ABSTRACT
Objective: To establish the stable methodology for screening antagonists of CD88, one of C5a receptors. Methods: Whole blood from volunteers was collected and then stimulated by C5a with different concentration for 10-30 min in the presence or absence of PMX-53 and LPS. Flow cytometry was used to detect the expression of CD11b which on the surface of neutrophil. Other functions of nentrophil, including lysozyme release, oxidative burst and interleukin (IL)-8 production, were also examined respectively. Finally, western blotting was used to detect the content of total and phosphorylated ERK and AKT. Thus, the effects about C5a receptor stimulant and the positive medicine PMX-53 on biochemical indicators were investigated. Results: The C5a stimulation actively up-regulated CD11b expression, evoked lysozyme and reactive oxygen species release, increased the IL-8 production and the contents of total and phosphorylated ERK/AKT. While the positive medicine PMX-53 significantly blocked these effects. Conclusion: The methodology in vitro for screening C5a receptor antagonists inhuman whole blood is successfully established.