Resolvin D5 on prostaglandin E2 mediated inhibition of fMLF induced activation of the neutrophil NADPH
Description
The data show the effects on the inhibitory activity of PGE2 in the presence/absense of the allosterically modelator RvD5, when neutrophils were activated by the FPR1 agonist fMLF. Neutrophils were isolated from peripheral blood of healthy adults obtained from the blood bank at Sahlgrenska University Hospital and the release of ROS by activted neutrophils was measured using a very senstitive isoluminol-amplified chemiluminescence system.
Files
Steps to reproduce
Human blood neutrophils isolated from buffy coats were incubated with PGE2 in the presence or absence of the allosteric modulator resolvin RvD5. The neutrophilds were then activated by the FPR1 agonist fMLF and receptor downstream function was monitored as production of superoxide anions. The O2− production was measured in 6-channel Biolumat LB 9505 (Berthold Co, Wildbad, Germany), using disposable 4-mL polypropylene tubes with a 0.9 mL reaction mixture. Neutrophils (1 × 105), HRP (4 U/mL), isoluminol (2 × 10−5 M) and optimal antagonist/inhibitor (KRG for control) were incubated (five min 37 °C) before stimulation with 0.1 mL agonist and the release of O2−was measured as light emission over time. Data are shown with a representative chemiluminescence kinetics, abscissa time (min) and ordinate chemiluminescence arbitrary units (Mega counts per minute, Mcpm).
Institutions
- Goteborgs Universitet