Bacterial DNA and endothelial damage in haemodialysis patients

BACKGROUND: An increased percentage of CD14+CD16+ activated monocytes have been reported in peripheral blood from haemodialysis patients. The aim of this study is to investigate if a mild stimulus such as bacterial DNA (CpG-ODNs) contamination may induce an inflammatory response in CD14+CD16+ monocytes from haemodialysis patients and to value the biological consequences of this inflammatory response on endothelial cell damage. METHODS: Circulating mononuclear cells from 20 haemodialysis patients and 15 healthy subjects were studied. CD14+CD16+ and the toll-like receptor 9 (TLR-9) expression were assessed by flow cytometry. Cell culture inserts were used to evaluate the effect of CD14+CD16+ and CpG-ODNs on endothelial cell apoptosis (measured by Tunnel). Intracellular cytokines were measured by Cytometric methods. NF-kappaB, p38 MAPK, c-Jun PI3K and MEK1/2 activity were modified by specific peptides. RESULTS: At baseline, CD14+CD16+ have an increased expression of cytoquines and TLR-9. CpG-ODNs caused the production and release of cytoquines in CD14+CD16+, but not in CD14++ monocytes. This inflammatory response was mediated by intracellular signalling dependent on NF-kappaB, p38 MARK or c-Jun PI3K but not by MEK1/2 activation. The results of the present study also demonstrate that the inflammatory response induced by the stimulation of CD14+CD16+ by CpG DNA resulted in endothelial cell apoptosis. CONCLUSIONS: The results of the present study demonstrate that in haemodialysis patients there is a subpopulation of pre-activated monocytes that can be stimulated by contaminant bacterial DNA. These activated cells produce and release inflammatory factors that may cause endothelial injury.