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白细胞介素-6基因启动子单核苷酸多态性对其蛋白表达水平及结核易感性的影响

The influence of single nucleotide polymorphisms in the promoter region of IL-6 gene on its protein expression and the susceptibility to tuberculosis

摘要:

目的 鉴定位于IL-6基因启动子区影响结核易感性的单核苷酸多态性(SNP)位点,探讨SNP对IL-6分泌水平的影响.方法 采用病例-对照的研究方法,借助MassARRAY飞行时间质谱技术检测研究队列(495例结核患者和358例健康对照)IL-6基因6个SNP位点(rs13306436、rs2069824、rs 1800795、rs36215814、rs1800797、rs1800796)基因型,通过x2检验比较两组人群SNP位点等位基因频率差异,并计算各种等位基因型人群发生结核的风险性.应用免疫磁珠技术从结核患者PBMC中分选CD14+单核细胞,使用结核菌裂解产物(Mtb)、19×103脂蛋白进行刺激,通过ELISA检测不同基因型CD14+单核细胞上清中IL-6分泌水平的差异.结果 健康对照人群中6个SNP位点的基因型分布均符合Hardy-Weinberg遗传平衡检验.rs 1800796位点3种基因型CC、CG和GG在结核组和对照组的频率分别为64.5%、30.7%、4.8%和56.4%、34.9%、8.7%,结核组G等位基因频率(21.3%)显著低于对照组(26.1%),差异有统计学意义(P=0.02),携带C等位基因人群患结核风险明显升高(OR=1.40,95% CI=1.11 ~1.75).其余SNP位点等位基因频率分布在两组间差异无统计学意义.在19×103脂蛋白刺激下,rs 1800796位点GG基因型单核细胞上清IL-6浓度为(490.10±55.08) pg/ml,分泌水平显著低于CC或CG基因型(P<0.05),而使用Mtb刺激组间差异无统计学意义.结论 首次在汉族人群中发现IL-6基因rs1800796位点影响结核易感性,C为易感等位基因.rs1800796位点多态性能够影响IL-6蛋白表达水平,属于功能性SNP位点.

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abstracts:

Objective To explore the relationships between single nucleotide polymorphisms (SNP) in IL-6 gene promoter region and susceptibility to tuberculosis,and to investigate the influences of SNP on IL-6 production in various genotypes.Methods A case-control study was conducted on 495 patients with tuberculosis and 358 healthy controls.Sequenom MassARRAY system was used to detect genotypes of 6 SNPs (rs13306436,rs2069824,rs36215814,rs 1800797,rs1800796,and rs1800795) in IL-6 gene.Allelic frequency in cases and controls was compared using Chi-square test and odds ratio (OR) was calculated to clarify the association with risk for tuberculosis.CD14+ monocytes isolated from PBMCs of the subjects with MACS beads were stimulated with Mtb lysate and 19 × 103 lipoprotein.Then the concentrations ofIL-6 in supernatants of CD14+ monocytes with different genotypes were determined by ELISA.Results Genotype distribution of 6 SNPs in healthy controls was in accordance with Hardy-Weinberg equilibrium.G enotype frequencies of rs1800796 were 64.5% (CC),30.7% (CG),4.8% (GG) in TB group and 6.4%(CC),34.9% (CG),8.7% (GG) in control group,respectively.The allele frequency of G in TB group (21.3%) was lower than that in control group (26.1%) (P value =0.02),and allele C carriers were at higher risk for TB (OR 1.40,95% CI=1.11-1.75).There were no significant differences in allele frequencies for other SNPs.With 19×103 lipoprotein stimulation,the concentration of IL-6 in CD14+ monocytes isolated from individuals with rs1800796 GG genotype was significantly less than those with CC or CG genotype.However,there was no difference observed among different genotypes with Mycobacterium tuberculosis (Mtb)stimulation.Conclusion This is the first report showing that IL-6 gene rs1800796 polymorphism with C as the susceptible allele was associated with the susceptibility to tuberculosis in Han nationality.Moreover,as a functional SNP,rs1800796 polymorphism could affect IL-6 production.

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作者: 张国良 [1] 文玉欣 [2] 张明霞 [1] 詹森林 [1] 杨林 [2] 王正 [2] 陆坚 [1] 陈心春 [1]
栏目名称: 基础免疫学
DOI: 10.3760/cma.j.issn.0254-5101.2013.04.011
发布时间: 2013-06-19
基金项目:
国家自然科学基金 广东省自然科学基金 深圳市科技计划项目
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