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[体外受精-胚胎移植妊娠母胎界面免疫微环境特征变化的实验研究]

[Experimental Study on the Characteristic Changes of the Immunological Microenvironment at the Maternal-Fetal Interface in IVF-ET Pregnancy].

作者信息

Liu Yuan, Li Li-Man, Chen Hong-Qin, Feng Ting, Zhou Wen-Jie, Liu Ying, Zhou Rong, Li Hong

机构信息

Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu 610041, China.

Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2023 Mar;54(2):350-356. doi: 10.12182/20230160511.

Abstract

OBJECTIVE

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 fertilization-embryo transfer (IVF-ET) pregnancy.

METHODS

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.

RESULTS

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 ( <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 ( <0.05) and the expression levels of IL-10 and perforin were significantly decreased in the IVF-ET group ( <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 ( <0.05). There was no significant difference in the expression of IFN-γ, TNF-α, granzyme B, and granulysin, the cellular function indicators ( >0.05).

CONCLUSION

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.

摘要

目的

探讨体外受精 - 胚胎移植(IVF - ET)妊娠中,蜕膜中两种免疫细胞,即蜕膜自然杀伤(NK)细胞和γδT细胞在母胎界面的特征性功能变化。

方法

分别从12例自然妊娠(NP)妇女和32例IVF - ET妊娠妇女中采集蜕膜样本,她们分别被纳入NP组和IVF - ET组。然后将部分蜕膜样本进行石蜡包埋用于苏木精 - 伊红(HE)染色和免疫荧光染色,其余样本进行消化处理,并用聚蔗糖通过梯度离心法分离蜕膜免疫细胞(DICs)。采用流式细胞术测定NP组和IVF - ET组中蜕膜NK细胞和γδT细胞的细胞计数及其表面活化标志物CD69和NKG2D的表达水平。此外,还检测了细胞内细胞因子干扰素 -γ(IFN -γ)、肿瘤坏死因子 -α(TNF -α)、白细胞介素 -17A(IL -17A)和白细胞介素 -10(IL -10)以及溶细胞颗粒颗粒酶B、穿孔素和颗粒溶素的表达水平。对相关免疫指标的特征性变化进行比较和分析。

结果

组织标本的HE染色显示观察到了典型的蜕膜结构,且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)细胞的百分比显著低于NP组(<0.05)。对DICs进行流式细胞术分析显示,与NP组相比,IVF - ET组dNK细胞的百分比降低(<0.05),且IVF - ET组中IL -10和穿孔素的表达水平显著降低(<0.05)。然而,两组之间蜕膜γδT(dγδT)细胞计数无显著差异。IVF - ET组中IL -10、IL -17A和穿孔素的表达下调(<0.05)。细胞功能指标IFN -γ、TNF -α、颗粒酶B和颗粒溶素的表达无显著差异(>0.05)。

结论

IVF - ET妊娠妇女的dNK细胞计数以及dNK和dγδT细胞的一些细胞内细胞因子分泌在一定程度上降低,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研究。

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