ESAT-6刺激结核患者CD4+ T淋巴细胞前后释放IFN-γmRNA差异表达及信号通路变化
张焰;王晓玮;程筱雯;徐元宏
【期刊名称】《安徽医科大学学报》 【年(卷),期】2016(051)012
【摘要】目的：利用全基因组表达谱芯片对早期分泌靶向抗原6(ESAT-6)刺激结核患者外周血CD4+ T淋巴细胞前后释放γ-干扰素( IFN-γ) mRNA进行检测,并对mRNA在刺激过程中可能涉及到的信号通路变化进行分析。方法抽取3例初诊结核患者外周静脉血各10 ml,密度梯度离心法分离获得外周血单个核细胞( PBMC), ESAT-6刺激PBMC,6 h后磁珠法分选CD4+ T淋巴细胞,提取细胞总RNA,全基因组表达谱芯片检测刺激前后差异表达基因。利用GO分析对差异表达的基因进行功能分类, KEGG 进行信号通路分析。结果 ESAT-6刺激后差异表达的mRNA共有2297条,相对高表达的 mRNA 有1071条,相对低表达的 mRNA 有1226条(>2倍变化且P<0．05)。信号通路分析显示差异表达的mRNA参与了17条信号通路的调控,其中JAK-STAT信号通路、Toll样受体信号通路、NF-κB信号通路、MAPK信号通路、PI3K-Akt信号通路等5条信号通路与结核释放IFN-γ关系密切。结论 ESAT-6刺激后的CD4+ T淋巴细胞有大量mRNA发生差异表达,其在多个与结核释放IFN-γ密切相关的信号通路中发挥着重要的调控作用。%Objective To detect the difference in release IFN-γ mRNA expression profiling of early secreted anti-genic target 6 ku protein(ESAT-6) stimulated CD4 + T lymphocytes from tuberculosis patients peripheral blood and also to identify the changes in signaling
pathways through whole genome expression microarray screening. Methods 10 ml peripheral venous blood was collected from three first diagnosed tuberculosis patients. Peripheral blood mononuclear cell( PBMC) was isolated by density gradient centrifugation and stimulated with ESAT-6 for 6 hours. Then, CD4 + T lymphocytes were obtained from ESAT-6 stimulated PBMC by immunomagnetic technique. Total RNA was extracted, useing whole genome expression microarray to detect differences in gene expression before and after stimulation. GO and KEGG were used to analyze the functional classification of differentially expressed genes and signaling pathways. Results After ESAT-6 stimulation, it had 2 297 differentially expressed mRNAs. 1 071 mRNAs were up-regulated, 1 226 mRNAs were down-regulated ( >2 fold changes and P <0. 05 ) . 17 signaling pathways were involved based on mRNAs expression data and 5 of which were involved in tuberculosis IFN-γ re-lease( JAK-STAT signaling pathway, toll like receptor signaling pathway, NF-κB signaling pathway, MAPK signa-ling pathway, and PI3K-Akt signaling pathway). Conclusion After ESAT-6 stimulation, obvious changes of mR-NAs expression profiling are observed. Those differentially expressed mRNAs play important roles in regulating sig-naling pathways in CD4 + T lymphocytes tuberculosis IFN-γ release. 【总页数】6页(1727-1732)
【关键词】早期分泌靶向抗原6;γ-干扰素;基因芯片;信号通路;结核
【作者】张焰;王晓玮;程筱雯;徐元宏
【作者单位】安徽医科大学第一附属医院检验科，合肥 230022;安徽医科大学第一附属医院检验科，合肥 230022;安徽医科大学第一附属医院检验科，合肥 230022;安徽医科大学第一附属医院检验科，合肥 230022 【正文语种】中文
【中图分类】R349.81;R52 【文献来源】
https://www.zhangqiaokeyan.com/academic-journal-cn_acta-universitatis-medicinalis-anhui_thesis/0201243313296.html 【相关文献】
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ESAT-6刺激结核患者CD4+ T淋巴细胞前后释放IFN-γmRNA差异表达及信号通路变化
