结核病患者免疫相关基因表达变化的基因芯片研究

Study of the expression profile of immunogenesis associated genes in tuberculosis by microarray

目的探讨免疫相关基因在结核病患者体内的差异表达。方法应用基因芯片技术和生物信息学方法,对63例单纯结核性胸膜炎患者胸腔积液及其外周血中单核细胞与69例肺癌患者的癌性胸腔积液和其外周血中单核细胞的mRNA表达水平进行分析比较,所做的表达谱杂交检测均重复1次并在荧光交换后再重复1次以抑制噪声,对阳性和部分阴性结果进行northernblotting复验。结果在对基因表达谱原始数据进行标准化校正、聚类分析和统计学处理后,发现在626条免疫相关基因中差异表达基因53条,其中与细胞增殖相关的基因16个,与细胞免疫应答相关的基因14个,与信号传导有关的基因20个,其他相关基因3个;在结核病患者胸腔积液淋巴细胞中tnfΑ、iglλ、il-17、il-17r、hladp、lcp-1、tcr-Α、tcr-Β、hsp-75、cxcr4、fyb、hlag、hlaa、il18bp、il2r、ltΒ、il-8、ip-10、mcp-1、il-12、il-12r、il-10、canx、irf2、ifn-Γ、tlr、il-1、il-7、tlr、lsp-1、il1-4共31个基因表达较对照组增高4倍以上,il-4、il-18、il-15、ifg-1、scya-14、ablim、peci、ppid、hsf-2、act-2、maoa、ttid、gatm、tgfb3、insr、thb-d、trap-1、tcr-Γ、tcrΔ、il1-3r、il1-1、igf-1、a2m共22个基因表达水平较对照组降低4倍以上。结论机体抗结核免疫的发生、发?

Objective To explore the difference of gene expression profile in tuberculosis patients. Methods mRNA levels of pleural fluid and peripheral blood mononuclear cells(PBMC) in tuberculous pleurisy and lung cancer patients were compared by cDNA microarray. Paired mRNAs from fluid specimens of tuberculosis and lung cancer cases were labeled with different fluorochromes during cDNA probe synthesis in a reverse-transcription reaction. The signal intensity of each spot was measured by laser scanner and gene expression was quantified as the tubercle-to-normal fluorescence ratio (T∶N ratio). The gene was defined as over expression when the T∶N ratio was greater than 2.0 and under expression when the ratio was less than 0.5. Results Among 626 immunogenesis associated genes there were 53 differences, of which 31 (tnf-α,iglλ,il-17, il-17r, hla-dp, lcp1, tcrα, tcrβ, hsp75, cxcr4, fyb,hla-g, hla-a, il18bp, il-2r, lt-β, il-8, ip-10, mcp-1, il-12, il-12r, il-10, canx, irf2, ifn-γ, tlr, il-1, il-7, tlr, lsp-1, il-14)were higher and 22 (il-4, il-18, il-15, ifg-1, scya14, ablim, peci, ppid, hsf 2, actg2, maoa, ttid, gatm, tgfb3, insr, thbd,trap1, tcrγ, tcrδ, il-13r, il-11, igf1, a2m)were lower in tuberculous pleurisy than those in the control. Conclusions The immunogenesis of tuberculosis involves multi-genetic expression changes, such as tnf-α, il-17, il-12, tcrα,tcrβ,hsp75,cxcr4,il-4,il-18,il-15 etc., the expression profile of which changed dramatically. The results provide new insight for understanding of the pathogenic mechanisms of tuberculosis and exploring new therapeutic strategies.