【摘要】 目的利用生物信息学技术探究1/17型辅助T(Th1/17)细胞的功能及其分化过程。方法采用高通量基因表达数据库(GEO)中包含来源于健康人体Th17细胞和Th1/17细胞基因表达数据的基因芯片数据集(GSE104021)进行生物信息学分析。以Th17细胞为对照,采用R语言软件分析Th17细胞和Th1/17细胞之间的差异表达基因,以此探究Th1/17细胞发挥功能的主要效应分子。后通过R语言软件对差异表达基因进行基因本体(GO)分析和京都基因与基因组百科全书(KEGG)分析。最后选用GO分析中被富集到目标生物过程中的基因,进行蛋白质相互作用网络(PPI)分析,探索Th1/17细胞的分化过程。结果差异表达基因分析结果显示,与Th17细胞相比, Th1/17细胞中低表达基因为白细胞介素17A(IL-17A)和CC趋化因子受体4(CCR4);高表达基因为含卷曲螺旋域3(CCDC3)、 CC趋化因子配体4(CCL4)、集落刺激因子2受体β亚单位(CSF2RB)、 CCL5、γ干扰素(IFNG)和上皮基质相互作用分子1(EPSTI1)。GO分析中细胞组分分析结果显示,这些差异基因表达产物主要定位于细胞膜胞外侧和粒细胞-巨噬细胞集落刺激因子(GM-CSF)受体复合物中;生物过程分析结果显示差异基因表达产物参与上调白细胞介素23(IL-23)产生,趋化因子介导的信号通路和上调自然杀伤(NK)细胞趋化性;分子功能分析结果显示差异表达基因表达产物具有CC趋化因子受体5(CCR5)结合活性,细胞因子活性和γ干扰素(IFN-γ)受体结合活性。KEGG分析结果显示差异基因被富集到细胞因子-细胞因子受体相互作用,类风湿性关节炎,趋化因子信号通路,炎症性肠病等通路中。GO分析结果显示,差异基因IL-17A和IFNG被富集到能够上调IL-23产生的生物过程中。PPI结果显示:IL-17A和IFNG具有调节细胞因子产生和调节髓样白细胞分化的生物功能。结论生物信息学分析结果显示Th1/17细胞高表达基因CCL4、 CSF2RB、 CCL5、 IFNG和EPSTI1编码的蛋白产物为Th1/17细胞潜在的功能效应分子。Th1/17产生IFN-γ和IL-17A,作用于来源于髓样白细胞的巨噬细胞和树突状细胞(DC),促进巨噬细胞和DC分化并产生IL-23。IL-23作用于Th17细胞,使其转分化为Th1/17细胞。更多还原

【Abstract】 Objective To investigate the function and differentiation of 1/17 type helper T(Th1/17) cells. MethodsBioinformatics analysis was performed using a gene chip dataset(GSE104021) in GEO which contains gene expression data from Th17 cells and Th1/17 cells of healthy human subjects. Taking Th17 cells as the control, R language software was used to analyze the differentially expressed genes(DEGs) between Th17 cells and Th1/17 cells, so as to explore the main functional molecules of Th1/17 cells. After that, gene ontology(GO) analysis and Kyoto encyclopedia of genes and genomes(KEGG) analysis of DEGs were conducted by R language software. Finally, the genes enriched into the target biological process in the GO analysis were selected for protein-protein interaction network(PPI) analysis to explore the differentiation process of Th1/17 cells. Results Analysis of DEGs showed that, compared with Th17 cells, the underexpressed genes in Th1/17 cells were interleukin 17A(IL-17A) and C-C motif chemokine receptor 4(CCR4). The over-expressed genes were coiled-coil domain-containing 3(CCDC3), C-C motif chemokine ligand 4(CCL4), colony stimulating factor 2 receptor beta common subunit(CSF2RB), C-C motif chemokine ligand 5(CCL5), interferon gamma(IFNG) and epithelial stromal interaction 1(EPSTI1). In GO analysis, cell component analysis showed that the expression products of these DEGs were mainly located at external side of plasma membrane and the granulocyte-macrophage colony stimulating factor(GM-CSF) receptor complex; biological process analysis showed that the expression products of DEGs were involved in the upregulation of interleukin 23(IL-23), the chemokine-mediated signaling pathway and the upregulated chemotaxis of natural killer(NK) cells; molecular function analysis showed that the expression products of these DEGs had C-C motif chemokine 5 receptor(CCR5) binding activity, cytokine activity and interferon gamma(IFN-γ) receptor binding activity. The results of KEGG analysis showed that the DEGs were enriched in the cytokine-cytokine receptor interaction, rheumatoid arthritis, inflammatory bowel disease and chemokine signaling pathways. The GO analysis showed that DEGs IL-17A and IFNG were enriched to the biological process of upregulating IL-23 production. PPI showed that IL-17A and IFNG had biological functions of regulating cytokine production and myeloid white blood cell differentiation. Conclusion Bioinformatics analysis showed that the protein products encoded by overexpressed genes CCL4, CSF2RB, CCL5, IFNG and EPSTI1 in Th1/17 cells were potential functional effectors of Th1/17 cells. Th1/17 cells could produce IFN-γ and IL-17A, which act on macrophages and dendritic cells(DCs) derived from myeloid white blood cells, thus promoting the differentiation of macrophages and DCs and the production of IL-23. IL-23 promotes trans-differentiation of Th17 cells into Th1/17 cells.更多还原