Detection of cytokines was carried out using the commercially available BD cytometric bead array (CBA) Mouse Th1/TH2/TH17 cytokine kit (BD, San Jose, CA, USA) as established previously with some minor adaptions (2 (link)). In brief, 10 μL of serum sample was used in 96 well plates and reagent, containing mouse cytokine capture beads with distinct fluorescent intensities, was proportionally adjusted to suit sample size. Samples were measured using a BD Fortessa Flow Cytometer (BD, San Jose, CA, USA) and all sample analysis and data were collected using FlowJo (FlowJo, OR, USA) flow cytometry software.
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