摘要
目的本研究拟对体外诱导的Helios+调节性T细胞(induced Helios+ regulatory T cell,Helios+ iTreg)的特征进行分析,并比较其与正常人外周血固有Helios+ Treg(natural Helios+ Treg, Helios+ nTreg)的差异。方法采集并分选正常人外周CD4+CD25-效应性T细胞(effector T cell,Teff),体外以抗CD3/抗CD28合并IL-2及TGF-β刺激诱导Helios+ iTreg生成,以外周血Helios+ nTreg作为对照,通过流式细胞术和real-time PCR分别分析其表型特征Foxp3+ Treg特异性去甲基化区(Treg-specific demethylated region,TSDR)的甲基化水平。结果体外诱导的Helios+ iTreg高表达CD45RA,而Helios+ nTreg几乎不表达CD45RA;体外诱导的Helios+ iTreg较Helios+ nTreg CD127、ICOS和PD-1的水平明显升高,IL-2和IFN-γ的表达水平明显升高,但CXCR3、CCR4等趋化因子受体的水平则明显下降;另外CD25的表达水平也明显下降;诱导型Helios+ iTreg TSDR的甲基化水平明显低于Helios+ nTreg。结论诱导型Helios+ iTreg具有与Helios+ nTreg不同的特征,提示其在生理及病理条件下可能发挥不同的免疫调节作用。
Objective To investigate the phenotypic and functional properties of in vitro-induced Helios+ regulatory T cells (Helios+ iTreg) and to analyze the differences between Helios+ iTreg and Helios+ thymic-derived natural Treg (Helios+ nTreg).MethodsCD4+ CD25- effector T cells (CD4+ CD25- Teff) that were separated from healthy subjects were cultured with anti-CD3 and anti-CD28 antibodies and stimulated with IL-2 and TGF-β to induce the generation of Helios+ iTreg. Flow cytometry and real-time PCR were respectively used to analyze the differences in phenotype and CpG methylation in Treg-specific demethylated region (TSDR) between Helios+ iTreg and Helios+ nTreg derived from the same donor.ResultsCD45RA was highly expressed on Helios+ iTreg, but not on Helios+ nTreg. Additionally, Helios+ iTreg expressed significantly higher levels of CD127, ICOS and PD-1, but lower levels of CXCR3, CCR4 and CD25 than Helios+ nTreg did. IFN-γ and IL-2 that expressed by Helios+ iTreg were more than those by Helios+ nTreg. It was also found that the level of CpG methylation in TSDR was much higher in Helios+ iTreg than in Helios+ nTreg.ConclusionBoth nTreg and iTreg expressed Helios, but the phenotypic and functional properties of the two Treg subsets were different. It was suggested that Helios+ iTreg and Helios+ nTreg might play different roles in the immune system.
作者
杨帆
范臻佳
顾燕英
吴蓓颖
林琳
蔡刚
Yang Fan;Fan Zhenjia;Gu Yanying;Wu Belying;Lin Lin;Cai Gang(Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shang- hai 200025, Chin)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2018年第4期241-247,共7页
Chinese Journal of Microbiology and Immunology