Cytokine production
MLRs were applied in the combinations as described above. On day 5,
supernatants from each well (100 μL) of selected samples (n = 13) were
collected and frozen. The concentration of cytokines in each supernatant
sample was analyzed in batches after thawing using a custom designed
Luminex plate as per the manufacturer’s instructions (Bio-Techne,
Minneapolis, MI, USA). The concentration of IFN-γ, IL-4, IL-6, IL-8,
IL-10, IL-17 and TNF-α were analyzed using a Bio-Plex 200 plate reader
and the Bio-Plex Manager 6.1 software (Bio-Rad, Hercules, CA, USA).