M Heesen1, B Blömeke, B Schlüter, N Heussen, R Rossaint, D Kunz. 1. Department of Anesthesiology and Intensive Care Medicine, University Hospital of Aachen, Pauwelsstrasse 30, 52057 Aachen, Germany. M.Heesen@amc.uva.nl
Abstract
OBJECTIVE: CD14 is a receptor for endotoxin and binds components of Gram-positive and Gram-negative bacteria. CD14-bearing monocytes respond to stimulation with the increased synthesis and release of cytokines. The recently described -260 C-->T promoter polymorphism of the CD14 gene has been found to be related to a risk of myocardial infarction. This study evaluated the role of this polymorphism in the expression of monocyte and soluble CD14. Moreover, the effect of the CD14 -260 genotypes for the ex vivo TNF-alpha response to endotoxin was analyzed in whole blood. PATIENTS AND PARTICIPANTS: Ninety-five healthy blood donors were studied. MEASUREMENTS AND RESULTS: CD14 -260 genotyping was performed by means of a real-time PCR with fluorescence labeled hybridization probes. CD14 expression on human monocytes (mCD14) was assessed by fluorescence-activated cell sorting analysis with anti-CD14 monoclonal antibodies. Plasma levels of soluble CD14 (sCD14) were measured by ELISA. The TNF-alpha synthesis was determined by chemiluminescence in whole blood after endotoxin stimulation. There were no differences in mCD14 density, sCD14 levels, or the tumor necrosis factor-alpha concentrations between individuals with the three different CD14 -260 genotypes CC, CT, and TT. CONCLUSIONS: The CD14 -260 polymorphism does not affect the CD14 expression of unstimulated circulating monocytes or soluble CD14 plasma levels.
OBJECTIVE:CD14 is a receptor for endotoxin and binds components of Gram-positive and Gram-negative bacteria. CD14-bearing monocytes respond to stimulation with the increased synthesis and release of cytokines. The recently described -260 C-->T promoter polymorphism of the CD14 gene has been found to be related to a risk of myocardial infarction. This study evaluated the role of this polymorphism in the expression of monocyte and soluble CD14. Moreover, the effect of the CD14 -260 genotypes for the ex vivo TNF-alpha response to endotoxin was analyzed in whole blood. PATIENTS AND PARTICIPANTS: Ninety-five healthy blood donors were studied. MEASUREMENTS AND RESULTS:CD14 -260 genotyping was performed by means of a real-time PCR with fluorescence labeled hybridization probes. CD14 expression on human monocytes (mCD14) was assessed by fluorescence-activated cell sorting analysis with anti-CD14 monoclonal antibodies. Plasma levels of soluble CD14 (sCD14) were measured by ELISA. The TNF-alpha synthesis was determined by chemiluminescence in whole blood after endotoxin stimulation. There were no differences in mCD14 density, sCD14 levels, or the tumor necrosis factor-alpha concentrations between individuals with the three different CD14 -260 genotypes CC, CT, and TT. CONCLUSIONS: The CD14 -260 polymorphism does not affect the CD14 expression of unstimulated circulating monocytes or soluble CD14 plasma levels.
Authors: Alex P Reiner; Ethan M Lange; Nancy S Jenny; Paulo H M Chaves; Jaclyn Ellis; Jin Li; Jeremy Walston; Leslie A Lange; Mary Cushman; Russell P Tracy Journal: Arterioscler Thromb Vasc Biol Date: 2012-11-15 Impact factor: 8.311