Citrated whole blood was prepared by centrifugation at 2500 gfor 15 min at RT. Then, 1 ml plasma was collected in a 1.5-ml Eppendorf tube and
centrifuged at 10,000 g for 10 min at 4℃. The top supernatant
(800 µl) was collected prior to freezing, and stored at –80℃. Measurements of
cytokines were performed using a microsphere-based multiplexing bioassay system
with Xmap technology (Austin, TX, USA) with a Luminex IS 100 instrument
(Bio-Rad, Hercules, CA, USA), powered with the Bio-Plex manager Software version
6.0 (build 617). The anti-inflammatory cytokine IL-10, the MyD88-dependent
cytokines IL-1β, IL-6, IL-8 and TNF-α,13 and the MyD88-independent
cytokines IP-10 and RANTES,15 (link) were measured (Bio-Rad, Oslo, Norway) in duplicates.
centrifuged at 10,000 g for 10 min at 4℃. The top supernatant
(800 µl) was collected prior to freezing, and stored at –80℃. Measurements of
cytokines were performed using a microsphere-based multiplexing bioassay system
with Xmap technology (Austin, TX, USA) with a Luminex IS 100 instrument
(Bio-Rad, Hercules, CA, USA), powered with the Bio-Plex manager Software version
6.0 (build 617). The anti-inflammatory cytokine IL-10, the MyD88-dependent
cytokines IL-1β, IL-6, IL-8 and TNF-α,13 and the MyD88-independent
cytokines IP-10 and RANTES,15 (link) were measured (Bio-Rad, Oslo, Norway) in duplicates.
