Immune cell and transcriptomic analysis of the human decidua in term and preterm parturition.
Immune cell and transcriptomic analysis of the human decidua in term and preterm parturition.
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DOI：
10.1093/molehr/gax038
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发表时间：
2017-10-01
影响因子：
4
通讯作者：
Norman JE
中科院分区：
文献类型：
--
作者：
Rinaldi SF;Makieva S;Saunders PT;Rossi AG;Norman JE
足月和早产的分娩是否与蜕膜淋巴细胞密度的改变和蜕膜转录组的广泛变化有关？足月和早产的分娩开始与蜕膜中广泛的基因表达变化有关，其中许多与炎症信号传导有关，但与所检查的任何蜕膜淋巴细胞群数量的变化无关。鉴于其直接位于母胎界面的位置，蜕膜很可能在分娩开始时发挥关键作用，然而，在足月和早产时，蜕膜中发生的与临产开始相关的分子事件仍然相对不明确。本研究旨在利用流式细胞术和微阵列分析来研究与分娩开始相关的蜕膜免疫细胞环境的变化，并确定与足月和早产 (PTL) 相关的蜕膜基因特征。这项研究使用了从 4 个临床组的 36 名女性身上收集的蜕膜样本：足月（妊娠 38-42 周）、未临产、TNL；劳动期限，TL；早产（妊娠<35周）未临产，PTNL；和早产，PTL。从四个患者组中收集的新鲜蜕膜组织中分离蜕膜淋巴细胞，并用一组抗体（CD45、CD3、CD19、CD56、CD4、CD8 和 TCRVα24-Jα18）染色，以研究蜕膜中存在的淋巴细胞群（TNL，n = 8；TL，n = 7；PTNL，n = 5；PTL，n = 5）。从蜕膜组织中提取 RNA，并进行 Illumina HT-12v4.0 BeadChip 表达微阵列分析（TNL，n = 11；TL，n = 8；PTNL，n = 7；PTL，n = 10）。使用定量实时 PCR (qRT-PCR) 来验证微阵列结果。蜕膜淋巴细胞（T 细胞、NK 细胞、B 细胞和不变自然杀伤 (iNKT) 细胞）的相对比例不受妊娠或分娩状态的影响。然而，我们发现 PTL 蜕膜样本中非经典 MHC 蛋白 CD1D 的表达升高（P < 0.05），这表明 PTL 中蜕膜不变 NKT (iNKT) 细胞的活化可能增加。 term 和 PTL 都与广泛的基因表达变化相关，特别是与炎症信号传导相关。 qRT-PCR 分析证实了 TL（IL-6、PTGS2、ATF3、IER3 和 TNFAIP3）和 PTL（CXCL8、MARCO、LILRA3 和 PLAU）中候选基因的上调。微阵列数据可在登录号 E-MTAB-5353 中获得。虽然我们的患者样本中没有观察到淋巴细胞数量的变化，但我们没有调查所检查的任何免疫细胞亚群的激活状态，因此，这些细胞的功能可能会因分娩开始而改变。此外，我们的转录组分析结果是描述性的，在现阶段，我们无法证明与所检查的任何基因的上调以及足月或 PTL 的开始之间存在直接因果关系。我们的研究结果表明，分娩的开始与蜕膜转录组的广泛变化相关，并且与足月和 PTL 相关的明显基因表达变化。我们证实蜕膜内存在炎症特征，并且我们还报告了参与调节炎症反应的几个基因的上调。鉴定参与调节炎症反应的基因可能为开发新的、更有效的预防早产（PTB）疗法提供新的分子靶标。这些目标是迫切需要的。这项工作得到了医学研究委员会（拨款号 MR/L002657/1）和婴儿慈善机构 Tommy's 的支持。在此项目期间，Jane Norman 获得了 Tommy's 慈善机构和国家卫生研究院 PTB 的研究资助。简·诺曼 (Jane Norman) 也是葛兰素史克 (GSK) 数据监测委员会的成员，负责一项预防肺结核的研究，她的机构因此获得了经济补偿。其他作者没有任何需要声明的利益冲突。
Is labour, both at term and preterm, associated with alterations in decidual lymphocyte densities and widespread changes to the decidual transcriptome? The onset of parturition, both at term and preterm, is associated with widespread gene expression changes in the decidua, many of which are related to inflammatory signalling, but is not associated with changes in the number of any of the decidual lymphocyte populations examined. Given its location, directly at the maternal–foetal interface, the decidua is likely to play a pivotal role in the onset of parturition, however, the molecular events occurring in the decidua in association with the onset of labour, both at term and preterm, remain relatively poorly defined. Using flow cytometry and microarray analysis, the present study aimed to investigate changes to the immune cell milieu of the decidua in association with the onset of parturition and define the decidual gene signature associated with term and preterm labour (PTL). This study used decidual samples collected from 36 women across four clinical groups: term (38–42 weeks of gestation) not in labour, TNL; term in labour, TL; preterm (<35 weeks of gestation)not in labour, PTNL; and preterm in labour, PTL. Decidual lymphocytes were isolated from fresh decidual tissue collected from women in each of our four patient groups and stained with a panel of antibodies (CD45, CD3, CD19, CD56, CD4, CD8 and TCRVα24-Jα18) to investigate lymphocyte populations present in the decidua (TNL, n = 8; TL, n = 7; PTNL, n = 5; PTL, n = 5). RNA was extracted from decidual tissue and subjected to Illumina HT-12v4.0 BeadChip expression microarrays (TNL, n = 11; TL, n = 8; PTNL, n = 7; PTL, n = 10). Quantitative real-time PCR (qRT-PCR) was used to validate the microarray results. The relative proportions of decidual lymphocytes (T cells, NK cells, B cells and invariant natural killer (iNKT) cells) were unaffected by either gestation or labour status. However, we found elevated expression of the non-classical MHC-protein, CD1D, in PTL decidua samples (P < 0.05), suggesting the potential for increased activation of decidual invariant NKT (iNKT) cells in PTL. Both term and PTL were associated with widespread gene expression changes, particularly related to inflammatory signalling. Up-regulation of candidate genes in TL (IL-6, PTGS2, ATF3, IER3 and TNFAIP3) and PTL (CXCL8, MARCO, LILRA3 and PLAU) were confirmed by qRT-PCR analysis. Microarray data are available at under accession number E-MTAB-5353. Whilst no changes in lymphocyte number were observed across our patient samples, we did not investigate the activation state of any of the immune cell sub-populations examined, therefore, it is possible that the function of these cells may be altered in association with labour onset. Additionally, the results of our transcriptomic analyses are descriptive and at this stage, we cannot prove direct causal link with the up-regulation of any of the genes examined and the onset of either term or PTL. Our findings demonstrate that the onset of parturition is associated with widespread changes to the decidual transcriptome, and there are distinct gene expression changes associated with term and PTL. We confirmed that an inflammatory signature is present within the decidua, and we also report the up-regulation of several genes involved in regulating the inflammatory response. The identification of genes involved in regulating the inflammatory response may provide novel molecular targets for the development of new, more effective therapies for the prevention of preterm birth (PTB). Such targets are urgently required. This work was supported by Medical Research Council (grant number MR/L002657/1) and Tommy's, the baby charity. Jane Norman has had research grants from the charity Tommy's and from the National Institute for Health Research on PTB during the lifetime of this project. Jane Norman also sits on a data monitoring committee for GSK for a study on PTB prevention and her institution receives financial recompense for this. The other authors do not have any conflicts of interest to declare.
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DOI：
10.1136/bmj.e6226
发表时间：
2012-10-09
期刊：
BMJ (Clinical research ed.)
影响因子：
--
作者：
Haas DM;Caldwell DM;Kirkpatrick P;McIntosh JJ;Welton NJ
通讯作者：
Welton NJ
影响因子：
3.1
作者：
Brennan DJ;McGee SF;Rexhepaj E;O'Connor DP;Robson M;O'Herlihy C
通讯作者：
O'Herlihy C
影响因子：
3.4
作者：
Gomez-Lopez, N.;Estrada-Gutierrez, G.;Vadillo-Ortega, R.
通讯作者：
Vadillo-Ortega, R.
影响因子：
19.6
作者：
BARICOS, WH;CORTEZ, SL;SCHNAPER, HW
通讯作者：
SCHNAPER, HW
影响因子：
3.8
作者：
Keelan, JA;Blumenstein, M;Mitchell, MD
通讯作者：
Mitchell, MD