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For measuring cytokines produced from stimulated PBMCs, 48 h after cultivation in presence of PHA mitogen as described above, supernatants were harvested and kept at −70 °C until testing. IFN-γ (eBioscience, Santa Clara, USA), IL-4 (eBioscience, Santa Clara, USA) and IL-10 (eBioscience, Santa Clara, USA) levels were determined using commercially available ELISA kits according to the manufacturer’s protocol.
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