Dong Peng, Wen Xi, Liu Jia, Yan Cui-Yan, Yuan Jing, Luo Lan-Rong, Hu Qiao-Fei, Li Jian
Department of Obstetrics and Gynecology, Xuanwu Hospital, Capital Medical University, Beijing, China.
Family Planning Department, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
Biosci Rep. 2017 May 19;37(3). doi: 10.1042/BSR20170150. Print 2017 Jun 30.
Th1/Th2 imbalance is considered as a mechanism for recurrent miscarriage. The NK1/NK2 paradigm is hypothesised to play an important role in pregnancy. However, few results showed simultaneous changes of these subsets in decidual tissues. The present study aimed to detect the decidual mononuclear cells (dMo), and the Th1/Th2, and NK1/NK2 paradigm simultaneously using multiparametric flow cytometry (MFC) in unexplained recurrent miscarriages (URM). Mononuclear cells were isolated from the decidual tissues of URM cases and early pregnant women. The mononuclear cell percent was demonstrated by detecting the expression of CD3, CD4, CD8, CD56, and CD16 extracellular markers, interferon (IFN)-γ, and interleukin (IL)-4 intracellular markers in live cells using 8-color flow cytometry with forward scatter (FSC)/side scatter (SSC) and FSC/viability (Vt) initial gating strategies, and the ratios of Th1/Th2 and decidual NK1 (dNK1)/decidual NK2 (dNK2) cells were compared between the subject groups. Two initial gating strategies of the FSC/SSC or FSC/Vt, with central or extended gating scales, were adapted, and there was no main effect or interaction for the cell proportions, except for the type 1 and type 2 subsets in the FSC/Vt extended gating strategy. There was no significant difference of the proportions of the decidual T, dNK, NKT-like, Th, and Tc cells between the two groups. However, the Th1/Th2 and dNK1/dNK2 ratios in the URM patients were higher compared with the normal group when using the FSC/Vt extended gating strategy. The present study provides means to detect Th1/Th2 and dNK1/dNK2 simultaneously in URM patients for large sample investigations in the future.
Th1/Th2失衡被认为是复发性流产的一种机制。NK1/NK2模式被假设在妊娠中起重要作用。然而，很少有结果显示这些亚群在蜕膜组织中同时发生变化。本研究旨在利用多参数流式细胞术（MFC）同时检测不明原因复发性流产（URM）患者的蜕膜单核细胞（dMo）、Th1/Th2以及NK1/NK2模式。从URM病例和早孕妇女的蜕膜组织中分离单核细胞。使用具有前向散射（FSC）/侧向散射（SSC）和FSC/活力（Vt）初始门控策略的8色流式细胞术，通过检测活细胞中细胞外标志物CD3、CD4、CD8、CD56和CD16、细胞内标志物干扰素（IFN）-γ和白细胞介素（IL）-4的表达来确定单核细胞百分比，并比较各研究组之间Th1/Th2和蜕膜NK1（dNK1）/蜕膜NK2（dNK2）细胞的比例。采用了FSC/SSC或FSC/Vt的两种初始门控策略，门控尺度为中心或扩展型，除了FSC/Vt扩展门控策略中的1型和2型亚群外，细胞比例没有主要效应或相互作用。两组之间蜕膜T细胞、dNK细胞、NKT样细胞、Th细胞和Tc细胞的比例没有显著差异。然而，当使用FSC/Vt扩展门控策略时，URM患者的Th1/Th2和dNK1/dNK2比值高于正常组。本研究为未来在URM患者中同时检测Th1/Th2和dNK1/dNK2提供了方法，以便进行大样本研究。
