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Assays of neutrophil cytokine release in vitro
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To stimulate cytokine release, 1 × 106 freshly-isolated peripheral neutrophils were incubated with LPS (2 μg/mL) in RPMI 1640 with 1% BSA for 18 h. The level of cytokine release in the cell-free supernatants was determined using ELISA kits for TNF-α (LifeSpan, F12799) and IL-1β (Abbkine, KET9001) according to the manufacturers’ instructions.
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