抑制g - αs/cAMP信号可降低tcr刺激的CD4(+) T辅助细胞IL-2转录。

Q2 Biochemistry, Genetics and Molecular Biology Journal of Molecular Signaling Pub Date : 2015-07-06 DOI:10.5334/1750-2187-10-2

Thomas R Hynes, Evan A Yost, Stacy M Yost, Cassandra M Hartle, Braden J Ott, Catherine H Berlot

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引用次数: 5

摘要

背景:cAMP在调节T细胞活化和功能中的作用一直存在争议。cAMP通常被认为是一种免疫抑制剂，但它也是产生最佳免疫反应所必需的。由于cAMP的作用可能取决于其细胞环境，本研究探讨了Gαs和腺苷酸环化酶的激活机制是否影响它们对T细胞受体(TCR)刺激的白介素-2 (IL-2) mRNA水平的影响。方法:比较阻断Gs偶联受体(GsPCR)介导的Gs活化对tcr刺激的CD4(+) T细胞IL-2 mRNA水平的影响与抑制Gαs表达或抑制腺苷酸环化酶活性的影响。比较了抑制Gαs表达对tcr刺激的cAMP积累的影响和阻断GsPCR信号的影响。结果:ZM-241385是一种gs偶联A2A腺苷受体(A2AR)拮抗剂，可提高tcr刺激的人CD4(+) T辅助细胞和Jurkat T细胞中IL-2 mRNA水平。g - α -s的显性负构建体g - α sdn - 3也增强了tcr刺激的IL-2 mRNA水平。与GsPCR拮抗剂类似，g - α sdn - 3阻断GsPCR依赖的g - αs和g - βγ的激活。相反，g - αs siRNA和腺苷酸环化酶抑制剂2'，5'-二脱氧腺苷(ddA)可降低tcr刺激的IL-2 mRNA水平。g - αs siRNA抑制tcr刺激下cAMP的合成，而g - α sdn 3则无此作用。ZM-241385增强IL-2 mRNA水平需要至少两天的TCR刺激，在TCR刺激三天后添加ddA可增强IL-2 mRNA水平。结论:GsPCRs在tcr刺激下IL-2 mRNA水平的调节中起抑制作用，而g - αs和cAMP则起刺激作用。此外，tcr依赖性的g - αs激活似乎不涉及gspcr。这些结果表明，Gαs/cAMP活化的环境和T细胞活化和分化的阶段决定了tcr刺激IL-2 mRNA水平的影响。

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Inhibition of Gαs/cAMP Signaling Decreases TCR-Stimulated IL-2 transcription in CD4(+) T Helper Cells.

Background: The role of cAMP in regulating T cell activation and function has been controversial. cAMP is generally known as an immunosuppressant, but it is also required for generating optimal immune responses. As the effect of cAMP is likely to depend on its cellular context, the current study investigated whether the mechanism of activation of Gαs and adenylyl cyclase influences their effect on T cell receptor (TCR)-stimulated interleukin-2 (IL-2) mRNA levels.

Methods: The effect of blocking Gs-coupled receptor (GsPCR)-mediated Gs activation on TCR-stimulated IL-2 mRNA levels in CD4(+) T cells was compared with that of knocking down Gαs expression or inhibiting adenylyl cyclase activity. The effect of knocking down Gαs expression on TCR-stimulated cAMP accumulation was compared with that of blocking GsPCR signaling.

Results: ZM-241385, an antagonist to the Gs-coupled A2A adenosine receptor (A2AR), enhanced TCR-stimulated IL-2 mRNA levels in primary human CD4(+) T helper cells and in Jurkat T cells. A dominant negative Gαs construct, GαsDN3, also enhanced TCR-stimulated IL-2 mRNA levels. Similar to GsPCR antagonists, GαsDN3 blocked GsPCR-dependent activation of both Gαs and Gβγ. In contrast, Gαs siRNA and 2',5'-dideoxyadenosine (ddA), an adenylyl cyclase inhibitor, decreased TCR-stimulated IL-2 mRNA levels. Gαs siRNA, but not GαsDN3, decreased TCR-stimulated cAMP synthesis. Potentiation of IL-2 mRNA levels by ZM-241385 required at least two days of TCR stimulation, and addition of ddA after three days of TCR stimulation enhanced IL-2 mRNA levels.

Conclusions: GsPCRs play an inhibitory role in the regulation of TCR-stimulated IL-2 mRNA levels whereas Gαs and cAMP can play a stimulatory one. Additionally, TCR-dependent activation of Gαs does not appear to involve GsPCRs. These results suggest that the context of Gαs/cAMP activation and the stage of T cell activation and differentiation determine the effect on TCR-stimulated IL-2 mRNA levels.

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Journal of Molecular Signaling Biochemistry, Genetics and Molecular Biology-Biochemistry

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期刊介绍： Journal of Molecular Signaling is an open access, peer-reviewed online journal that encompasses all aspects of molecular signaling. Molecular signaling is an exponentially growing field that encompasses different molecular aspects of cell signaling underlying normal and pathological conditions. Specifically, the research area of the journal is on the normal or aberrant molecular mechanisms involving receptors, G-proteins, kinases, phosphatases, and transcription factors in regulating cell proliferation, differentiation, apoptosis, and oncogenesis in mammalian cells. This area also covers the genetic and epigenetic changes that modulate the signaling properties of cells and the resultant physiological conditions.