Danish Multiple Sclerosis Center, Copenhagen University Hospital, Rigshospitalet, Glostrup, Denmark.
Department of Neurology, Turku University Hospital, Turku, Finland.
Front Immunol. 2021 Jan 28;11:552101. doi: 10.3389/fimmu.2020.552101. eCollection 2020.
Pregnancy affects the disease course in multiple sclerosis (MS), particularly in the third trimester, where the relapse rate is reduced by as much as two thirds. This study aimed at identifying changes in microRNA (miRNA) and immune cell phenotypes in pregnant MS patients. Discovery and validation studies to detect differentially expressed miRNAs were performed with quantitative real-time PCR on peripheral blood mononuclear cells (PBMC). Flow cytometry analysis was performed on PBMC stained with antibodies directed against surface markers of antigen presenting cells (APCs), NK-cells, NKT cells, CD4+ and CD8+ T cells and subsets of these cell types, including PDL1 and PDL2 expressing subsets. RNA was extracted from whole blood, monocytes, and NK-cells to investigate expression and correlation between regulated miRNAs and mRNAs. In total, 15 miRNAs were validated to be differentially expressed between third trimester pregnant and postpartum MS patients (Benjamini-Hochberg false discovery rate from p = 0.03-0.00004). Of these, 12 miRNAs were downregulated in pregnancy and 6 of the 15 miRNAs were altered by more than ±2-fold (+2.99- to -6.38-fold). Pregnant MS patients had a highly significant increase in the percentage of monocytes and a decrease of NK-cells and myeloid dendritic cells compared to non-pregnant MS patients. We confirm previous reports of a relative increase in CD56-bright NK-cells and a decrease in CD56-dim NK-cells in third trimester of pregnancy and report an increase in non-committed follicular helper cells. and expression was increased in pregnant patients together with . Also, in monocytes , , and were upregulated whereas miR-1, miR-20a, miR-28, miR-95, miR-146a, miR-335, and miR-625 were downregulated between pregnant and untreated MS patients. , , and were predicted targets of MS pregnancy-changed miRNAs, further supported by their negative correlations. Additionally, previously identified pregnancy-regulated mRNAs were identified as predicted targets of the miRNAs. PDL1 and PDL2 bind PD-1 expressed on T cells with an inhibitory effect on T-cell proliferation and increase in IL10 production. These results indicate that some of the effects behind the disease-ameliorating third trimester of pregnancy might be caused by changed expression of miRNAs and immunoregulatory molecules in monocytes.
妊娠会影响多发性硬化症（MS）的疾病进程，尤其是在妊娠晚期，复发率降低了多达三分之二。本研究旨在确定妊娠 MS 患者中微小 RNA（miRNA）和免疫细胞表型的变化。通过定量实时 PCR 在外周血单核细胞（PBMC）上进行发现和验证研究，以检测差异表达的 miRNA。使用针对抗原呈递细胞（APC）、NK 细胞、NKT 细胞、CD4+和 CD8+T 细胞以及这些细胞类型的亚群表面标记的抗体对 PBMC 进行流式细胞术分析，包括表达 PDL1 和 PDL2 的亚群。从全血、单核细胞和 NK 细胞中提取 RNA，以研究调节 miRNA 和 mRNAs 之间的表达和相关性。总共验证了 15 种 miRNA 在第三孕期妊娠和产后 MS 患者之间的差异表达（Benjamini-Hochberg 假发现率从 p = 0.03-0.00004）。其中，12 种 miRNA 在妊娠时下调，15 种 miRNA 中有 6 种变化超过±2 倍（+2.99-至-6.38 倍）。与非妊娠 MS 患者相比，妊娠 MS 患者的单核细胞百分比显著升高，NK 细胞和髓样树突状细胞减少。我们证实了之前关于第三孕期妊娠中 CD56-bright NK 细胞相对增加和 CD56-dim NK 细胞减少的报告，并报告了非定型滤泡辅助细胞的增加。在妊娠患者中，表达增加，而在非妊娠 MS 患者中，表达减少。此外，在单核细胞中，miR-1、miR-20a、miR-28、miR-95、miR-146a、miR-335 和 miR-625 的表达下调，而 miR-1、miR-20a、miR-28、miR-95、miR-146a、miR-335 和 miR-625 的表达下调。、和是 MS 妊娠相关 miRNA 的预测靶点，进一步得到其负相关关系的支持。此外，还鉴定了先前鉴定的妊娠调节 mRNA 是 miRNA 的预测靶点。PDL1 和 PDL2 与 T 细胞上表达的 PD-1 结合，抑制 T 细胞增殖并增加 IL10 产生。这些结果表明，妊娠晚期疾病缓解的一些影响可能是由单核细胞中 miRNA 和免疫调节分子表达的变化引起的。
