■为了研究蜕膜自然杀伤(NK)细胞和γδT细胞的特征性功能变化,蜕膜中的两个免疫细胞,在体外受精-胚胎移植(IVF-ET)妊娠的母胎界面。
■从12名自然妊娠(NP)妇女和32名IVF-ET妊娠妇女中收集了蜕膜样本,纳入NP组和IVF-ET组,分别。然后将部分蜕膜样本石蜡包埋,进行HE染色和免疫荧光染色,而其余样品被消化并且Percoll用于通过梯度离心分离蜕膜免疫细胞(DIC)。采用流式细胞术检测蜕膜NK细胞和γδT细胞的细胞计数及其表面活化标志物的表达水平。NP和IVF-ET组中的CD69和NKG2D。此外,IFN-γ的表达水平,TNF-α,IL-17A,和IL-10,细胞内细胞因子,和颗粒酶B,穿孔素,和颗粒素,细胞溶解颗粒,被测量。对比分析相关免疫学指标的特征性变化。
■组织标本的HE染色显示,观察到蜕膜的典型结构,淋巴细胞在蜕膜中富集。免疫荧光染色显示,IVF-ET组有核细胞中蜕膜NK(dNK)细胞的百分比明显低于NP组(P<0.05)。流式细胞术分析DIC显示,与NP组相比,IVF-ET组dNK细胞百分比降低(P<0.05),IL-10和穿孔素的表达水平明显降低(P<0.05)。然而,两组蜕膜γδT(dγδT)细胞计数无明显差异。IL-10、IL-17A、IVF-ET组穿孔素表达下调(P<0.05)。IFN-γ的表达无显著差异,TNF-α,颗粒酶B,和颗粒素,细胞功能指标(P>0.05)。
■IVF-ET妊娠妇女的dNK细胞计数和dNK和dγδT细胞的一些细胞内细胞因子的分泌有一定程度的下降,这表明母胎界面的免疫微环境可能发生了某些变化。这些变化对妊娠结局的具体影响需要进一步调查。
UNASSIGNED: To investigate the characteristic functional changes of the decidual natural killer (NK) cells and γδ T cells, two immunocytes in the decidua, at the maternal-fetal interface in in vitro fertilization-embryo transfer (IVF-ET) pregnancy.
UNASSIGNED: Decidual samples were collected from 12 women of natural pregnancy (NP) and 32 women of IVF-ET pregnancy, who were enrolled in the NP group and the IVF-ET group, respectively. Then part of the decidual samples were paraffin-embedded for HE staining and immunofluorescence staining, while the rest of the samples were digested and Percoll was used for isolating decidual immunocytes (DICs) by gradient centrifugation. Flow cytometry was used to determine the cell counts of decidual NK cells and γδ T cells and the expression levels of their surface activation markers, CD69 and NKG2D in the NP and the IVF-ET groups. In addition, the expression levels of IFN-γ, TNF-α, IL-17A, and IL-10, the intracellular cytokines, and granzyme B, perforin, and granulysin, the cytolytic granules, were measured. The characteristic changes in the relevant immunological indicators were compared and analyzed.
UNASSIGNED: HE staining of the tissue specimens showed that the typical structure of decidua was observed, and that lymphocytes were enriched in the decidua. Immunofluorescence staining showed that the percentage of decidual NK (dNK) cells in nucleated cells of the IVF-ET group was significantly lower than that of the NP group ( P<0.05). Flow cytometry analysis of DICs showed that, compared with those of the NP group, the percentage of dNK cells of the IVF-ET group was decreased ( P<0.05) and the expression levels of IL-10 and perforin were significantly decreased in the IVF-ET group ( P<0.05). However, there was no significant difference in the decidual γδ T (dγδT) cell count between the two groups. The expression of IL-10, IL-17A, and perforin was downregulated in the IVF-ET group ( P<0.05). There was no significant difference in the expression of IFN-γ, TNF-α, granzyme B, and granulysin, the cellular function indicators ( P>0.05).
UNASSIGNED: The dNK cell count and the secretion of some intracellular cytokines of dNK and dγδT cells of women of IVF-ET pregnancy decreased to some degree, which suggests that certain changes may have taken place in the immunological microenvironment at the maternal-fetal interface. The specific effect of these changes on pregnancy outcomes needs further investigation.