Biochemical and Biophysical Research Communications
Inhibition by adiponectin of IL-8 production by human macrophages upon coculturing with late apoptotic cells
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Materials and methods
Cell culture. Human peripheral mononuclear cells were isolated from venous blood of healthy volunteers by Ficoll density gradient centrifugation (Ficoll 400 from Amersham Biosciences). The cells were suspended in RPMI 1640 supplemented with 2% FCS (Invitrogen) at 2.5 × 106 cells/ml and then seeded onto 96-well plates (Corning). After incubation at 37 °C for 1 h, the cells were washed with warmed RPMI 1640 twice and then the medium was replaced by RPMI 1640 supplemented with 10% FCS and 5%
Inhibition by adiponectin of IL-8 production by human macrophages upon coculturing with apoptotic cells
Upon coculturing of human monocyte-derived macrophages with late apoptotic CTLL-2 cells, IL-8 was significantly produced (Fig. 1A, left bar), which was in good agreement with our previous results [12].
We then examined the effect of adiponectin at various concentrations ranging from 3 to 30 μg/ml on IL-8 production by human macrophages upon coculturing with apoptotic cells, because the concentration of adiponectin in human plasma is known to be between 3 and 30 μg/ml [1]. Adiponectin inhibited the
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