TARC, TNF-α, TNF-β, TPO, TRAIL, TSLP, VEGF-A, YKL-40 | Human
Multiplex
S-PLEX Human IL-13 Kit
IL-13 | Human
Singleplex
S-PLEX NHP G-CSF Kit
G-CSF | Non-human primate
Singleplex
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