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Cytokine analysis

DS Deepti Singh
PG Paul C. Guest
HD Henrik Dobrowolny
TF Tino Fischbach
GM Gabriela Meyer-Lotz
CB Carolin Breitling-Ziegler
AH Aiden Haghikia
SV Stefan Vielhaber
JS Johann Steiner
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Concentrations of 13 cytokines [interferon (IFN)-α2, IFN-γ, interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IL-18, IL-23, IL-33, monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-α] were determined in triplicate using the LEGENDplex™ Human Inflammation Panel 1 (BioVendor; Brno, Czechia) according to manufacturer instructions. This is a fluorescence–coded microsphere-based multiplex immuoassay for the detection and quantitation of analytes by flow cytometry. Sera were diluted twofold and CSF tested undiluted. Standard curves of each cytokine were analysed in duplicate on two separate occasions. The LOB and LOD were determined for each assay according to CLSI guidelines [16, 22], using the formulas below:

For statistical analysis, we used only those cytokines for which at least 2 of the 3 replicates and > 50% of samples gave readings above the LOD.

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