Jiang Lai, Tang Chaoliang, Gong Yanping, Liu Yujie, Rao Jie, Chen Suyu, Qu Wanjun, Wu Dabao, Lei Lei, Chen Ling
The First Affiliated Hospital, Department of Obstetrics and Gynecology, University of Science and Technology of China, Hefei, China.
The First Affiliated Hospital, Department of Anesthesiology, University of Science and Technology of China, Hefei, China.
Braz J Med Biol Res. 2018;51(8):e7334. doi: 10.1590/1414-431x20187334. Epub 2018 May 28.
Pregnancy-induced hypertension (PIH) causes significant maternal and fetal morbidity and mortality. A decreased number of regulatory T (Treg) cells is associated with the pathogenesis of PIH. The programmed cell death-1 (PD-1)/PD-ligand 1 (PD-L1) pathway is critical to normal pregnancy (NP) by promoting Treg cell development. However, the relationship between PD-1/PD-L1 and Treg differentiation in PIH has not been fully elucidated. In this study, venous blood was obtained from 20 NP and 58 PIH patients. Peripheral blood mononuclear cells (PBMCs) were isolated from venous blood. The levels of Treg-related cytokines (TGF-β, IL-10, and IL-35) in serum and PBMCs were measured by ELISA. The percentage of Treg cells in PBMCs was assessed by flow cytometry. The mRNA levels of Treg-specific transcription factor Foxp3 in PBMCs, and PD-1 and PD-L1 in Treg cells were detected by qRT-PCR. The protein levels of PD-1 and PD-L1 in Treg cells were evaluated by western blot. The serum levels of TGF-β, IL-10, IL-35, and Foxp3 mRNA expression and CD4+CD25+ Treg cell percentage in PBMCs were decreased in PIH. Furthermore, a significant increase of PD-1 in Treg cells was found in PIH compared with NP. In addition, PD-L1 Fc, an activator of PD-1/PD-L1 pathway, increased Treg cell percentage, enhanced Foxp3 mRNA expression, and elevated levels of TGF-β, IL-10, and IL-35 in PBMCs. However, anti-PD-L1 mAb exerted a reverse effect. These findings revealed that PD-L1 Fc had a favorable effect on Treg cell differentiation, indicating a potential therapeutic value of PD-1/PD-L1 pathway for PIH treatment.
妊娠期高血压疾病（PIH）会导致严重的母婴发病和死亡。调节性T（Treg）细胞数量减少与PIH的发病机制有关。程序性细胞死亡蛋白1（PD-1）/程序性死亡配体1（PD-L1）通路通过促进Treg细胞发育对正常妊娠（NP）至关重要。然而，PIH中PD-1/PD-L1与Treg分化之间的关系尚未完全阐明。在本研究中，从20例NP患者和58例PIH患者中采集静脉血。从静脉血中分离外周血单个核细胞（PBMC）。采用酶联免疫吸附测定法（ELISA）检测血清和PBMC中Treg相关细胞因子（转化生长因子-β、白细胞介素-10和白细胞介素-35）的水平。通过流式细胞术评估PBMC中Treg细胞的百分比。采用实时定量聚合酶链反应（qRT-PCR）检测PBMC中Treg特异性转录因子叉头框蛋白3（Foxp3）的mRNA水平以及Treg细胞中PD-1和PD-L1的mRNA水平。通过蛋白质免疫印迹法评估Treg细胞中PD-1和PD-L1的蛋白水平。PIH患者血清中转化生长因子-β、白细胞介素-10、白细胞介素-35水平以及PBMC中Foxp3 mRNA表达和CD4+CD25+ Treg细胞百分比均降低。此外，与NP相比，PIH患者Treg细胞中PD-1显著增加。此外，PD-1/PD-L1通路激活剂PD-L1 Fc增加了Treg细胞百分比，增强了Foxp3 mRNA表达，并提高了PBMC中转化生长因子-β、白细胞介素-10和白细胞介素-35的水平。然而，抗PD-L1单克隆抗体则产生相反的效果。这些发现表明，PD-L1 Fc对Treg细胞分化具有有益作用，提示PD-1/PD-L1通路在PIH治疗中具有潜在的治疗价值。
