CD14-c159T基因多态性对哮喘儿童外周血单个核细胞内毒素细胞因子反应的影响。

The effect of CD14-c159T genotypes on the cytokine response to endotoxin by peripheral blood mononuclear cells from asthmatic children.

作者信息

Keskin Ozlem, Birben Esra, Saçkesen Cansin, Soyer Ozge U, Alyamaç Evrim, Karaaslan Cağatay, Tokol Nihan, Ercan Hülya, Kalayci Omer

机构信息

Pediatric Allergy and Asthma Unit, Hacettepe University School of Medicine, Ankara, Turkey.

出版信息

Ann Allergy Asthma Immunol. 2006 Sep;97(3):321-8. doi: 10.1016/S1081-1206(10)60796-X.

DOI:10.1016/S1081-1206(10)60796-X

PMID:17042137

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).

CONCLUSION

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.

摘要

背景

CD14启动子第159位的C-T多态性（C-159T）可调节细胞对内毒素的反应，并显著影响总IgE水平。这种基因变异对炎症细胞细胞因子反应的影响尚未完全明确。

目的

研究CD14-C159T基因型对哮喘儿童外周血单个核细胞（PBMC）对内毒素反应的影响。

方法

将CD14启动子TT基因型（n = 11）和CC基因型（n = 11）的哮喘儿童的PBMC在100 ng/mL内毒素、10 μg/mL伴刀豆球蛋白A或仅培养基存在的情况下进行培养。通过酶联免疫吸附测定法测定培养上清液中可溶性CD14（sCD14）、白细胞介素（IL）-1β、IL-4、IL-10、IL-12、IL-13、干扰素-γ和转化生长因子β的浓度，并使用逆转录聚合酶链反应评估转录差异。

结果

在未刺激条件下，与CC基因型儿童相比，TT基因型儿童向培养上清液中分泌的sCD14水平更高（P = 0.03，Mann-Whitney U检验）。内毒素刺激后，TT基因型儿童培养上清液中的IL-10和IL-1β浓度均显著更高（分别通过协方差分析[ANCOVA]，P = 0.02和P = 0.009）。IL-10和sCD14的信使核糖核酸表达与蛋白质浓度结果一致。伴刀豆球蛋白A刺激导致TT基因型儿童的IL-4水平较低（P = 0.02，ANCOVA）。