title: The effect of CD14-c159T genotypes on the cytokine response to endotoxin by peripheral blood mononuclear cells from asthmatic children.
abstract: BACKGROUND
  A C-T polymorphism at position 159 in the promoter of CD14 (C-159T) modulates the cellular response to endotoxin and significantly influences total IgE levels. The effect of this genetic variant on the cytokine response of the inflammatory cells is incompletely understood.
  
  
  OBJECTIVE
  To investigate the effects of CD14-C159T genotypes on the response to endotoxin by peripheral blood mononuclear cells (PBMCs) in children with asthma.
  
  
  METHODS
  The PBMCs from asthmatic children with the TT (n = 11) and CC (n = 11) genotypes at the CD14 promoter were cultured in the presence of endotoxin, 100 ng/mL; concanavalin A, 10 microg/mL; or medium alone. Concentrations of soluble CD14 (sCD14), interleukin (IL) 1beta, IL-4, IL-10, IL-12, IL-13, interferon-gamma, and transforming growth factor beta were determined in culture supernatants by enzyme-linked immunosorbent assay, and the transcriptional differences were evaluated using reverse-transcriptase polymerase chain reaction.
  
  
  RESULTS
  Under unstimulated conditions, children with the TT genotype produced higher levels of sCD14 into the culture supernatant compared with children with the CC genotype (P = .03, Mann Whitney U test). Both IL-10 and IL-1beta concentrations were significantly higher in culture supernatants of children with the TT genotype after endotoxin stimulation (P = .02 and P = .009, respectively, by analysis of covariance [ANCOVA]). Messenger RNA expression was consistent with the results of protein concentration for IL-10 and sCD14. Concanavalin A stimulation resulted in lower levels of IL-4 in children with the TT genotype (P = .02, ANCOVA).
  
  
  CONCLUSIONS
  The genotype at the CD14 promoter C159T locus may significantly influence the cytokine response of PBMCs obtained from asthmatic children. Differences in IL-10 and IL-4 production by alternative genotypes may contribute to the observed genotype effect on total IgE.
authors.affiliation: Pediatric Allergy and Asthma Unit, Hacettepe University School of Medicine, Ankara, Turkey.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Keskin
authors.forename: Esra
authors.initials: E
authors.lastname: Birben
authors.forename: Cansin
authors.initials: C
authors.lastname: Saçkesen
authors.forename: Ozge U
authors.initials: OU
authors.lastname: Soyer
authors.forename: Evrim
authors.initials: E
authors.lastname: Alyamaç
authors.forename: Cağatay
authors.initials: C
authors.lastname: Karaaslan
authors.forename: Nihan
authors.initials: N
authors.lastname: Tokol
authors.forename: Hülya
authors.initials: H
authors.lastname: Ercan
authors.forename: Omer
authors.initials: O
authors.lastname: Kalayci
mesh_terms: D000293:Adolescent; D001249:Asthma / Q000276:immunology*; D002478:Cells, Cultured; D002648:Child; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology*; D004731:Endotoxins / Q000276:immunology*; D005260:Female; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology* / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000235:genetics* / Q000276:immunology; D008297:Male; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D016207:Cytokines; D004731:Endotoxins; D018950:Lipopolysaccharide Receptors