Recombinant human C5a induces transcription but not translation of interleukin 1 beta mRNA in human monocytes.

The effect of recombinant human C5a (rhC5a) on the synthesis of interleukin 1 beta (IL-1 beta) was investigated in human monocytes, isolated by leukapheresis and countercurrent elutriation. rhC5a induced IL-1 beta mRNA synthesis in a dose- and time-dependent manner. Maximal induction was achieved at 3 h with rhC5a concentrations of 200 to 500 ng/ml. The maximal rhC5a-stimulated mRNA induction was about 75% of that observed using LPS as the stimulus (300 ng/ml). The inducing activity of rhC5a could be neutralized by preincubation with a polyclonal anti-C5a antibody. On the other hand rhC5a in optimal concentrations only weakly stimulated IL-1 beta protein synthesis, as measured by a two-site directed enzyme-linked immunoassay. When compared on Northern blots, a slightly reduced mobility of C5a-stimulated IL-1 beta mRNA was observed relative to LPS-stimulated RNA. To exclude the possibility that structural defects in the IL-1 beta mRNA are responsible for the weak translational efficiency after C5a stimulation a primer extension experiment was performed. No difference in the length of the extended fragment was detected between LPS and C5a-stimulated RNA, suggesting that the 5'-regions of the RNAs are identical. When LPS- or C5a-stimulated RNA was used to program IL-1 beta synthesis in an in vitro translation system from rabbit reticulocytes, no difference in translational efficiency was observed. Our results indicate that in human monocytes two signals for IL-1 beta gene expression are necessary, a signal for transcriptional activation and another signal to induce translation of the mRNA.