G-CSF application in patients with severe bacterial pneumonia increases IL-10 expression in neutrophils

Abstract 

In severe pneumonia, the application of granulocyte-colony stimulating factor (G-CSF) was associated with reduced complications possibly by an induction of anti-inflammatory cytokines. It is not clear, whether G-CSF induces interleukin-10 (IL-10) synthesis in neutrophils. In a randomized study, 15 patients with severe community acquired pneumonia were treated either by a single dose of G-CSF and antibiotic therapy (n=8) or antibiotics alone (n=7). Messenger ribonucleic acid (mRNA) expression of IL-10 and tumor necrosis factor alpha of peripheral blood leukocytes was measured using in-situ hybridization (ISH) and reverse-transcription-polymerase-chain-reaction (RT-PCR). In addition, the cytokine release of lipopolysaccharide (LPS)-stimulated whole blood was measured by ELISA. We detected increased IL-10 mRNA by ISH (140±8% vs. −11±5%, P<0.01) and RT-PCR (126±16% vs. −28±3%, P<0.01) in the G-CSF-treated group only. In contrast, LPS-stimulated whole blood cellsin vitro released significantly less IL-10 compared to the control group (−38.2±9.7 vs. −14.8±6 pg/ml, P<0.02). There was no significant effect on IL-10 serum protein levels and the TNF-α release and expression. IL-10 mRNA was detected predominantly in cluster designation 66b (CD66b) positive nucleated blood cells indicating that polymorphonuclear leukocytes are the main source of IL-10 expression after G-CSF stimulation. G-CSF induces transcription of IL-10 mRNA in neutrophils without increased release. This may be due to posttranscriptional effects.

Keywords: G-CSF, pneumonia, IL-10, neutrophils, in-situ hybridization.

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• f1 Correspondence should be addressed to: Dr S. Spuck, Klinik fuer Neurochirurgie, Medizinische Universitaet zu Luebeck, Ratzeburger Allee 160, D-23538 Luebeck, Germany; E-mail: s.spuck@freenet.de