Immune-mediated inflammation must be down-regulated to facilitate tissue remodeling during homeostatic restoration of an inflammatory response. Uncontrolled or over-exuberant immune activation can cause autoimmune diseases, as well as tissue destruction. A151, the archetypal example of a chemically synthesized suppressive oligodeoxynucleotide (ODN) based on repetitive telomere-derived TTAGGG sequences, was shown to successfully down-regulate a variety of immune responses. However, the degree, duration and breadth of A151-induced transcriptome alterations remain elusive. Here, we performed a comprehensive microarray analysis in combination with Ingenuity Pathway Analysis (IPA) using murine splenocytes to investigate the underlying mechanism of A151-dependent immune suppression. Our results revealed that A151 significantly down-regulates critical mammalian target of rapamycin (mTOR) activators (Pi3kcd, Pdpk1 and Rheb), elements downstream of mTOR signaling (Rps6ka1, Myc, Stat3 and Slc2a1), an important component of the mTORC2 protein complex (Rictor) and Mtor itself. The effects of A151 on mTOR signaling were dose- and time-dependent. Moreover, flow cytometry and immunoblotting analyses demonstrated that A151 is able to reverse mTOR phosphorylation comparably to the well-known mTOR inhibitor rapamycin. Furthermore, Seahorse metabolic assays showed an A151 ODN-induced decrease in both oxygen consumption and glycolysis implying that a metabolically inert state in macrophages could be triggered by A151 treatment. Overall, our findings suggested novel insights into the mechanism by which the immune system is metabolically modulated by A151 ODN.

中文翻译：

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表达抑制性寡核苷酸的TTAGGG主题通过mTOR信号传导途径调节细胞能量。

免疫介导的炎症必须下调，以促进炎症反应的稳态恢复过程中的组织重塑。不受控制或过度旺盛的免疫激活会导致自身免疫性疾病以及组织破坏。A151是基于重复端粒衍生的TTAGGG序列的化学合成的抑制性寡脱氧核苷酸（ODN）的原型示例，已显示成功下调了多种免疫应答。然而，A151诱导的转录组改变的程度，持续时间和广度仍然难以捉摸。在这里，我们进行了综合的微阵列分析，结合使用鼠脾细胞的机能途径分析（IPA），研究了A151依赖性免疫抑制的潜在机制。我们的研究结果表明，A151显着下调了雷帕霉素（mTOR）激活剂（Pi3kcd，Pdpk1和Rheb）的关键哺乳动物靶标，mTOR信号下游的元素（Rps6ka1，Myc，Stat3和Slc2a1），这是mTORC2蛋白复合物的重要组成部分（ Rictor）和Mtor本身。A151对mTOR信号传导的影响是剂量和时间依赖性的。此外，流式细胞仪和免疫印迹分析表明，A151与已知的mTOR抑制剂雷帕霉素相比，能够逆转mTOR的磷酸化。此外，海马代谢测定法显示，A151 ODN诱导的耗氧量和糖酵解均减少，这暗示巨噬细胞中的代谢惰性状态可以通过A151治疗触发。总体，