Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan.
Department of Innovative Cancer Immunotherapy, Graduate School of Medicine and Pharmaceutical Sciences (Medicine), University of Toyama, Toyama, Japan.
Front Immunol. 2018 Aug 24;9:1934. doi: 10.3389/fimmu.2018.01934. eCollection 2018.
Regulatory T (Treg) cells are necessary for the maintenance of allogenic pregnancy. However, the repertoire of effector Treg cells at the feto-maternal interface in human pregnancy remains unknown. Our objective was to study T cell receptor (TCR) repertoires of Treg cells during pregnancy compared to normal and complicated pregnancies. Paired samples of peripheral blood and decidua in induced abortion and miscarriage cases were obtained from consenting patients. CD4CD25CD127CD45RA effector Treg cells were single-cell sorted from mononuclear cells. cDNAs of complementarity determining region 3 (CDR3) in TCRβ were amplified from the single cells by RT-PCR and the sequences were analyzed. The TCRβ repertoires were determined by amino acid and nucleotide sequences. Treg cells were classified into clonally expanded and non-expanded populations by CDR3 sequences. We enrolled nine induced abortion cases in the 1st trimester, 12 cases delivered without complications in the 3rd trimester, 11 miscarriages with abnormal chromosomal karyotyped embryo, seven miscarriages with normal chromosomal karyotyped embryo, and seven cases of preeclampsia [median gestational week (interquartile range): 7 (7-9), 39 (38-40), 9 (8-10), 8 (8-10), and 34 (32-37), respectively]. The frequency of clonally expanded populations of effector Treg cells increased in decidua of 3rd trimester cases compared to 1st trimester cases [4.5% (1.4-10.8%) vs. 20.9% (15.4-28.1%), < 0.001]. Clonally expanded Treg cells were rarely seen in peripheral blood. The ratio of clonally expanded populations of decidual effector Treg cells in miscarriages with abnormal and normal embryos was not significantly different compared with that in 1st trimester normal pregnancy. Interestingly, clonally expanded populations of effector Treg cells decreased in preeclampsia compared with that in 3rd trimester normal pregnancy [9.3% (4.4-14.5%) vs. 20.9% (15.4-28.1%), = 0.003]. When repertoires in previous pregnancy and subsequent pregnancy were compared, some portions of the repertoire were shared. TCR repertoires of decidual effector Treg cells are skewed in the 3rd trimester of normal pregnancy. Failure of clonal expansion of populations of decidual effector Treg cells might be related to the development of preeclampsia.
调节性 T（Treg）细胞对于维持同种异体妊娠是必需的。然而，人类妊娠中胎儿-母体界面处效应 Treg 细胞的受体谱仍不清楚。我们的目的是研究妊娠期间与正常和复杂妊娠相比，Treg 细胞的 T 细胞受体（TCR）谱。从同意的患者中获得诱导性流产和自然流产病例的外周血和蜕膜的配对样本。从单核细胞中单细胞分选 CD4+CD25+CD127−CD45RA 效应 Treg 细胞。通过 RT-PCR 从单个细胞中扩增 TCRβ 互补决定区 3（CDR3）的 cDNA，并分析序列。通过氨基酸和核苷酸序列确定 TCRβ 受体谱。通过 CDR3 序列将 Treg 细胞分类为克隆扩增和非扩增群体。我们纳入了 9 例 1 期流产病例，12 例 3 期无并发症分娩病例，11 例胚胎染色体异常流产病例，7 例胚胎染色体正常流产病例和 7 例子痫前期病例[中位数孕龄（四分位距）：7（7-9），39（38-40），9（8-10），8（8-10）和 34（32-37）]。与 1 期妊娠相比，3 期妊娠蜕膜中效应 Treg 细胞的克隆扩增群体的频率增加[4.5％（1.4-10.8％）比 20.9％（15.4-28.1％），<0.001]。外周血中很少见到克隆扩增的 Treg 细胞。与 1 期正常妊娠相比，胚胎染色体异常和正常流产中蜕膜效应 Treg 细胞克隆扩增群体的比率无显著差异。有趣的是，与 3 期正常妊娠相比，子痫前期中效应 Treg 细胞的克隆扩增群体减少[9.3％（4.4-14.5％）比 20.9％（15.4-28.1％），= 0.003]。当比较前次妊娠和后续妊娠的受体谱时，部分受体谱存在共享。正常妊娠 3 期时，蜕膜效应 Treg 细胞的 TCR 受体谱发生偏倚。蜕膜效应 Treg 细胞群体克隆扩增的失败可能与子痫前期的发展有关。
