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目的研究结核分枝杆菌eis基因表达对人单核巨噬细胞Thp-1细胞的影响,筛选出差异性表达细胞因子,探讨结核分枝杆菌eis基因在单核巨噬细胞致持留的免疫机制。方法 eis基因重组耻垢分枝杆菌MS-pmv261-eis及对照组细菌MSpmv261感染经PMA诱导分化的人单核巨噬细胞Thp-1细胞;细胞信号传导通路抑制剂用于分析作用机制;ELISA方法检测细胞上清中IL-4、IL-6、IL-10、IL-12、INF-γ表达水平;筛选差异性表达的细胞因子,RT-PCR进一步确定该细胞因子的基因表达。结果结核分枝杆菌eis基因可引起IL-10表达增加(P<0.05),而对细胞因子IL-4、IL-6、IL-12、INF-γ的表达无显著影响(P>0.05),RT-PCR结果也进一步确证了IL-10基因的高表达。2-AP和U0126能明显抑制MS-pmv261-eis(MS-eis)感染引起的IL-10的释放(P<0.05)。结论结核分枝杆菌eis基因可上调Thp-1细胞IL-10在蛋白水平和基因水平的表达,可能通过MEK1/2或PKR信号通路刺激IL-10的释放来增强机体的免疫功能,其作用可能与结核分枝杆菌持留性有关。
Objective To study the effect of Mycobacterium tuberculosis eis gene expression on human monocyte-macrophage Thp-1 cells and to screen out differentially expressed cytokines to explore the immune mechanism of Mycobacterium tuberculosis eis gene in monocyte-macrophage-induced persistence. Methods eis gene recombinant Mycobacterium smegmatis MS-pmv261-eis and control group MSpmv261 were infected by PMA induced differentiation of human monocyte-macrophage Thp-1 cells; cell signaling pathway inhibitors for the analysis of the mechanism of action; ELISA method The expression of IL-4, IL-6, IL-10, IL-12 and INF-γ in the supernatant of the cells were detected. The differentially expressed cytokines were screened and the gene expression of the cytokines was further confirmed by RT-PCR. Results The eis gene of M. tuberculosis could induce the increase of IL-10 expression (P <0.05), but had no significant effect on the expression of IL-4, IL-6, IL-12 and INF- RT-PCR results further confirmed the high expression of IL-10 gene. 2-AP and U0126 significantly inhibited IL-10 release induced by MS-pmv261-eis (MS-eis) infection (P <0.05). Conclusion Mycobacterium tuberculosis eis gene can up-regulate the expression of IL-10 in Thp-1 cells at the protein and gene level, and may enhance the immune function by stimulating the release of IL-10 through MEK1 / 2 or PKR signaling pathway. And Mycobacterium tuberculosis retention related.