Expression data from primary human neutrophils incubated with complement anaphylatoxin C5a

Influence of complement anaphylatoxin C5a on neutrophil gene expression Trauma causes an early activation of the complement system, which leads to excessive generation of the anaphylatoxin C5a. Furthermoren, alterations in neutrophil function are often associated with infectious complication after trauma. Studies have shown that C5a is a main contributor to neutrophil dysfunction. However, the pathophysiological mechanisms still remain elusive. Aim of the study was to evaluate whether C5a can induce changes in expression profiles. We identified distinct classes of up-regulated genes during this process. Primary polymorhoneuclear neutrophils (PMNs) were isolated from human blood drawn by peripheral venous puncture in sodium citrate monovettes. PMNs were isolated from whole blood by Ficoll-Hypaque gradient centrifugation, dextran sedimentation, followed by hypotonic lysis of residual red blood cells. CD9+ eosinophils were separated from neutrophils. Purified Neutrophils were either left untreated (n=3) or incubated with native human C5a (100ng/ml) for 60 min at 37°C on a rotating wheel (n=3).