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Identification of genes differentially expressed in T cells following stimulation with the chemokines CXCL12 and CXCL10

机译:鉴定趋化因子CXCL12和CXCL10刺激后T细胞中差异表达的基因

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Background Chemokines are involved in many biological activities ranging from leukocyte differentiation to neuronal morphogenesis. Despite numerous reports describing chemokine function, little is known about the molecular changes induced by cytokines. Methods We have isolated and identified by differential display analysis 182 differentially expressed cDNAs from CXCR3-transfected Jurkat T cells following treatment with CXCL12 or CXCL10. These chemokine-modulated genes were further verified using quantitative RT-PCR and Western blot analysis. Results One hundred and forty-six of the cDNAs were successfully cloned, sequenced, and identified by BLAST. Following removal of redundant and non-informative clones, seventeen mRNAs were found to be differentially expressed post treatment with either chemokine ligand with several representing known genes with established functions. Twenty-one genes were upregulated in these transfected Jurkat cells following both CXCL12 and CXCL10, four genes displayed a discordant response and seven genes were downregulated upon treatment with either chemokine. Identified genes include geminin (GEM), thioredoxin (TXN), DEAD/H box polypeptide 1 (DDX1), growth hormone inducible transmembrane protein (GHITM), and transcription elongation regulator 1 (TCERG1). Subsequent analysis of several of these genes using semi-quantitative PCR and western blot analysis confirmed their differential expression post ligand treatment. Conclusions Together, these results provide insight into chemokine-induced gene activation and identify potentially novel functions for known genes in chemokine biology.
机译:背景趋化因子参与许多生物学活动,从白细胞分化到神经元形态发生。尽管有许多描述趋化因子功能的报道,但关于细胞因子诱导的分子变化知之甚少。方法我们通过差异显示分析从CXCL12或CXCL10处理后,从CXCR3转染的Jurkat T细胞中分离并鉴定了182个差异表达的cDNA。这些趋化因子调节的基因使用定量RT-PCR和Western blot分析进一步验证。结果通过BLAST成功地克隆了146个cDNA。去除冗余和非信息性克隆后,发现用任一趋化因子配体处理后有十七种mRNA差异表达,其中几种趋化因子配体具有已知的已确定功能的基因。在使用CXCL12和CXCL10进行转染后,这些转染的Jurkat细胞中有21个基因上调,其中四个基因表现出不一致的反应,而七个基因在用任一趋化因子处理后均下调。鉴定的基因包括geminin(GEM),thioredoxin(TXN),DEAD / H box多肽1(DDX1),生长激素诱导型跨膜蛋白(GHITM)和转录延伸调控因子1(TCERG1)。随后使用半定量PCR和蛋白质印迹分析对其中一些基因进行了分析,证实了配体处理后它们的差异表达。结论在一起,这些结果提供了对趋化因子诱导的基因激活的了解,并鉴定了趋化因子生物学中已知基因的潜在新功能。

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