Figure 6. Probenecid but not pannexin-1 blockers reduce P2X7-induced IL-1β secretion from human monocytes and J774 macrophages.

Magnetically isolated CD14⁺ human monocytes were primed with LPS (100 ng/ml) for 4 hours and stimulated with 3 mM ATP for 15 minutes. After stimulation supernatants were removed and tested for IL-1β. (A) representative data from one donor, (B) shows pooled data from 6 donors expressed as % of ATP control. Carbenoxolone (CBX) was used at 50 μM, probenecid (PRO) at 1 mM and 2.5 mM, ¹⁰Panx1 at 100 μM, and A-438079 at 10 μM. (C) The J774 mouse macrophage cell line was primed with LPS (1 μg/ml) and stimulated with ATP (5 mM) for 20 minutes in physiological saline. P2X7 selective antagonist AZ10606120 (AZ106) was used at 10 μM. Mean data from three independent experiments is shown. ATP-induced IL-1β released into supernatant was measured by specific ELISAs for human and mouse IL-1β respectively. Error bars represent S.E.M and * represents P<0.05.