4.5. Luminex Assays

Twenty-nine cytokine and chemokine species, including epidermal growth factor (EGF), eotaxin, granulocyte macrophage-colony stimulating factor (GM-CSF), G-CSF, interferon-alpha2 (IFN-α), IFN-γ, interleukin 1 alpha (IL-1α), IL-1β, IL-1 receptor antagonist (IL-1RA), IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p40, IL-12p70, IL-13, IL-15, IL-17a, IFN-γ-inducible protein-10 (IP-10), monocyte chemotactic protein 1 (MCP-1), macrophage-inducible protein 1α (MIP-1α), MIP-1β, tumor necrosis factor α (TNF-α), TNF-β, and vascular endothelial growth factor (VEGF), in plasma were measured using a commercially available kit (Milliplex Human Cytokine and Chemokine multiplex assay kit, Merck Millipore, Billerica, MA, USA) by Luminex methods, as reported previously [17]. The assay was performed according to manufacturer′s instructions and the concentrations of cytokines/chemokines were calculated by comparing the assay readings with a five-parameter logistic standard curve on a Bioplex-200 instrument (Bio-Rad, Hercules, CA, USA). All of the results were expressed in pg/mL.