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中国防痨杂志 ›› 2015, Vol. 37 ›› Issue (1): 35-39.doi: 10.3969/j.issn.1000-6621.2015.01.008

• 论著 • 上一篇    下一篇

不同毒性Mtb菌株对Ⅰ型干扰素基因的诱导作用及其在结核病患者外周血中的表达

詹森林 张国良 钟红剑 金晓菲 林巧 张明霞 陈心春 周伯平   

  1. 518112  广东医学院附属深圳市第三人民医院  广东省新发传染病诊治重点实验室（詹森林、张国良、金晓菲、张明霞、陈心春、周伯平）；深圳市宝安区慢性病防治院结核科（钟红剑、林巧）
  • 收稿日期:2014-10-28 出版日期:2015-01-10 发布日期:2015-02-08
  • 通信作者: 周伯平 E-mail:zhoubp@hotmail.com
  • 基金资助:

    国家自然科学基金（81172732，81341128）；广东省自然科学基金（S2012040007213）；深圳市科技计划项目（201202189）；深圳市科技计划基础研究项目（JCYJ20140411111718166）

The induction of type Ⅰ interferon by different M. tuberculosis strains and its expression in the peripheral blood of the patients with tuberculosis

ZHAN Sen-lin, ZHANG Guo-liang, ZHONG Hong-jian, JIN Xiao-fei, LIN Qiao, ZHANG Ming-xia, CHEN Xin-chun, ZHOU Bo-ping   

  1. Guangdong Key Laboratory for Emerging Infectious Disease, the Affiliated Shenzhen Third Hospital，Guangdong Medical College，Shenzhen 518112，China
  • Received:2014-10-28 Online:2015-01-10 Published:2015-02-08
  • Contact: ZHOU Bo-ping E-mail:zhoubp@hotmail.com

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摘要： 目的 研究不同Mtb菌株对Ⅰ型干扰素基因的诱导作用及其在结核病患者外周血中的表达。 方法 从2012年2月至2012年8月在深圳市第三人民医院健康体检者中利用随机数字表法抽取30名健康人外周血标本，并提取单个核细胞中分选出CD14+细胞，用重组人巨噬细胞集落刺激因子（macrophage colony stimulating factor，M-CSF）刺激，诱导分化为人巨噬细胞，并分别感染Mtb减毒株H37Ra和Mtb标准株H37Rv，应用实时定量PCR（real-time PCR）检测2种菌株感染后Ⅰ型干扰素基因IFNB和IFNA的相对表达量2-ΔΔCt值。此外使用上述方法对同期选取的结核病组（15例）、健康对照组（15例）、Mtb潜伏感染组（15例）的Ⅰ型干扰素基因的相对表达量进行检测。采用GraphPad 4.0软件对数据进行统计处理，两组均数间比较采用t检验，三组均数间比较采用方差分析，以P<0.05为差异有统计学意义。 结果 经H37Rv感染的健康人巨噬细胞的IFNB基因相对表达量2-ΔΔCt值（10.38±2.24）显著高于空白对照组（6.26±3.42），差异有统计学意义（t=2.47, P=0.026）；经H37Ra感染的巨噬细胞的IFNB基因相对表达量2-ΔΔCt值（8.92±0.85）与空白对照组（6.26±3.42）比较差异无统计学意义（t=1.84, P=0.09）；IFNA相对基因表达量2-ΔΔCt值在H37Rv感染组（5.11±2.31）、H37Ra感染组（5.17±3.40）及空白对照组（4.41±1.69）之间差异无统计学意义（F=0.16，P=0.85）。结核病组IFNB基因表达（4.32±1.22）显著高于健康对照组（2.73±1.23），差异有统计学意义（t=3.55, P=0.0014）；IFNA基因在健康对照组（3.91±0.75）、Mtb潜伏感染组（4.25±1.03）、结核病组（4.73±1.44）之间表达差异无统计学意义（F=1.93，P=0.064）。 结论 Mtb的感染可以诱导Ⅰ型干扰素基因的表达，在结核病患者中IFNB基因表达上调显著。

关键词: 结核, 干扰素Ⅰ型 , 基因表达

Abstract: Objective To study the induction of type I interferon (IFN) by different M. tuberculosis (Mtb) strains and its expression in the peripheral blood of the patients with tuberculosis.  Methods CD14+ cells were isolated from the peripheral blood mononuclear cells of 30 healthy people from Feb 2012 to Aug 2012, and were stimulated by the recombinant human macrophage colony stimulating factor (M-CFS) for 7 days, and differentiated into human macrophages. These macrophages were infected with Mtb strains H37Ra and H37Rv, respectively. The expression levels of type Ⅰ IFN genes IFNB and IFNA in the macrophages were detected with real-time PCR assay. We also observed the gene expression of type Ⅰ IFN in different population (15 healthy controls, 15 persons with latent Mtb infection and 15 patients with tuberculosis). Using GraphPad 4.0 software for statistical analysis, P<0.05 was considered statistically significant.  Results After Mtb H37Rv infection, IFNB gene expression in human macrophages was significantly higher than that in the control group (10.38±2.24 vs 6.26±3.42; t=2.47, P=0.026). There were no significant differences in IFNB gene expression between Mtb H37Ra infection group and control group (8.92±0.85 vs 6.26±3.42;t=1.84, P=0.09). There were no significant differences in IFNA gene expression among Mtb H37Rv infection group, H37Ra infection group and control group (5.11±2.31 vs 5.17±3.40 vs 4.41±1.69; F=0.16，P=0.85). The IFNB gene expressions in TB patients (4.32±1.22) were significantly higher than that in healthy controls (2.73±1.23; t=3.55, P=0.0014). There were no significant differences in IFNA gene expression among healthy controls, the persons with latent Mtb infection and the patients with tuberculosis (3.91±0.75 vs 4.25±1.03 vs 4.73±1.44；F=1.93，P=0.064).  Conclusion Virulent Mycobacterium tuberculosis infection can induce type Ⅰ IFN gene expression. Up-regulated expression of IFNB gene can be used as a potential molecular marker in the diagnosis of tuberculosis.

Key words: Tuberculosis, Interferon type Ⅰ, Gene expression

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