Trojan Karina, Unterrainer Christian, Weimer Rolf, Bulut Nuray, Morath Christian, Aly Mostafa, Zhu Li, Opelz Gerhard, Daniel Volker
Transplantation-Immunology, Institute of Immunology, University Hospital Heidelberg, Heidelberg, Germany.
Department of Internal Medicine, University of Giessen, Giessen, Germany.
PLoS One. 2017 Mar 15;12(3):e0173773. doi: 10.1371/journal.pone.0173773. eCollection 2017.
There is circumstantial evidence that IFNy+ Treg might have clinical relevance in transplantation. IFNy+ Treg express IFNy receptors and are induced by IFNy. In the present study we investigated in kidney transplant recipients with good long-term stable graft function the absolute cell counts of IFNy+ Treg subsets and whether their expression of Foxp3 is stable or transient.
Helios expression determined by eight-color-fluorescence flow cytometry and methylation status of the Foxp3 Treg specific demethylation region (TSDR) served as indicators for stability of Foxp3 expression. Methylation status was investigated in enriched IFNy+ and IFNy- Treg preparations originating from peripheral blood using high resolution melt analysis. A total of 136 transplant recipients and 52 healthy controls were studied.
Proportions of IFNy+ Treg were similar in patients and healthy controls (0.05% and 0.04% of all CD4+ lymphocytes; p = n.s.). Patients also had similar absolute counts of IFNy producing Helios+ and Helios- Treg (p = n.s.). Most of the IFNy+ and IFNy- Treg in transplant recipients had a methylated Foxp3 TSDR, however, there was a sizeable proportion of IFNy+ and IFNy- Treg with demethylated Foxp3 TSDR. Male and female patients showed more frequently methylated IFNy+ and IFNy- Treg than male and female controls (all p<0.05).
Kidney transplant recipients with good long-term stable graft function have similar levels of IFNy+ Treg as healthy controls. IFNy+ and IFNy- Treg subsets in patients consist of cells with stable and cells with transient Foxp3 expression; however, patients showed more frequently methylated IFNy+ and IFNy- Treg than controls. The data show increased levels of Treg subsets with stable as well as transient Foxp3 expression in patients with stable allograft acceptance compared to healthy controls.
有间接证据表明，IFNγ+调节性T细胞（Treg）可能在移植中具有临床相关性。IFNγ+Treg表达IFNγ受体，并由IFNγ诱导产生。在本研究中，我们调查了具有良好长期稳定移植肾功能的肾移植受者中IFNγ+Treg亚群的绝对细胞计数，以及它们的Foxp3表达是稳定的还是短暂的。
通过八色荧光流式细胞术测定Helios表达以及Foxp3调节性T细胞特异性去甲基化区域（TSDR）的甲基化状态，作为Foxp3表达稳定性的指标。使用高分辨率熔解分析研究源自外周血的富集IFNγ+和IFNγ-Treg制剂中的甲基化状态。共研究了136名移植受者和52名健康对照。
患者和健康对照中IFNγ+Treg的比例相似（分别占所有CD4+淋巴细胞的0.05%和0.04%；p=无统计学差异）。患者产生IFNγ的Helios+和Helios-Treg的绝对计数也相似（p=无统计学差异）。移植受者中大多数IFNγ+和IFNγ-Treg的Foxp3 TSDR呈甲基化状态，然而，有相当比例的IFNγ+和IFNγ-Treg的Foxp3 TSDR呈去甲基化状态。男性和女性患者中IFNγ+和IFNγ-Treg甲基化的频率高于男性和女性对照（所有p<0.05）。
具有良好长期稳定移植肾功能的肾移植受者中IFNγ+Treg水平与健康对照相似。患者中的IFNγ+和IFNγ-Treg亚群由Foxp3表达稳定和短暂的细胞组成；然而，患者中IFNγ+和IFNγ-Treg甲基化的频率高于对照。数据显示，与健康对照相比，移植稳定受者中Foxp3表达稳定和短暂的调节性T细胞亚群水平均升高。
