Newly characterized decidual Tim-3+. Treg cells are abundant during early pregnancy and driven by IL-27 coordinately with Gal-9 from trophoblasts
Newly characterized decidual Tim-3+. Treg cells are abundant during early pregnancy and driven by IL-27 coordinately with Gal-9 from trophoblasts
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DOI：
10.1093/humrep/deaa223
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发表时间：
2020-11-01
影响因子：
6.1
通讯作者：
Liao, Aihua
中科院分区：
文献类型：
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作者：
Hu, Xiaohui;Zhu, Qian;Liao, Aihua
研究问题：早孕蜕膜中TIM-3(+)调节性T(Treg)细胞积聚的机制是什么？它的破坏是否与反复妊娠丢失(RPL)有关？摘要：滋养细胞分泌的IL-27和Gal-9激活了CD4(+)T细胞和Treg细胞中的Tim-3信号通路，从而促进了TIM-3+Treg细胞的积聚，IL-27和Gal-9的异常表达与RPL患者免疫耐受受损有关。已知：Tim-3(+)Treg细胞更好地抑制Tef细胞的增殖，并且表现出比TIM-3Treg细胞更高的增殖活性。TIM-3(+)Treg细胞是许多肿瘤中T细胞功能障碍的组织特异性促进剂。这些细胞表达一种影响和塑造肿瘤微环境的独特因素。研究设计、大小、持续时间：动物研究包括80只正常怀孕的小鼠。在人体研究中，采集了32名正常孕妇和23名RPL患者的早孕蜕膜组织，用于流式细胞仪分析。采集15例正常孕妇胎盘组织进行免疫组织化学分析。对5例正常孕妇、5例RPL患者和5例流产妇女的胎盘组织进行免疫印迹分析。体外实验采自30名正常孕妇的血样。本研究在2017年1月至2019年3月期间进行。材料、背景、方法：在本研究中，我们利用流式细胞术研究了TIM-3(+)CD4(+)T细胞在整个妊娠过程中的积聚动力学，以及TIM-3(+)Treg细胞在整个妊娠过程中的比例。我们比较了正常妊娠期间蜕膜CD4(+)T细胞和Treg细胞上Tim-3的表达与RPL患者同一细胞群上的表达。RT-PCR法检测胎盘组织中IL-27和Gal-9的转录水平，免疫印迹法检测胎盘组织中Gal-9蛋白的表达。体外共培养模型由妊娠早期的外周血CD4(+)T细胞和原代滋养层细胞组成，用于模拟母胎环境。MAIN结果和机会的作用：在小鼠子宫中存在的Tim-3(+)Treg细胞的百分比随着妊娠的进行而波动，但在脾中不变。子宫中TIM3(+)Treg细胞的水平在妊娠第6.5天(E6.5)达到峰值，然后逐渐下降，到E18.5下降到未怀孕的水平。在妊娠小鼠中，TIM-3(+)Treg细胞占子宫Treg细胞的40%-70%，但在脾中的丰度要低得多。约60%的蜕膜Treg细胞在E6.5表达TIM-3。在这些蜕膜TIM3(+)Treg细胞中，近90%的细胞PD-I阳性。然而，只有约16%的TIM3+Treg细胞表达PD-I。阻断TIM-3信号通路可降低Treg细胞比例，导致胚胎吸收。此外，在妊娠6-9周患有RPL的妇女中，与正常妊娠妇女相比，TIM-3的表达水平显著降低。在正常妊娠中，蜕膜CD4(+)T细胞上TIM-3的表达最初是由IL-27诱导的。Gal-9-Tim-3相互作用促进蜕膜Tim-3(+)、CD4(+)T细胞向Treg细胞分化。IL-27和Gal-9在体外协同诱导TIM-3(+)Treg细胞。规模数据：N/ALIMITATIONS，警告理由：由于中期和晚期蜕膜的可获得性有限，我们没有研究人类蜕膜TIM-3(+)CD4(+)T和TIM-3(+)Treg细胞群在整个妊娠过程中的动力学。此外，由于蜕膜中TIM-3(+)Treg细胞的数量有限且数量较少，因此缺乏关于这些细胞的功能抑制数据。研究结果的广泛意义：这些发现可能对RPL具有临床治疗意义。
STUDY QUESTION: What is the mechanism of Tim-3(+) regulatory T (Treg)-cell accumulation in the decidua during early pregnancy and is its disruption associated with recurrent pregnancy loss (RPL)?SUMMARY ANSWER: IL-27 and Gal-9 secreted by trophoblasts activate the Tim-3 signaling pathway in CD4(+) T cells and Treg cells and so promote accumulation of Tim-3+ Treg cells, the abnormal expression of IL-27 and Gal-9 is associated with impaired immunologic tolerance in RPL patients.WHAT IS KNOWN ALREADY: Tim-3(+) Treg cells are better suppressors of Teff cell proliferation, and display higher proliferative activity than Tim-3 Treg cells. Tim-3(+) Treg cells are tissue-specific promoters of T-cell dysfunction in many tumors. These cells express a unique factor that influences and shapes the tumor microenvironment.STUDY DESIGN, SIZE, DURATION: The animal study included 80 normal pregnant mice. In human study, decidua tissues in the first trimester for flow cytometry analysis were collected from 32 normal pregnant women and 23 RPL patients. Placenta tissues for immunohistochemistry analysis were collected from 15 normal pregnant women. Placenta tissues for western blot analysis were collected from 5 normal pregnant women, 5 RPL patients and 5 women who have experienced one miscarriage. Blood samples for in vitro experiments were collected from 30 normal pregnant women. This study was performed between January 2017 and March 2019.PARTICIPANTS/MATERIALS, SETTING, METHODS: In this study, we investigated the kinetics of Tim-3(+) CD4(+) T-cell accumulation, and the proportions of Tim-3(+) Treg cells throughout murine pregnancies using flow cytometry. We compared Tim-3 expression on decidual CD4(+) T cells and Treg cells during normal pregnancies with expression on the same cell populations in women suffering from RPL. IL-27 and Gal-9 transcription and protein expression in the placenta were determined by RT-PCR and western blot, respectively. An in vitro co-culture model consisting of peripheral CD4(+) T cells and primary trophoblasts from early pregnancy was used to mimic the maternal-fetal environment.MAIN RESULTS AND THE ROLE OF CHANCE: The percentage of Tim-3(+) Treg cells present in mouse uteri fluctuates as gestation proceeds but does not change in the spleen. Levels of Tim3(+) Treg cells in uteri peaked at pregnancy Day 6.5 (E 6.5), then progressively diminished, and fell to non-pregnant levels by E18.5. In pregnant mice, Tim-3(+) Treg cells constituted 40-70% of Treg cells in uteri but were present at much lower abundance in spleens. About 60% of decidual Treg cells were Tim-3 positive at E6.5. Of these decidual Tim3(+) Treg cells, nearly 90% were PD- I positive. However, only about 16% of Tim3 + Treg cells expressed PD-I. Blocking the Tim-3 signaling pathway decreased the proportion of Treg cells and led to embryo resorption. Moreover, much lower Tim-3 expression was observed CD4(+) T cells and Treg cells in women who had suffered from RPL at 6-9 gestational weeks compared with those who had normal pregnancies at matched gestations. In a normal pregnancy, Tim-3 expression on decidual CD4(+) T cells is induced initially by IL-27. Then Gal-9-Tim-3 interaction promotes differentiation of decidual Tim-3(+) CD4(+) T cells into Treg cells. IL-27 and Gal-9 cooperatively induced Tim-3(+) Treg cells in vitro.LARGE SCALE DATA: N/ALIMITATIONS, REASONS FOR CAUTION: We did not investigate the kinetics of human decidual Tim-3(+) CD4(+) T and Tim-3(+) Treg cell populations throughout pregnancy due to limited availability of second and third trimester decidua. In addition, functional suppressive data on the decidual Tim-3(+) Treg cells are lacking due to limited and low quantities of these cells in decidua.WIDER IMPLICATIONS OF THE FINDINGS: These findings might have therapeutic clinical implications in RPL.