Quast Christine, Alter Christina, Ding Zhaoping, Borg Nadine, Schrader Jürgen
From the Division of Cardiology, Pulmonary Diseases and Vascular Medicine (C.Q.) and Department of Molecular Cardiology (C.Q., C.A., Z.D., N.B., J.S.), University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Germany.
Circ Heart Fail. 2017 Apr;10(4). doi: 10.1161/CIRCHEARTFAILURE.116.003346.
Structural damage during heart failure development leads to increased infiltration of leukocytes. Because purinergic signaling on immune cells may impact on the inflammatory response, we evaluated the role of ecto-5'-nucleotidase (CD73) on the development of heart failure after transverse aortic constriction (TAC) using global and T-cell-specific CD73 mice.
Leukocytes infiltrating the failing heart were analyzed by a multistep enzymatic procedure over a period of 16 weeks using fluorescence-activated cell sorting. TAC significantly enhanced the infiltration of leukocytes, especially T cells. The fraction of CD73 expressing cells increased over time exclusively on cytotoxic T cells, T-helper cells, and regulatory T cells. Cardiac function significantly declined in T-cell-specific CD4-CreCD73 mice identical to that observed in global CD73 mutants and was associated with enhanced fibrosis (collagen, laminin, vimentin, periostin). Expression analysis by quantitative reverse transcription polymerase chain reaction of extracellular purine degrading enzymes and P1 and P2 receptors on T cells isolated from the injured heart revealed profound upregulation of the enzymatic machinery for hydrolysis of extracellular adenosine triphosphate and nicotinamide adenine dinucleotide, both pathways converging in the formation of AMP and adenosine via CD73. Among the P1 receptors, only the A2a receptor was significantly upregulated after TAC. T cells isolated from TAC-treated hearts show enhanced production of proinflammatory cytokines (interleukin-3, interleukin-6, interleukin-13, interleukin-17, macrophage inflammatory proteins-1α, and macrophage inflammatory proteins-1β) when CD73 was lacking.
Our data provide first evidence that CD73 on T cells plays an important anti-inflammatory role in TAC-induced heart failure, which is associated with antifibrotic activity and reduced production of proinflammatory cytokines most likely by activation of the adenosine A2a receptor.
心力衰竭发展过程中的结构损伤会导致白细胞浸润增加。由于免疫细胞上的嘌呤能信号可能影响炎症反应，我们使用全身性和T细胞特异性CD73基因敲除小鼠，评估了ecto-5'-核苷酸酶（CD73）在主动脉缩窄（TAC）后心力衰竭发展中的作用。
在16周的时间内，使用荧光激活细胞分选技术，通过多步酶促程序分析浸润衰竭心脏的白细胞。TAC显著增强了白细胞尤其是T细胞的浸润。随着时间的推移，表达CD73的细胞比例仅在细胞毒性T细胞、辅助性T细胞和调节性T细胞上增加。T细胞特异性CD4-CreCD73小鼠的心脏功能显著下降，与全身性CD73基因敲除小鼠中观察到的情况相同，并且与纤维化增强（胶原蛋白、层粘连蛋白、波形蛋白、骨膜蛋白）有关。对从受损心脏分离的T细胞上的细胞外嘌呤降解酶以及P1和P2受体进行定量逆转录聚合酶链反应表达分析，结果显示细胞外三磷酸腺苷和烟酰胺腺嘌呤二核苷酸水解的酶机制显著上调，这两条途径都通过CD73汇聚形成一磷酸腺苷和腺苷。在P1受体中，只有A2a受体在TAC后显著上调。当缺乏CD73时，从TAC处理的心脏中分离的T细胞显示促炎细胞因子（白细胞介素-3、白细胞介素-6、白细胞介素-13、白细胞介素-17、巨噬细胞炎性蛋白-1α和巨噬细胞炎性蛋白-1β）的产生增加。
我们的数据首次证明，T细胞上的CD73在TAC诱导的心力衰竭中起重要的抗炎作用，这可能与抗纤维化活性以及最有可能通过腺苷A2a受体激活而减少促炎细胞因子的产生有关。
