Abstract
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 vivo 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.
Highlights
Spontaneous miscarriage is the most common complication of early pregnancy
Since Medawar and colleagues introduced the concept of immunological tolerance between a mother and her fetus [27], and Billingham theorised the breakdown of this tolerance underlying the pregnancy loss [28], a number of studies have investigated the roles of specific immune cells and molecules in the etiology of Recurrent spontaneous miscarriage (RM)
There have been plenty of studies focussing on this type 1/type 2 shift phenomenon in T helper (Th) cells and dNK cells, the present study showed an overall decidual Th1/Th2 and decidual NK1 (dNK1)/decidual NK2 (dNK2) profile that validated the Th and dNK type1/type2 shift simultaneously for the first time
Summary
Spontaneous miscarriage is the most common complication of early pregnancy. Recurrent spontaneous miscarriage (RM), defined by two or more failed pregnancies, occurs in 1–2% of fertile women [1,2]. Present knowledge about the etiology of RM mainly includes parental or fetal chromosomal defects, anatomical anomalies, endocrine disorders, infections, and maternal autoimmune factors [3,4]. A considerable number of trials targetting the feto–maternal interface have been carried out for decades, including theoretical models covering alloimmunity, autoimmunity, and the innate immune system. Both a T helper (Th) cell imbalance and a natural killer (NK) cell-mediated immunity contribute to the unknown mechanisms underlying unexplained recurrent miscarriage (URM) cases [5-8]
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