Cytokine and Chemokine Profiling in COVID-19

Plasma samples were centrifuged for 15 min at 1,500 × g followed by dilution at 2- and 4-fold for quantifying cytokine and chemokine profiles, respectively. Concentrations of cytokines and chemokines were quantified using the V-plex Plus proinflammatory panel 1, cytokine panel 1 kit, and chemokine panel 1 kits from Meso Scale Discovery (MSD) according to the manufacturer’s instruction, which included IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, TNF-α, GM-CSF, IL-1α, IL-5, IL-7, IL-12/23p40, IL-15, IL-16, IL-17A, TNF-α, VEGF, eotaxin, MIP-1α, eotaxin-3, TARC, IP-10, MIP-1β, MCP-1, MDC, and MCP-4. A total of 120 plasma samples from COVID-19 patients and 59 plasma samples from healthy subjects were examined for cytokine/chemokine analysis. Data were acquired on the V-plex Sector Imager 2400 plate reader. Analyte concentrations were extrapolated from a standard curve calculated using a four-parameter logistic fit using MSD Workbench 3.0 software. CRP concentrations were received from the patient’s clinical files. The plasma Gal-9 was quantified using ELISA (R&D; DY 2045) as we reported elsewhere (24 (link), 25 (link)).