cAMP差异调控CD39^(+)、CTLA-4^(+)、PD-1^(+)T细胞亚群产生  

cAMP differentially regulates production of CD39^(+),CTLA-4^(+)and PD-1^(+)T cell subsets

作　　者：蔡雨俊 郝振平 郑永唐[2] 田仁荣[2] CAI Yujun;HAO Zhenping;ZHENG Yongtang;TIAN Renrong(College of Pharmacy,Dali University,Dali 671000,China;Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences,Kunming Institute of Zoology,Chinese Academy of Science,Kunming 650223,China;School of Life Sciences,University of Science and Technology of China,Hefei 230026,China)

机构地区：[1]大理大学药学院,大理671000 [2]中国科学院昆明动物研究所,中国科学院动物模型与人类疾病机理重点实验室,昆明650223 [3]中国科学技术大学生命科学学院,合肥230026

出　　处：《中国免疫学杂志》2024年第12期2471-2477,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金(32070181);云南省基础研究计划(2019FA041)。

摘　　要：目的:分析CD39、CTLA-4及PD-1在T细胞上的表达及表达模型,探讨环磷酸腺苷(cAMP)对其表达的影响,解析调控其表达的关键通路。方法:通过腺苷酸环化酶(ACs)激活剂、磷酸二酯酶(PDE)抑制剂、蛋白激酶A(PKA)抑制剂以及PKA-CREB抑制剂等小分子化合物体外刺激猕猴外周血单个核细胞,流式细胞术检测CD39、CTLA-4、PD-1阳性(CD39^(+)、CTLA-4^(+)、PD-1^(+))T细胞的比率变化,分析其表达模式,对比小分子对表达/共表达的阳性细胞比率的影响,明确表达模式及调控表达的分子与通路。结果:正常猕猴的CD4^(+)T和CD8^(+)T细胞中CD39^(+)、CTLA-4^(+)和PD-1^(+)细胞比率较低,且阳性细胞主要表达其中一种分子;增加胞内cAMP浓度,不影响CD39^(+)T细胞比率,显著增加CTLA-4^(+)T细胞比率,降低PD-1^(+)CD8^(+)T细胞比率,涉及细胞群为CTLA-4和PD-1单表达的细胞群;抑制PKA活性可降低PDE广谱抑制剂3-异丁基-1-甲基黄嘌呤(IBMX)对CTLA-4表达的增效作用,但不影响PD-1的表达;交换蛋白激动剂不影响CTLA-4的表达,但下调PD-1^(+)CD4^(+)/CD8^(+)T细胞比率;PDE4B选择性抑制剂对CTLA-4的上调作用与IBMX相似,而PDE3和PDE5A选择性抑制剂对PD-1表达的调控与IBMX相似。结论:CD39、CTLA-4和PD-1在T细胞上有不同表达模型,但受cAMP水平不同的调控,且参与表达调控的cAMP下游信号通路不同。Objective:To analyze expression and models of CD39,CTLA-4,and PD-1 on T cells,to investigate effect of cyclic adenosine monophosphate(cAMP)on their expressions,and to analyze key pathways regulating their expressions.Methods:Small molecules such as adenylate cyclase(ACs)activators,phosphodiesterase(PDE)inhibitors,protein kinase A(PKA)inhibitors and PKA-CREB inhibitors were used to stimulate rhesus macaque peripheral blood mononuclear cells in vitro,and changes in ratios of CD39,CTLA-4,and PD-1 positive(CD39^(+),CTLA-4^(+),and PD-1^(+))T cells were detected by flow cytometry,and to analyze their expression patterns,and effects of small molecules on ratio of positively expressing/co-expressing cells were compared to clarify expression patterns and molecules and pathways that regulated expression.Results:Normal macaque CD4^(+)T and CD8^(+)T cells had low ratios of CD39^(+),CTLA-4^(+),and PD-1^(+)cells,and positive cells predominantly expressed only one of these molecules.Increasing intracellular cAMP levels did not affect CD39^(+)T cell ratio,significantly increased CTLA-4^(+)T cell ratio,and decreased PD-1^(+)CD8^(+)T cell ratio,involving cell populations with mono expression of CTLA-4 and PD-1.Inhibition of PKA activity reduced potentiation of CTLA-4 expression by broad-spectrum PDE inhibitor,3-isobutyl-1-methylxanthine(IBMX),but did not affect PD-1 expression.Exchange protein activated by cAMP did not affect CTLA-4 expression but downregulate PD-1^(+)CD4^(+)/CD8^(+)T cells ratio.Upregulation of CTLA-4 by PDE4B selective inhibitors was similar to IBMX,while regulation of PDE3 and PDE5A selective inhibitors on PD-1 expression was similar to IBMX.Conclusion:CD39,CTLA-4 and PD-1 have different expression models on T cells but are differentially regulated by cAMP levels and have different cAMP downstream signal pathways involved in expression regulation.

关 键 词：CD39 CTLA-4 PD-1 CAMP T细胞 

分 类 号：R392[医药卫生—免疫学]