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

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P2 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.