CD14-159C/T基因多态性对全血培养CD14表达的影响

目的探讨CD14基因启动子-159C/T基因多态性对全血培养CD14mRNA表达及可溶性CD14(sCD14)浓度的影响。方法采集118例健康献血员血标本,用全血细胞培养模型检测内毒素刺激前后CD14mRNA表达及sCD14浓度的变化。采用聚合酶链反应(PCR)及限制性内切酶HaeⅢ对PCR产物的消化作用检测CD14基因多态性。同时,对内毒素刺激后肿瘤坏死因子α(TNF-α)诱生水平进行了分析。结果118例健康献血员中,等位基因T和C的频率分别为60.2%和39.8%;40例是T等位基因纯合子(TT),62例为杂合子(TC),还有16例基因型为CC。基因型TT与TC白细胞中CD14mRNA的表达及上清液sCD14浓度均明显高于CC纯合子(P<0.05或0.01)。并且TT纯合子TNF-α诱生水平为(352±215)pg/ml,显著高于基因型TC及CC(261±163)pg/ml及(198±122)pg/ml,P<0.05]。结论内毒素受体CD14-159C/T基因多态性对全血培养CD14的表达及释放产生显著影响,并与内毒素诱导TNF-α的反应性相关。

CD14 genomic polymorphism influences CD14 expression in whole blood culture

OBJECTIVE: To investigate whether -159C/T promoter polymorphism of the CD14 gene influences the CD14 expression as well as release in whole blood culture. METHODS: One hundred and eighteen healthy human blood donors were included in the present study. CD14 mRNA expression and soluble CD14 levels were measured using a whole blood cell culture model with or without lipopolysaccharide (LPS) stimulation. The CD14 gene polymorphism was determined by polymerase chain reaction and subsequent HaeIII restriction enzyme digestion of the polymerase chain reaction products. TNF-alpha production in the whole blood culture was also measured. RESULTS: Among the 118 individuals, there were 40 subjects homozygous for the T allele (TT), 62 were heterozygous (CT), and 16 had the genotype CC. The CD14 mRNA expression in leukocytes and soluble CD14 levels in supernatant were higher in TT homozygotes and carriers of the genotype TC compared with individuals homozygous for the C allele (P < 0.05 or 0.01). In addition, the highest values of TNF-alpha production were found in TT homozygote (352 +/- 215) pg/ml] compared with both genotype TC and CC (261 +/- 163) pg/ml, (198 +/- 122) pg/ml, both P < 0.05]. CONCLUSION: The -159C/T promoter polymorphism of the LPS receptor CD14 may influence the CD14 expression as well as release in whole blood culture, and it might be associated with TNF-alpha response to LPS stimulation.